Journal articles on the topic 'Blunted crossings'

The tragic story of Melquiades Estrada in Tommy Lee Jone’s prize-winning 2005 film could easily be one of the case histories in Oscar Martinez’s already classic Border People (1994), which brings togetherpersonal narratives that deal with cross-border migration, transnational interaction, irregular labor, ethnic confrontation and border control. Born and raised on the Texan-Mexican border, Jones is not unfamiliar with these dynamics taking place in border regions, which prove to be a unique human environment deeply marked by transnational processes and, yet, also signs of resistance on both sides to fully embrace or reject the other culture. The Three Burials is a serious attempt at incarnating the spirit of the place, documenting accurately its mixed culture, and describing the pain that most of its inhabitants suffer from. Several reviewers have rightly argued that Jones’ film, like Unforgiven (1992) and Lone Star (1996), “offers another twist on the Western genre, breaking conventions and proving that there is vast unexplored territory within the traditional gun-slinging setting of the frontier.” Elements such as the laconic use of language, the central role played by landscape or the paramount importance of violence and death are clearly reminiscent of a genre whose main conventions are still being fruitfully used for new purposes. Nevertheless, it would be an unpardonable critical blunder to think of Jones’ film as a mere continuation of a tradition that was mainly characterized by its excessive Manichaeism and its conviction that justice will be eventually recovered. This article argues that the message that Jones eventually sends to the audience is one full of ambivalence and ambiguity, and if some degree of justice is established at the end of the film, as Roger Ebert (2006) has noted, it is a ‘poetic justice’ more than a literal one.

Aim. To define principles of coronary wire choice during coronary artery chronic total occlusion (CTO) recanalization by antegrade approach. Material and methods. From 2009 to 2013 the attempt of coronary artery CTO recanalization by antegrade approach was undertaken for 217 patients. Depending on success of CTO crossing by coronary wire patients were divided into 2 groups: group 1 (n=164) - successful wire crossing, group 2 (n=53) -unsuccessful attempt of CTO recanalization by coronary wire.Results. In the group 2 there were more frequent CTO localization in the right coronary artery (p<0,05 as compared to group 1), presence of blunt stump, higher frequency of side branches in the zone of proximal CTO cup, and large CTO length (p<0,001 as compared to group 1). CTO recanalization in 47% cases begun from soft wire with polymeric coating and tapered tip, successful recanalization by these wire type was achieved in 49,02% patients. Using non-tapered tip intermediate stiffness wire as the first choice wire was associated with the failure of recanalization (p=0,0440 in intergroup comparison). At impossibility of CTO recanalization by soft wire with polymeric coating and tapered tip used as a first choice, it replacement on non-tapered tip intermediate stiffness wire allowed to achieve successful recanalization in 36,67% patients. In the cases of this strategy uneffectiveness using stiff wire with polymeric coating and tapered tip as a third choice allowed additionally to attain successful recanalization in 60% cases. Replacement of the soft wire with polymeric coating and tapered tip on stiff wire with polymeric coating and tapered tip, executable at presence of meaningful rigidity in the proximal CTO cup zone associated with the achievement the successful recanalization in 60% cases.Conclusion. On the basis of the obtained data it was created the conception of step-up wire stiffness increase during the CTO recanalization by antegrade approach.

The jugular foramen and the hypoglossal canal are both apertures located at the base of the skull. Multiple lower cranial nerve palsies tend to occur with injuries to these structures. The pattern of injuries tend to correlate with the combination of nerves damaged. Case Report: A 28-year-old male was involved in an AVP injury while crossing the highway. Exam showed a GCS of 15 AAOx3, with dysphagia, tongue deviation to the right, uvula deviation to the left and a depressed palate. Initial imaging showed B/L frontal traumatic Sub-Arachnoid Hemorrhages (tSAH), Left Frontal Epidural Hematoma and a Basilar Skull Fracture. On second look by a trained Neuroradiologist c At 3 month follow up, patient’s tongue normalized to midline and his dysphagia resolved. Discussion: Collette-Sicard syndrome is a rare condition/syndrome characterized by unilateral palsy of CN: IX, X, XII. This condition has been rarely described as a consequence of blunt head trauma. In most cases, the condition is self-limiting with patients regaining most to all of their neurological functions within 6 months. Nerve traction injuries and soft tissue edema compressing the cranial nerves are the leading two hypothesis. In conclusion, injuries with focal neurological deficits which were not apparent on initial imaging should be reviewed by relevant experts with concomitant knowledge of the patient’s history.

A “second liberation” swept the African continent beginning in 1989. In many places, multiparty elections and a measured optimism gained ground. Yet during the 1990s, the spirit of moderation and tolerance typical of the early independence movements began to fray. The recent armed conflicts of Central and West Africa and the columns of refugees crossing borders have served as a blunt reminder of the fragility of many of the continent’s democratic experiments.In this new era, law plays a central, visible, yet delicate role in many peace settlements and democratic transitions, from South Africa to Ghana. Africa’s courts have been challenged to provide the kinds of basic dispute resolution that lie at the core of what it means to be a “government.” At the same time, Africanjudges are mindful of Learned Hand’s caution in The Spirit of Liberty, taped above a secretary’s desk in Uganda. “Liberty lies in the hearts of men and women,” Hand wrote. “[W]hen it dies there, no constitution, no law, no court can save it; no constitution, no law, no court can even do much to help it.” The success of a postconflict transition will depend, in part, on the role of courts in sustaining a spirit of liberty and tolerance in their societies.

Abstract Most breast cancer deaths result from the development of metastases. The complex tumor microenvironment provides signals that can instruct both breast cancer cells to invade and tumor blood vessels to be leaky, hence supporting metastasis. We identified the receptor tyrosine kinase (RTK) AXL to be essential for metastasis. Genetic ablation of Axl in mouse models of HER2+ breast cancer, either globally or specifically in mammary epithelial cells, blunts metastasis but not primary tumor growth. We found that Axl expressed in tumor cells contributes to the remodeling of the tumor microenvironment, including immune cell recruitment and abnormal blood vessels. Hence, AXL in epithelial cells promotes cell invasion and shapes a pro-metastatic microenvironment that would be poorly responding to treatments such as immunotherapy. While AXL is known to be expressed on endothelial cells, its endothelial function has yet to be clearly defined. This project aims to address the hypothesis that AXL expressed on endothelial cells promotes processes that lead to abnormal blood vessel formation to promote metastasis. To address this hypothesis, we first studied the specific intracellular interactions between two RTKs, AXL and the known pro-angiogenic receptor VEGFR. Our preliminary analyses indicates that GAS6 can not only induce the phosphorylation of the permeability marker eNOS on its own but can also potentiate the VEGF’s response. The index of linearity of the endothelial tight junctions upon GAS6 stimulation show increased permeability of the endothelial cells. To further characterize the signaling pathways controlled by these receptors, we plan to perform a phosphoproteomics screen. We will focus on studying the AXL+VEGFR phosphoproteome and define the signaling pathways that impact vascular permeability. The most interesting candidate(s) will be studied in the context of mouse tumor models. In parallel, we are generating a conditional deletion of Axl in endothelial cells by crossing Axlfx/fx mice with Pdgfb:iCreER animals. With this, we will study the specific endothelial effects of Axl in vivo, specifically with vessel permeability and retinal angiogenesis assays. Multiple anti-angiogenic therapies for cancer failed in the clinic highlighting the need for new strategies to target endothelial cells in cancer. This project will provide essential information on the role of endothelial specific AXL, in the context of the HER2 breast cancer, and of its downstream effectors. With more specific targets, anti-angiogenic therapies could decrease their off-target effects and increase their efficacy in the clinic. This project has the potential to uncover therapeutic targets that could reduce the metastatic burden in breast cancer patients, and therefore better the prognostic for the HER2 cancer subtype. Citation Format: Andréane Lalonde, Jean-François Côté. The role of endothelial-specific AXL and its associated signaling pathways in the tumor microenvironment [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P6-12-05.

Traumatic optic neuropathy (TON) has been regarded a vision-threatening condition caused by either ocular or blunt/penetrating head trauma, which is characterized by direct or indirect TON. Injury happens during sports, vehicle accidents and mainly in military war and combat exposure. Earlier, we have demonstrated that remote ischemic post-conditioning (RIC) therapy is protective in TON, and here we report that AMPKα1 activation is crucial. AMPKα1 is the catalytic subunit of the heterotrimeric enzyme AMPK, the master regulator of cellular energetics and metabolism. The α1 isoform predominates in immune cells including macrophages (Mφs). Myeloid-specific AMPKα1 KO mice were generated by crossing AMPKα1Flox/Flox and LysMcre to carry out the study. We induced TON in mice by using a controlled impact system. Mice (mixed sex) were randomized in six experimental groups for Sham (mock); Sham (RIC); AMPKα1F/F (TON); AMPKα1F/F (TON+RIC); AMPKα1F/F LysMCre (TON); AMPKα1F/F LysMCre (TON+RIC). RIC therapy was given every day (5–7 days following TON). Data were generated by using Western blotting (pAMPKα1, ICAM1, Brn3 and GAP43), immunofluorescence (pAMPKα1, cd11b, TMEM119 and ICAM1), flow cytometry (CD11b, F4/80, CD68, CD206, IL-10 and LY6G), ELISA (TNF-α and IL-10) and transmission electron microscopy (TEM, for demyelination and axonal degeneration), and retinal oxygenation was measured by a Unisense sensor system. First, we observed retinal morphology with funduscopic images and found TON has vascular inflammation. H&E staining data suggested that TON increased retinal inflammation and RIC attenuates retinal ganglion cell death. Immunofluorescence and Western blot data showed increased microglial activation and decreased retinal ganglion cell (RGCs) marker Brn3 and axonal regeneration marker GAP43 expression in the TON [AMPKα1F/F] vs. Sham group, but TON+RIC [AMPKα1F/F] attenuated the expression level of these markers. Interestingly, higher microglia activation was observed in the myeloid AMPKα1F/F KO group following TON, and RIC therapy did not attenuate microglial expression. Flow cytometry, ELISA and retinal tissue oxygen data revealed that RIC therapy significantly reduced the pro-inflammatory signaling markers, increased anti-inflammatory macrophage polarization and improved oxygen level in the TON+RIC [AMPKα1F/F] group; however, RIC therapy did not reduce inflammatory signaling activation in the myeloid AMPKα1 KO mice. The transmission electron microscopy (TEM) data of the optic nerve showed increased demyelination and axonal degeneration in the TON [AMPKα1F/F] group, and RIC improved the myelination process in TON [AMPKα1F/F], but RIC had no significant effect in the AMPKα1 KO mice. The myeloid AMPKα1c deletion attenuated RIC induced anti-inflammatory macrophage polarization, and that suggests a molecular link between RIC and immune activation. Overall, these data suggest that RIC therapy provided protection against inflammation and neurodegeneration via myeloid AMPKα1 activation, but the deletion of myeloid AMPKα1 is not protective in TON. Further investigation of RIC and AMPKα1 signaling is warranted in TON.

Цель. Разработать модель прогнозирования успешности выполнения антеградной реканализации хронических окклюзий коронарных артерий коронарным проводником.Материалы и методы. Исследование являлось ретроспективным, одноцентровым и включало данные 391 пациента, которому в 2009–2018 годах была предпринята попытка реканализации хронических тотальных окклюзий (ХТО) коронарных артерий антеградным доступом. Все пациенты случайным образом были разделены на обучающую и экзаменационную выборки в соотношении 2:1. Построение шкалы прогнозирования включало определение предикторов успеха/неуспеха процедуры, вычисления баллов для каждого из них, с последующим суммированием всех полученных баллов для каждого поражения.Результаты и обсуждение. Успешное проведение проводника было выполнено в 73,7% случаев. Построенная многофакторная модель имела высокую статистическую значимость (c2=25,77, р<0,001). Важными предикторами, связанными с успешным антеградным проведением коронарного проводника, являлись 4 показателя: возраст поражения <12 месяцев (плюс 1 балл), плоская/неопределенная форма культи окклюзии (минус 2 балла), протяженность ХТО≥20 мм (минус 2 балла), выраженная извитость сосуда в зоне поражения (минус 1 балл). На основании суммарного балльного счета выделено 4 класса сложности ХТО для проведения антеградной реканализации коронарным проводником. Построенная модель показала хорошую дискриминационную способность (площадь под ROC-кривой 0,804 в обучающей выборке и 0,817 в экзаменационной выборке). Хорошая точность вероятностного прогнозирования полученной модели подтверждается значением показателя Brier, составившим 0,168.Выводы. Разработанная система дооперационного прогнозирования успешности антеградного проведения коронарного проводника через зону ХТО коронарной артерии позволяетпровести дифференцированный отбор пациентов для выполнения чрескожных коронарных вмешательств, что будет способствовать оптимизации качества оказания медицинской помощи данным пациентам. Purpose. To develop a predicting model of successful antegrade recanalization of chronic total occlusions of coronary arteries (CTO) with the coronary wire.Materials and methods. The study was retrospective, single-center. It included the data from 391 patients, who had attempts of antegrade CTO recanalization during 2009–2018. All patients were randomly divided into derivation and validation cohort (2:1 sampling ratio). The construction of the prognostic scale included determination of the predictors of success/failure of the procedure, calculating points for each of them, followed by summing up of all obtained points for each lesion. Results and discussion. Successful CTO crossing with coronary wire was performed in 73.7% of cases. The constructed multivariate model had high statistical significance (c2=25.77, p<0.001). Important predictors associated with successful antegrade recanalization with coronary wire are the lesion age <12 months (plus 1 point), blunt/ambiguous CTO stump (minus 2 points), occlusion length ≥20 mm (minus 2 points), severe tortuosity in the CTO zone (minus 1 point). On the base of the total score, 4 CTO difficulty classes were allocated for antegrade recanalization with a coronary wire. The model showed good discrimination ability (the area under the ROC curve is 0.804 in the derivation group and 0.817 in the validation group). The good accuracy of the probabilistic forecasting model is confirmed by the Brier score (0.168).Conclusions. The developed system of preoperative prediction of the success of antegrade CTO recanalization with coronary wire lets to differentiate the selection of patients for PCIs, which will optimize the quality of patient’s medical care.

Цель. Изучить особенности проведения реканализации хронических окклюзионных поражений коронарных артерий ретроградным доступом. Материалы и методы. В исследование включено 45 пациентов, которым в 20102019 гг. была предпринята попытка реканализации хронических тотальных окклюзий (ХТО) коронарных артерий ретроградным доступом. В зависимости от успеха выполненного вмешательства пациенты были разделены на 2 группы: группа 1 успешная реканализация (n26), группа 2 (n19) неуспешная реканализация. Результаты и обсуждение. Для пациентов обеих групп характерным являлось преобладание ХТО возрастом более 12 месяцев, доминирование плоской формы культи окклюзии, частое отхождение боковых ветвей в зоне проксимальной капсулы, редкая встречаемость извитости и кальциноза в зоне поражения. Успешное проведение инструментария по коллатералям было выполнено у 100 лиц группы 1 и 42,1 группы 2 (p0,001) наибольшая эффективность была достигнута при использовании мягких проводников с гидрофильным покрытием, без зауженного кончика (в 66,7 случаев), а также мягких проводников с полимерным и гидрофильным покрытием, без зауженного кончика (в 66,7). Ретроградная реканализация в группе 1 в 73,1 случаев была успешно проведена транслюминальным ретроградным способом, в 15,4 способом CART-M, в 11,5 reverseCART. Наибольший успех прохождения сквозь толщу ХТО был отмечен при использовании мягких (в 75) и жестких (в 44,4) проводников с зауженным кончиком. При невозможности выполнения ретроградной реканализации у пациентов группы 2 в 78,9 случаев проводилось изменение стратегии вмешательства на изолированную антеградную, что позволило достигнуть положительного эффекта у 20 обозначенных лиц. Заключение. Полученные данные позволяют детализировать технические аспекты реканализации ХТО коронарных артерий ретроградным доступом, что является актуальным для практического здравоохранения. Purpose. To study the peculiarities of recanalization of chronic occlusions of coronary arteries with retrograde approach Materials and methods. From 2010 to 2019, the attempt of coronary artery CTO recanalization with retrograde approach was made in 45 patients. Depending on success of CTO recanalization, patients were divided into 2 groups: group 1 (n26) successful procedure, group 2 (n19) unsuccessful procedure. Results and discussion. Patients of both groups were characterized by the predominance of CTO age 12 months, blunt stump, high rate of side branches in the CTO proximal cap area, and rare occurrence of tortuosity and calcification in the lesion. Successful delivery of instruments on collaterals was performed in 100 of persons of the group 1 and 42.1 of persons in the group 2 (p 0.001). The greatest efficiency was achieved in using soft wires with a hydrophilic coating and non-tapered tip (in 66.7 of cases), as well as soft wires with polymer and hydrophilic coating without tapered tip (in 66.7). Retrograde recanalization in the group 1 in 73.1 of cases was successfully performed with the transluminal retrograde CTO crossing, in 15.4 with the CART-M method, and in 11.5 with reverseCART. The greatest recanalization success was achieved after using soft (in 75) and hard (in 44.4) wires with the tapered tip. We changed the initial PCI strategy in 78.9 of cases of the group 2 to the isolated antegrade recanalization, which let to achieve positive result in 20 of patients. Conclusion. The obtained data let to detail the technical aspects of recanalization of CTO of coronary arteries with the retrograde approach, which is relevant for clinical practice.

Abstract Reference/Citation: Farion K, Osmond MH, Hartling L, et al. Tissue adhesives for traumatic lacerations in children and adults. Cochrane Database Syst Rev. 2001(4);CD003326. Clinical Question: What is the clinical evidence base for tissue adhesives in the management of simple traumatic lacerations? Data Sources: Studies were identified by searches of the following databases: Cochrane Wounds Group Specialized Trials Register (September 2003), Cochrane Central Register of Controlled Trials (CENTRAL) (CDROM 2003, issue 3), MEDLINE (1966 to September 2003, week 1), EMBASE (1988 to 2003, week 36), Web of Science Science Citation Index (1975 to September 13, 2003) and various clinical trials registers (September 2003). Investigators and product manufacturers were contacted to identify additional eligible studies. The search terms included wounds and injuries, laceration, face injury, nose injury, tissue adhesives, and acrylates. Study Selection: Each study fulfilled the following criteria: (1) The study was a randomized controlled trial that compared tissue adhesives with standard wound closure (SWC) (sutures, staples, adhesive strips) or tissue adhesive with tissue adhesive. (2) The wounds were acute, linear lacerations less than 12 hours old, resulting from blunt or sharp trauma. (3) The wound length, width, and depth allowed for approximation of the edges with minimal tension after deep sutures were placed, if required. Studies were included with no language or publication status restriction, with participants of any age recruited in an emergency department, outpatient clinic, walk-in clinic, or other primary care setting. Studies were excluded if the wounds were stellate lacerations, puncture wounds, mammalian bites, infected, heavily contaminated or devitalized, crossing joints or mucocutaneous junctions, in hair-bearing areas, or in patients with keloid formation or chronic illness. Data Extraction: The characteristics of the study and participants, interventions, outcome measures, and findings were extracted by one author and verified by a second using a standard form. The primary measure was cosmetic outcome. Secondary measures were pain with the procedure, time to complete the procedure, and complications (erythema, infection, discharge, need for delayed closure, and dehiscence). Studies were divided into 2 groups as follows: group 1, comparisons among tissue adhesives with SWC, and group 2, comparisons among different tissue adhesives. All eligible studies were assessed for methodologic quality independently by 2 investigators using the Jadad Scale, which evaluates randomization, double blinding, withdrawals, and dropouts and is scored on a 5-point (maximum) scale. The data from the tissue adhesive and SWC studies were pooled and analyzed with a random-effects model. The I2 statistic was used to determine heterogeneity among the studies. χ2 analysis was performed to compare participant age, wound location, and type of tissue adhesive among the studies. The data from the studies comparing tissue adhesives were pooled and analyzed using a fixed-effects model. Main Results: The search criteria identified 39 eligible studies, of which 11 met the inclusion criteria. In 10 studies, a tissue adhesive was compared with SWC. Five groups used butylcyanoacrylate, and 5 used octylcyanoacrylate. For SWC, 6 groups used sutures, 2 used adhesive strips, and 2 used a combination of methods, although most used sutures. Six studies were limited to pediatric patients and 2 to adult patients; 2 included patients of any age. Wounds were limited to facial lacerations in 2 pediatric studies and 1 group with patients of any age. Lacerations requiring deep sutures were excluded in 4 studies. One group compared tissue adhesives (butylcyanoacrylate and octylcyanoacrylate) among pediatric patients with facial lacerations not requiring deep sutures. In the 11 included studies, authors of 9 randomized and evaluated 1 laceration per patient, whereas 2 groups included patients with more than 1 laceration. In 1 group, each laceration was independently randomized and evaluated, and the other group randomized the patient and assigned all lacerations to a treatment group (tissue adhesive with SWC or tissue adhesive with tissue adhesive). The sample sizes ranged between 60 and 163 lacerations, and all 11 studies were performed in emergency departments. The primary measure in all included studies was cosmetic outcome. The majority of groups used the Cosmetic Visual Analogue Scale, the Wound Evaluation Score, or a combination of these measures. Three groups measured cosmetic outcome with nonvalidated scoring systems. Assessment time periods were grouped and reported at (1) 5 to 14 days, (2) 1 to 3 months, and (3) 9 to 12 months after wound closure. Secondary outcomes were pain (as noted on visual analogue scale) and time to complete the procedure (as mean number of minutes). The 11 studies scored from 1 to 3 on the Jadad Scale. Adequate allocation concealment was reported in only 1 group. Examining cosmetic outcome, 8 groups (565 lacerations) used the Cosmetic Visual Analogue Scale to compare tissue adhesives and SWC. The authors reported no significant differences in scores at the time periods of 5 to 14 days, 1 to 3 months, and 9 to 12 months. A subgroup analysis showed a significant (P = .005) superiority of butylcyanoacrylate over SWC at 1 to 3 months. Using the Wound Evaluation Score, 4 studies (364 lacerations) compared tissue adhesives with SWC. No significant differences in cosmetic scores were found at 5 to 14 days, 1 to 3 months, or 9 to 12 months. One group (83 lacerations) compared butylcyanoacrylate with octylcyanoacrylate and reported no significant differences in cosmetic scores using the Cosmetic Visual Analogue Scale at 1 to 3 months and the Wound Evaluation Score at 5 to 14 days and 1 to 3 months. Examining secondary outcomes, 6 groups (570 lacerations) compared tissue adhesives with SWC using the visual analogue scale for pain. Scores reported by parents, patients, physicians, and nurses significantly favored tissue adhesives. In 6 studies (584 lacerations), tissue adhesives were significantly favored over SWC in time to complete the procedure. For complication outcomes, 8 groups (727 lacerations) demonstrated significantly fewer incidences of erythema and an increased risk of dehiscence with tissue adhesives compared with SWC. No significant differences were shown for infection, delayed closure, or discharge. Among 83 lacerations, 1 group compared butylcyanoacrylate with octylcyanoacrylate and reported no significant differences in combined patient-reported and parent-reported visual analogue pain scores, time to complete the procedure, dehiscence, or infection. Conclusions: This review provides evidence that tissue adhesives are an option to SWC (sutures, staples, adhesive strips) for the management of simple traumatic lacerations. Overall, no significant differences were found in cosmetic scores at the reported assessment periods between tissue adhesives and SWC. At 1 to 3 months, a subgroup analysis significantly favored butylcyanoacrylate over SWC. Tissue adhesives significantly lowered the time to complete the procedure, levels of pain, and rate of erythema. However, the data revealed a significant increase in the rate of dehiscence with the use of tissue adhesives when compared with SWC. The low methodologic quality of the evidence should be considered in the interpretation of the findings.

Pompe disease (PD) is caused by a loss of function of the enzyme acid-α-glucosidase (GAA) leading to glycogen accumulation, neuromuscular dysfunction, and breathing failure. Here, we characterized longitudinal changes in breathing of GAA null ( Gaa−/−) rats on a breath-by-breath basis using a novel respiratory event detection algorithm. Adult Pompe (n = 5) and Sprague Dawley (n = 5) rats were implanted with chronic diaphragm EMG electrodes. Once a month from 4-10 months of age, full body plethysmography with concurrent EMG was used to record respiratory waveforms under room air and hypoxia. Waveforms were analyzed by the Adjustable Baselines Respiratory Analysis Program (ABRAP) algorithm. An adaptive threshold identified “respiratory events” defined as crossings of a threshold set between the recent maxima and minima of the waveform. After event identification, a range of waveform characteristics are calculated and tagged to that event. Averages of each characteristic were taken for all events and then analyzed based on respiratory rate in breaths/minute (bpm): low (<120 bpm, ‘quiet breathing’), medium (between 120 and 240 bpm) and high (>240 bpm, ‘high frequency sniffng’). In room air, Pompe rats had a lower breathing rate than wildtype rats by month 10 (173 vs 285 bpm, standard error of the difference (SE of diff.) 37, p=0.03). This decrease in frequency was driven by the high rate events as there were no differences in the mean rate of low rate events at month 10 (80 vs 83 bpm, SE of diff. 3) while the frequency of high rate events was significantly lower by 9 months (mo 9: 393 vs 440 bpm, SE of diff. 16, p=0.04, mo 10: 383 vs 440 bpm, SE of diff. 17, p=0.008). Not only was the breathing frequency of high-rate events impaired in Pompe rats, but there was a trend towards decrease in the proportion of events falling in the high rate category indicating less time spent in high rate breathing behaviors (mo 4: 52 vs 59%, SE of diff. 6, mo 10: 25 vs 52%, SE of diff. 9) with a concomitant increase in the proportion of low frequency events. Additionally, there was a main effect of genotype on the latency between diaphragm activation and onset of inspiratory flow during hypoxia with Pompe rats having a longer latency across all months and frequency bands. This difference was most pronounced in the medium (44 vs 29 ms, SE of diff. 4 ms, p=0.003) frequency events. Active expiration was impaired in Pompe animals with peak post-event airflow (maximum positive pressure reached before the next inspiratory effort) blunted across all months. This effect was pronounced in the high frequency events (0.16 mL vs 0.24 mL, SE of diff. 0.01, p=0.0001). These deficits in high frequency events are consistent with progressive neuromuscular diaphragm weakness. We conclude that comprehensive analyses of all respiratory events over extended recording periods enables detection of altered breathing behaviors that may be missed when assessing only short periods of quiet breathing. In turn, this may help uncover a respiratory signature of disease progression in Pompe disease or other neuromuscular conditions. 2R01HD052682-11A1 (DDF, BJB), T32HL134621 (AM), R01HL153102 (ED), SPARC OT2OD023854 (ED), Craig H. Neilsen Pilot Grant (ED). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

AbstractIn percutaneous coronary intervention (PCI) to the culprit lesion of acute myocardial infarction (AMI), unsuccessful guidewire crossing causes immediate poor outcomes. It is important to determine the factors associated with unsuccessful guidewire crossing in AMI lesions. The purpose of this study was to find factors associated with difficulty in crossing the culprit lesion of AMI. We defined the difficult group when the guidewire used to cross the culprit lesion was a polymer jacket type guidewire or a stiff guidewire. We included 937 patients, and divided those into the non-difficult group (n = 876) and the difficult group (n = 61). Proximal reference diameter was significantly smaller in the difficult group than in the non-difficult group (p < 0.001), and degree of calcification was severer in the difficult group than in the non-difficult group (p < 0.001). In the multivariate stepwise logistic regression analysis, proximal reference diameter [odds ratio (OR) 0.313, 95% confidence interval (CI) 0.185–0.529, p < 0.001)], previous PCI (OR 3.065, 95% CI 1.612–5.830, p = 0.001), moderate-severe calcification (OR 4.322, 95% CI 2.354–7.935, p < 0.001), blunt type obstruction (OR 12.646, 95% CI 6.805–23.503, p < 0.001), and the presence of collateral to the culprit lesion (OR 2.110, 95% CI 1.145–3.888, p = 0.017) were significantly associated with difficulty in crossing the culprit lesion. In conclusion, previous PCI, calcification, blunt type obstruction, and the presence of collateral were associated with difficulty in crossing the culprit lesion, whereas proximal reference diameter was inversely associated with difficulty. Our study provides a reference to recognize the difficulty in crossing the culprit lesions of AMI for PCI operators, especially junior operators.

Abstract Background RHAMM is a multifunctional protein that is upregulated in breast tumors, and the presence of strongly RHAMM+ve cancer cell subsets associates with elevated risk of peripheral metastasis. Experimentally, RHAMM impacts cell cycle progression and cell migration. However, the RHAMM functions that contribute to breast cancer metastasis are poorly understood. Methods We interrogated the metastatic functions of RHAMM using a loss-of-function approach by crossing the MMTV-PyMT mouse model of breast cancer susceptibility with Rhamm−/− mice. In vitro analyses of known RHAMM functions were performed using primary tumor cell cultures and MMTV-PyMT cell lines. Somatic mutations were identified using a mouse genotyping array. RNA-seq was performed to identify transcriptome changes resulting from Rhamm-loss, and SiRNA and CRISPR/Cas9 gene editing was used to establish cause and effect of survival mechanisms in vitro. Results Rhamm-loss does not alter initiation or growth of MMTV-PyMT-induced primary tumors but unexpectedly increases lung metastasis. Increased metastatic propensity with Rhamm-loss is not associated with obvious alterations in proliferation, epithelial plasticity, migration, invasion or genomic stability. SNV analyses identify positive selection of Rhamm−/− primary tumor clones that are enriched in lung metastases. Rhamm−/− tumor clones are characterized by an increased ability to survive with ROS-mediated DNA damage, which associates with blunted expression of interferon pathway and target genes, particularly those implicated in DNA damage-resistance. Mechanistic analyses show that ablating RHAMM expression in breast tumor cells by siRNA knockdown or CRISPR-Cas9 gene editing blunts interferon signaling activation by STING agonists and reduces STING agonist-induced apoptosis. The metastasis-specific effect of RHAMM expression-loss is linked to microenvironmental factors unique to tumor-bearing lung tissue, notably high ROS and TGFB levels. These factors promote STING-induced apoptosis of RHAMM+ve tumor cells to a significantly greater extent than RHAMM−ve comparators. As predicted by these results, colony size of Wildtype lung metastases is inversely related to RHAMM expression. Conclusion RHAMM expression-loss blunts STING-IFN signaling, which offers growth advantages under specific microenvironmental conditions of lung tissue. These results provide mechanistic insight into factors controlling clonal survival/expansion of metastatic colonies and has translational potential for RHAMM expression as a marker of sensitivity to interferon therapy.

Abstract Aims NADPH oxidase (NOX) 1 but not NOX4-dependent oxidative stress plays a role in diabetic vascular disease, including atherosclerosis. Endothelin (ET)-1 has been implicated in diabetes-induced vascular complications. We showed that crossing mice overexpressing human ET-1 selectively in endothelium (eET-1) with apolipoprotein E knockout (Apoe−/−) mice enhanced high-fat diet-induced atherosclerosis in part by increasing oxidative stress. We tested the hypothesis that ET-1 overexpression in the endothelium would worsen atherosclerosis in type 1 diabetes through a mechanism involving NOX1 but not NOX4. Methods and results Six-week-old male Apoe−/− and eET-1/Apoe−/− mice with or without Nox1 (Nox1−/y) or Nox4 knockout (Nox4−/−) were injected intraperitoneally with either vehicle or streptozotocin (55 mg/kg/day) for 5 days to induce type 1 diabetes and were studied 14 weeks later. ET-1 overexpression increased 2.5-fold and five-fold the atherosclerotic lesion area in the aortic sinus and arch of diabetic Apoe−/− mice, respectively. Deletion of Nox1 reduced aortic arch plaque size by 60%; in contrast, Nox4 knockout increased lesion size by 1.5-fold. ET-1 overexpression decreased aortic sinus and arch plaque alpha smooth muscle cell content by ∼35% and ∼50%, respectively, which was blunted by Nox1 but not Nox4 knockout. Reactive oxygen species production was increased two-fold in aortic arch perivascular fat of diabetic eET-1/Apoe−/− and eET-1/Apoe−/−/Nox4−/− mice but not eET-1/Apoe−/−/Nox1y/− mice. ET-1 overexpression enhanced monocyte/macrophage and CD3+ T-cell infiltration ∼2.7-fold in the aortic arch perivascular fat of diabetic Apoe−/− mice. Both Nox1 and Nox4 knockout blunted CD3+ T-cell infiltration whereas only Nox1 knockout prevented the monocyte/macrophage infiltration in diabetic eET-1/Apoe−/− mice. Conclusion Endothelium ET-1 overexpression enhances the progression of atherosclerosis in type 1 diabetes, perivascular oxidative stress, and inflammation through NOX1.

AbstractBackgroundIntraplaque delivery of contrast has been utilized during percutaneous coronary interventions (PCI) of chronic total occlusions (CTO) to delineate anatomy and to facilitate wire crossing. Its utility as a tool to accomplish primary crossing of CTOs has not been described or validated.AimsWe describe a new technique leveraging the diagnostic and therapeutic roles of intraplaque contrast injection to accomplish primary crossing of CTOs: HydroDynamic contrast Recanalization (HDR).MethodsHDR is an antegrade crossing method for coronary CTOs based on the synergistic use of contrast microinjections and polymer jacketed wires. We present a retrospective, first‐in‐man, case series utilizing HDR for CTO PCI in patients with favorable CTO anatomy (visible proximal segment and identifiable distal target). The primary outcome was procedural success. The secondary outcome was any procedural complications.ResultsA total of 43 patients with 45 CTOs underwent CTO PCI with HDR. Mean patient age was 64.3 ± 11 years. The mean Japanese CTO and PROGRESS CTO scores were 2.3 ± 0.7 and 1.8 ± 0.7, respectively. CTO complexity was high, with an ambiguous or blunt cap in 34 occlusions (76%); lesion length ≥ 20 mm in 27 occlusions (60%); and moderate/heavy calcification in 36 occlusions (80%). Procedural success using HDR was 100%. There were no complications.ConclusionsThis study shows the utility of HDR in CTO PCI. HDR appears to be a safe and promising new contrast‐based primary crossing technique in selected patients. This strategy warrants further evaluation in larger prospective studies.

Abstract Background Chronic Coronary Total Occlusion (CTO) is considered one of the most challenging obstructive coronary artery diseases that had been widely managed medically or referred for bypass surgeries due to their complex nature which remained an obstacle for the operators. Coronary CT- angiography was used to develop easy predictive tools for CTO percutaneous intervention. Objectives To evaluate the ability of the CCTA and CT-RECTOR score to predict time-efficient guidewire (GW) crossing through a chronic total occlusion. Patients and Methods The present study included 40 patients with CTO lesions. All were subjected to preoperative CTO assessment using CCTA and CCA before an attempt of PCI to a CTO in the period between January 2022 and May 2023. The study was held in Ain Shams University hospitals and National Heart Institute. Results We divided the patients into two groups according to GW crossing within 30 minutes. Group 1 included patients with successful GW crossing within 30 min, group 2 included patients with failed GW crossing within 30 min. We compared the two groups according to demographic data and we found that males are more common to have CTO lesions (87.5%) which were predominantly located in RCA (50% of total no. with 76.9% of them in failed group, p = 0.018). Our results showed that CTOs with occlusion length ≥ 20 mm or calcification increased in failed group, but was not statistically significant (84.6% in failed group (p = 0.161) and 46.3% in failed group (p = 0.432) respectively). In addition, there was a statistical significance between presence of blunt stump (76.9% in failed group vs 29.6% in successful group, p = 0.004), a previous failed CTO-PCI attempt (53.8% in failed group vs 14.8% in successful group p = 0.009), and using retrograde wiring techniques (p = 0.003) between the successful and failed groups. Indicating that these points were important predictors of procedural failure. We compared CTO characteristics between CCTA-derived CT-RECTOR score and CA-derived J-CTO score. with increased score points, the probability of PCI difficulty and GW crossing failure increased. CT-RECTOR score was found to have the better ability for prediction of GW crossing within 30 min and CTO-PCI difficulty (p = 0.028). Conclusion CT-RECTOR score was superior to J-CTO score in the assessment and prediction of GW crossing difficulty within 30 minutes and final procedural outcomes.

AimsChronic total occlusion (CTO) percutaneous coronary intervention (PCI) is characterized by a low success rate and an increase in complications. This study aimed to explore a new and simple classification method based on plaque composition to predict guidewire (GW) crossing within 30 min of CTO lesions.MethodsThis study consecutively enrolled individuals undergoing attempted PCI of CTO who underwent coronary computed tomographic angiography (CCTA) within 2 months. Lesions were divided into soft and hard CTO groups according to the necrotic core proportion.ResultsIn this study, 207 lesions were divided into soft (20.3%) and hard CTO (79.7%) groups according to a necrotic core percentage cutoff value of 72.7%. The rate of successful GW crossing within 30 min (57.6 vs. 85.7%, p = 0.004) and final success (73.3 vs. 95.2%, p = 0.001) were much lower in the hard CTO group. For patients with hard CTO, previous failed attempt, proximal side branch, bending > 45 degrees calcium ≥ 50% cross-sectional area (CSA), and distal reference diameter ≤ 2.5 mm were demonstrated to be associated with GW failure within 30 min. For patients with soft CTO, only blunt entry was proved to be an independent predictive factor of GW failure within 30 min.ConclusionsGrouping CTO lesions according to the proportion of necrotic core is reasonable and necessary in predicting GW crossing within 30 min. A soft CTO with a necrotic core is more likely to be recanalized compared with a hard CTO with fibrous and/or dense calcium. Different plaque types have variable predictive factors.

Abstract Objective This study aimed to establish a new scoring system that includes histological quantitative features derived from coronary computed tomographic angiography (CCTA) to predict the efficiency of chronic total occlusion percutaneous coronary intervention (CTO-PCI). Methods This study analyzed clinical, morphological, and histological characteristics of 207 CTO lesions in 201 patients (mean age 60.0 [52.0–65.0] years, 85% male), which were recruited from two centers. The primary endpoint was a guidewire successfully crossing the lesions within 30 m. The new predictive model was generated by factors that were determined by multivariate analysis. The CCTA plaque (CTAP) score that included a quantitative plaque characteristic was developed by assigning an appropriate integer score to each independent predictor, then summing all points. In addition, the CTAP score was compared with other predictive scores based on CCTA. Results The endpoint was achieved in 63% of the lesions. The independent predictors included previous CTO-PCI failure, the proximal blunt stump, proximal side branch, distal side branch, occluded segment bending > 45°, and high-density plaque volume (fibrous volume + calcified volume) ≥ 19.9 mm3. As the score increased from 0 to 5, the success rate of the guidewire crossing within 30 m decreased from 96 to 0%. Comparing the CTAP score with other predictive scores, the CTAP score showed the highest discriminant power (c-statistic = 0.81 versus 0.73–0.77, p value 0.02–0.07). The CTAP score showed similar results for procedural success. Conclusion The CTAP score efficiently predicted the guidewire crossing efficiency and procedural success. Key Points • An increase in high-density plaque volume (fibrous + dense calcium) was more probable to reduce the efficiency of crossing and lead to procedural failure. • The new prediction scoring system with the addition of the quantitative characteristics of plaques had an improved predictive ability compared with the traditional prediction scoring system.

Abstract Traumatic injury in children often involves traffic accidents, falls from height, bicycle accidents, abuse, and sports. We present our experience with a 6-year-old girl who bruised her abdomen by falling while running with a thermos bottle hanging diagonally across her neck and left shoulder, crossing in front of her body. After the injury, she was rushed to the hospital and plain abdominal computed tomography revealed free air, suggesting intestinal injury due to blunt trauma. She was transferred to our hospital, where abdominal examination revealed a bruise and redness at the umbilicus. There were signs of peritoneal irritation and guarding throughout the abdomen. Upright chest X-ray showed free air in the right subdiaphragmatic region. Laparoscopic inspection revealed one perforation and two serosal injuries. The damaged serosa was repaired, and the perforated intestine was resected and anastomosed. The patient was discharged on the 11th day following an uneventful postoperative course.

Central neurotensin signaling via neurotensin receptor-1 (NtsR1) modulates various aspects of physiology, including suppressing feeding and promoting locomotor activity that can support weight loss. However, it remains unclear when and where NtsR1 expression contributes to control of body weight vs. other effects. We previously showed that activating ventral tegmental area (VTA) dopamine (DA) neurons that express NtsR1 promotes weight loss. We therefore hypothesized that deleting NtsR1 from DA neurons would promote weight gain by increasing food intake and decreasing physical activity. In contrast, developmental deletion of NtsR1 from DA neurons (by crossing DATCre mice with NtsR1flox/flox mice) had no impact on the feeding or body weight of mice fed a chow diet, though it augmented locomotor activity. Developmental deletion of NtsR1 from DA neurons protected mice from diet-induced obesity, but not via altering feeding, physical activity, or energy expenditure. Given that NtsR1 may exert distinct roles within development vs. adulthood, we then examined the impact of adult-onset deletion of NtsR1 from VTA DA neurons. We injected adult NtsR1flox/flox mice in the VTA with adeno associated virus to Cre-dependently delete NtsR1 in the VTA (VTAR1Null mice) and compared them to mice with intact NtsR1 (Controls). Again, in contrast to our hypothesis, VTAR1Null mice gained less weight than Controls while on normal chow or high fat diets. Moreover, VTAR1Null mice exhibited blunted feeding after fasting, suggesting a role for NtsR1 in adult VTA DA neurons in coordinating energy need and intake. Altogether, these data suggest that intact expression of NtsR1 in DA neurons is necessary for appropriate regulation of body weight, but a lack of NtsR1 in the developing vs. adult DA system protects from weight gain via different mechanisms. These findings emphasize the need for temporal and site-specific resolution to fully understand the role of NtsR1 within the brain.

Background. Cardiac hypertrophy is characterized by transcriptional reprogramming of fetal gene expression, and histone modifiers are tightly linked to the regulation of those genes. We previously reported that activation of histone deacetylase (HDAC) 2, one of the class I HDACs, mediates hypertrophy. Here we suggest that disinhibiting of kruppel-like factor 4 (Klf4) by casein kinase-2α1 (CK2α1)-dependent phosphorylation of HDAC2 S394 develop the cardiac hypertrophy. Methods and Results. Hypertrophic stimuli phosphorylated Hdac2 S394, which was necessary for its enzymatic activation and thereby for the development of hypertrophic phenotypes. Transgenic mice overexpressing Hdac2-wild type exhibited cardiac hypertrophy, whereas those expressing phosphorylation-resistant Hdac2 S394A did not. Compared with that in age-matched normal human hearts, phosphorylation of Hdac2 S394 was dramatically increased in hypertrophic cardiomyopathy patients. Hypertrophy-induced phosphorylation of Hdac2 S394 and its enzymatic activity were completely blocked either by CK2-blockers or by CK2a1 siRNA. Hypertrophic stimuli led CK2α1 to be activated, and its chemical inhibitors blocked hypertrophy in both phenylephrine-treated cardiomyocytes and in isoproterenol-administered mice. However, by utilizing KLF4-binding element-disrupted Nppa promoter, treatment with either TBB or TBCA failed to reduce the mutant promoter activity. These results emphasized that CK2α1-induced hypertrophic events are dependent on both Hdac2 and KLF4. CK2α1-transgenic mice developed hypertrophy, which was attenuated by administration of trichostatin A, an HDAC inhibitor. Overexpression of CK2α1 caused hypertrophy in cardiomyocytes, whereas its chemical inhibitors as well as Hdac2 S394A blunted it. Hypertrophy in CK2α1-transgenic mice was exaggerated by crossing these mice with Hdac2-transgenic mice. By contrast, however, it was blocked when CK2α1-transgenic mice were crossed with Hdac2 S394A-transgenic mice. Conclusions. We have demonstrated a novel mechanism in the development of cardiac hypertrophy by which CK2 activates HDAC2 via phosphorylating HDAC2 S394 and consequence down-regulation of KLF4.

Macrophages play a key role in the pathogenesis of atherosclerosis. Metabolic programs powered by glucose or lipid enable macrophages to elicit pro- or anti-inflammatory responses, respectively. Although the chronic inflammatory feature of atherosclerosis has been well-established, the role of macrophage substrate metabolism in atherogenesis remains unclear. We previously demonstrated that macrophages with elevated GLUT1-mediated glucose metabolism have an increased inflammatory response. Therefore, we created a novel macrophage GLUT1-deficient murine model by crossing GLUT1 floxed to LysM-Cre mice. Recent work suggests that lack of GLUT1 reduces the pro-inflammatory response and the ability for GLUT1-/- macrophages to polarize to the classically activated state in vitro. Therefore, the objective of this study was to examine how macrophage metabolic reprogramming affects the development of atherosclerosis. We hypothesized that macrophages with restricted glucose metabolism due to lack of glucose transporter GLUT1 will have reduced pro-inflammatory activation during atherogenesis. We transplanted bone marrow from Glut1MΦfl/fl or Glut1MΦ-/- mice into Ldlr-/- mice and fed mice a Western diet for 12 weeks. Glut1MΦfl/fl Ldlr-/- or Glut1MΦ-/- Ldlr-/- chimeric mice did not exhibit significant differences in body weight, body composition, blood pressure, fasting blood glucose, or triacylglycerol and LDL and HDL cholesterol. Digital histology analysis of Oil Red O stained slides indicated that deleting macrophage GLUT1 did not affect total lesion area in aortic root; however, mice with blunted glucose metabolism displayed more and larger necrotic cores. Ongoing studies are investigating apoptosis and phagocytic capacity of macrophages with and without GLUT1 to elucidate the roles of macrophage glucose metabolism on the morphology of atherosclerotic lesion. In summary, we observed that mice lacking macrophage GLUT1 developed increased necrotic cores in lesions of the aorta root relative to mice with wild type macrophage GLUT1 which suggests that maintenance of atherosclerotic lesion stability may be regulated by glucose-dependent mechanisms.

Previously, a homodiploid goldfish-like fish (2n = 100; GF-L) was spontaneously generated by self-crossing a homodiploid red crucian carp-like fish (2n = 100; RCC-L), which was in turn produced via the distant hybridization of female koi carp (Cyprinus carpio haematopterus, KOC, 2n = 100) and male blunt snout bream (Megalobrama amblycephala, BSB, 2n = 48). The phenotypes and genotypes of RCC-L and GF-L differed from those of the parental species but were similar to diploid red crucian carp (2n = 100; RCC) and goldfish (2n = 100; GF), respectively. We sequenced the complete mitochondrial DNAs (mtDNAs) of the KOC, BSB, RCC-L, GF-L, and subsequent generations produced by self-crossing [the self-mating offspring of RCC-L (RCC-L-F2) to the self-mating offspring of RCC-L-F2 (RCC-L-F3) and the self-mating offspring of GF-L (GF-L-F2)]. Paternal mtDNA fragments were stably embedded in the mtDNAs of both lineages, forming chimeric DNA fragments. In addition to these chimeras, several nucleotide positions in the RCC-L and GF-L lineages differed from the parental bases, and were instead identical with RCC and GF, respectively. Moreover, RCC-L and GF-L mtDNA organization and nucleotide composition were more similar to those of RCC and GF, respectively, compared to parental mtDNA. Finally, phylogenetic analyses indicated that RCC-L and GF-L clustered with RCC and GF, not with the parental species. The molecular dating time shows that the divergence time of KOC and GF was about 21.26 Mya [95% highest posterior density (HPD): 24.41–16.67 Mya], which fell within the period of recent. The heritable chimeric DNA fragments and mutant loci identified in the mtDNA of the RCC-L and GF-L lineages provided important evidence that hybridizations might lead to changes in the mtDNA and the subsequent generation of new lineages. Our findings also demonstrated for the first time that the paternal mtDNA was transmitted into the mtDNA of homodiploid lineages (RCC-L and GF-L), which provided evidence that paternal DNA plays a role in inherited mtDNA. These evolutionary analyses in mtDNA suggest that GF might have diverged from RCC after RCC diverged from koi carp.

Background: Prevalence in Indonesian society in 2007 was 7.5%, with several factors causing traffic and crossing sharp/ blunt objects. In 2013 there was an increase in the prevalence of trauma to 8.2%, with the highest order of injuries being down 40.9%, motorcycle accidents (40.6%), trauma due to sharp/blunt objects 7.3%, other land transportation 7, 1% and 2.5% fallout. To equip ordinary students in carrying out the Log roll Skills to help victims with spinal trauma need to do log roll Skills training. In this regard, researchers intend to conduct research on "The Effect of Health Education with First Aid (Log Roll) Simulation Methods on Spinal Injuries in Tempuran Informal Education in Magelang". Methods: This research is experimental research with the PreTest-Post Test Group design approach. through hypothesis testing research. The population of this research was 50 people in the cloud (Ponpes Roudlotuttulab students). Statistical test on paired groups using the nonparametric Wilcoxon test. Result: Based on the Wilcoxon test results, it can be concluded that 50 respondents who log roll training can be drawn in the conclusion that the majority of students have a basic score of 0.001 which means there is a difference that takes place between before and after training. Conclusion: The suggestion put forward in this study is that education and log roll action training for lay people, especially students, continues to be held at other Islamic boarding schools to improve the skills of log roll action skills suspected in victims with spinal injuries.

Leptin acts in the brain to decrease food intake and promote energy expenditure by increasing sympathetic nerve activity (SNA) to thermogenic brown adipose tissue. Leptin also increases SNA to other beds including kidney with implications for obesity-induced hypertension. We previously demonstrated the importance the arcuate nucleus (Arc) of the hypothalamus in mediating leptin-induced increases in regional SNA, but the specific neuronal population within the Arc that mediates these responses is unknown. We hypothesized that agouti-related peptide (AgRP) and/or proopiomelanocortin (POMC) neurons of the Arc are critical for the increases in SNA in response to leptin. To test this, we generated mice lacking the leptin receptor specifically in AgRP or POMC neurons by crossing the LepR flox/flox mice with AgRP Cre or POMC Cre mice. Consistent with previous reports, both AgRP Cre/ LepR flox/flox mice and POMC Cre /LepR flox/flox mice have a slightly elevated body weight relative to littermate controls. Next, we used multifiber sympathetic nerve recording to assess the SNA effects of leptin. Intracerebroventricular (ICV) injection of leptin (2 μg) led to a comparable increase in renal SNA in control mice (210±93%) and AgRP Cre /LepR flox/flox mice (191±53%). AgRP Cre/ LepR flox/flox mice also displayed a normal lumbar SNA response to ICV leptin (384±86%) relative to controls (325±46%). In contrast, POMC Cre /LepR flox/flox mice exhibited a significantly reduced renal SNA response to ICV leptin (5±17%) as compared to controls (174±45%). Lumbar SNA response to leptin was also blunted in POMC Cre /LepR flox/flox mice (24±21%) as compared to controls (358±53%). Thus, deletion of the leptin receptor from POMC neurons, but not AgRP neurons, interferes with the ability of leptin to increase sympathetic traffic. These results demonstrate that the sympathetic nerve responses evoked by leptin emanate from leptin action on POMC neurons.

Abstract Aim Cardiovascular diseases (CVDs) are the leading cause of death globally, claiming an estimated 17.9 million lives each year. A better understanding of the underlying chronic inflammatory disease mechanisms in atherosclerosis might help finding novel treatments and reduce CVD deaths. In particular, the cannabinoid receptor CB1 has been implicated in atherosclerosis, while its cell-specific effects in this disease are not well understood. Methods Myeloid Cnr1 (CB1 encoding gene) knockout mice were generated on apolipoprotein E deficiency (Apoe-/-) background by crossing CB1flox mice with transgenic mice carrying Cre under control of the lysozyme M (LysM) promoter for selective expression in myeloid cells. Age - and sex-matched groups were analyzed at baseline, 4 and 16 weeks Western diet (0,15% cholesterol). For in vitro experiments, murine bone marrow-derived macrophages (BMDM) were used. Results Male mice with myeloid-specific Cnr1 deficiency on atherogenic background developed smaller lesions and necrotic cores compared to controls, while only minor effects in females were observed. Male Cnr1 deficient mice had reduced arterial monocyte recruitment and macrophage proliferation with less inflammatory phenotype. The sex-specific differences were reproducible in vitro in BMDMs and blunted by female hormone estradiol treatment. Array-based kinase activity profiling of BMDMs stimulated with a synthetic CB1 agonist revealed a CB1-dependent regulation of p53 and cyclin-dependent kinases. Transcriptomic profiling via bulk RNA-sequencing of CB1 agonist-stimulated BMDMs further unveiled chromatin modifications, mRNA processing and mitochondrial respiration among the key processes affected by CB1 signaling, which was supported by metabolic flux assays. Chronic administration of the peripherally-restricted CB1 antagonist JD5037 inhibited plaque progression and macrophage proliferation, again only in male mice. Conclusion Impaired CB1 signaling in macrophages is atheroprotective by limiting their arterial recruitment, proliferation and inflammatory reprogramming. The biological effects of macrophage CB1 signaling seem to be more pronounced in male mice. This highlights the need to consider the biological sex as an important variable in preclinical studies.

This study examined the hatchability performance of the offspring and some egg quality characteristics, which will be obtained from crossing Leghorn breed chickens and five different genotypes. The study's experiment was carried out in the XXXX . In the present study, which was designed to have one male and twelve females belonging to each genotype, a total of 200 eggs from each flock were examined. No adverse results were found in the incubation results of the crosses made with our local breed Denizli and Araucana, Brahma, and Cornish breeds, whose breeders increased locally. When egg quality characteristics were examined, the difference in egg weight between F1 genotypes was insignificant and ranged between 46.91-51.54 g on average. When the F2 generation was investigated, egg weight differed between genotype groups, and the average weights ranged between 57.6-67.14 g. In addition, the effect of genotype on strength, bark thickness, and point blunt up shell thickness values was significant. However, the effect on the Haugh Unit and yellow height were insignificant in the same generation. As a result, hybridizing genotypes with low yield performance with commercially important genotypes could provide a genotype for alternative production systems for future generations.

Abstract Introduction Implantation of a penile prosthesis in cases of corporeal fibrosis is a challenging procedure. Cavernotomes aid excavation of fibrous tissue in scarred corporal bodies. However, in many cases, the track established -with risk and difficulty- barely suffices for a narrow base inflatable or girth-9 malleable rods. The patient may have this exchanged with a full-size implant after several months, in another surgery. Shaeer’s Cavernotome is a corer, that cores out and grinds fibrous tissue in gentle rotatory movements rather than forceful forward thrusting. The process is therefore safer, and with lower risk of perforation. By choosing the maximum size of Shaeer’s cavernotome (6-13 Hegar), a track wide enough for full-size cylinder – rather than down-sized cylinders - can be established in one pass. Objective To demonstrate coring the scarred corpora cavernosa using Shaeer’s Cavernotome in cases of post-priapism cavernous fibrosis, enabling the implantation of a regular size inflatable implant. Methods Surgery was performed in 11 cases of post-priapism fibrosis, average duration post-priapism being 9.7 months +/−3. Incision was penoscrotal. Dilatation with blunt dilators failed. Corpora cavernosa and crura were fully excavated using Shaeer’s cavernotome, followed by dilatation of the track up to the maximum possible Hegar size with blunt dilators. Results Excavation succeeded up to size 13.5 Hegar in 7 cases and up to size 12.5 in 4 cases. One proximal crossing over occurred. No perforations were encountered. Three-piece inflatable penile prosthesis was implanted in all cases, with regular cylinders rather than down-sized. Average operative time was 65 minutes +/−12.2. Conclusions Shaeer’s Cavernotome renders penile prosthesis implantation in fibrotic cases safer and easier, and enables implantation of regular sized inflatable cylinders rather than down-sized ones. Disclosure No.

Abstract Background Revascularization with endovascular therapy (EVT) for complex below-the-knee (BTK) chronic total occlusion (CTO) remains a challenging problem. The Japanese-BTK (J-BTK) CTO score is reported as an indicator of the difficulty of BTK CTO, with the guidewire (GW) passage success rate decreasing as the grade increases. We previously reported an effective GW crossing method for the intravascular ultrasound (IVUS)-guided parallel wiring of complex BTK CTO. In this study, we investigated the feasibility of EVT using IVUS-guided wiring for BTK CTO. Materials and methods This single center, retrospective study analyzed 65 consecutive BTK CTO vessels in which IVUS-guided wiring was attempted after the failure of a conventional antegrade wiring approach from November 2020 to November 2022. The primary endpoint was the clinical success of the target CTO vessel. The secondary endpoints were the GW success rate per grade based on the J-BTK CTO score, number of GW used for CTO crossing, fluoroscopy time, and complications. Results Target vessels were the anterior tibial artery (66.2% of cases), peroneal artery (9.2%), and posterior tibial artery (24.6%). Blunt type CTO entry was performed in 55.4% of cases, calcification of entry was observed in 24.6% of cases, the mean occlusion length was 228.2 ± 93.7 mm, mean reference vessel diameter was 2.1 ± 0.71 mm, and outflow was absent in 38.5% of cases. J-BTK CTO scores of 0/1 (grade A), 2/3 (grade B), 4/5 (grade C), and 6 (grade D) were seen in 18.5%, 43.1%, 36.3%, and 1.5% of cases, respectively. The clinical success rate was 95.4%. The GW success rate by J-BTK CTO grade was as follows: grade A (100%), B (100%), C (91.7%), and D (0%). The mean number of GW used was 3.4 ± 1.4, the mean fluoroscopy time was 72.3 ± 32.5 min, and complications occurred in 7.7% of cases. Conclusion This study showed a very high clinical success rate despite the difficulty of BTK CTO. IVUS-guided EVT might be a feasible strategy for complex BTK CTO.

Introduction: Previous work from our group demonstrated that leptin is a major contributor to obesity-associated cardiovascular disease, in females, by acting on the adrenal gland to promote aldosterone production. Hypothesis: This leptin-mediated aldosterone secretion is dependent on the presence of the long form leptin receptor (LepRb) in adrenal aldosterone synthase (AS) expressing cells. Methods: We generated AS LepRb deficient mice, ASKO and WT control (ASWT), by crossing LepRb flox/flox mice with AS-Cre mice to investigate the role of LepRb in AS expressing cells in leptin-mediated cardiovascular alteration in females and also generated db/AS + mice that have LepRb only in AS expressing cells by crossing LepR TBflox/flox (db/db) with AS-Cre mice to compare their phenotype. Results: Under baseline conditions, ASKO female mice exhibited no alteration in adrenal AS, plasma aldosterone, systolic blood pressure, and aortic endothelium-dependent relaxation. To mimic obese conditions, we submitted ASWT and ASKO to chronic leptin infusion for a week. Leptin increased adrenal AS expression, aldosterone levels, and blood pressure and impaired endothelial function, which were further elevated in ASKO compared to ASWT, suggesting protective effects of AS LeRb. Both mineralocorticoid receptor (MR) antagonist. eplerenone, or AS antagonist, fadrazole, rescued endothelial function in ASWT and ASKO mice treated with leptin, indicating that leptin-mediated endothelial dysfunction is still aldosterone-dependent in ASKO mice. Restoration of LepRb in AS expressing cells in db/db decreased adrenal AS and plasma aldosterone and improved endothelial function further indicating that AS LepRb blunts leptin-mediated aldosterone production. On a molecular level, leptin treatment elevated aorta NOX1 expression in ASWT, which was further increased in ASKO. Aldosterone treatment increased NOX1 expression in cultured microvascular endothelial cells. Incubation of aortic rings with a selective NOX1 inhibitor restored endothelial function in both ASWT and ASKO treated with leptin, suggesting leptin-mediated aldosterone increased NOX1 expression in the aorta, which causes endothelial dysfunction. Conclusion: These data indicate that AS LepRb protects from leptin-mediated aldosterone production and vascular dysfunction by mitigating the effects of leptin on aldosterone production and suggests that other isoforms of LepR are involved in stimulating aldosterone production.

Introduction: NADPH oxidase (NOX) 1 but not NOX4-dependent oxidative stress plays a role in diabetic vascular disease, including atherosclerosis. Endothelin (ET)-1 has been implicated in diabetes-induced vascular complications. We showed that crossing mice overexpressing ET-1 selectively in endothelium (eET-1) with apolipoprotein E knockout ( Apoe -/- ) mice exaggerated high-fat diet-induced atherosclerosis in part by increasing oxidative stress. We hypothesized that ET-1 overexpression in the endothelium would exaggerate diabetes-accelerated atherosclerosis through a mechanism involving NOX1 but not NOX4. Methods: Six-week-old male Apoe -/- mice, eET-1/ Apoe -/- and eET-1/ Apoe -/- mice deficient in Nox1 (eET-1/ Apoe -/- / Nox1 y/- ) or Nox4 (eET-1/ Apoe -/- / Nox4 -/- ) were rendered diabetic with 55 mg/kg/day streptozotocin (STZ) IP for 5 days and studied 14 weeks later. Aortic atherosclerotic lesions were quantified using Oil Red O staining. Monocyte/macrophage infiltration and alpha-smooth muscle actin area were determined by immunofluorescence in aortic atherosclerotic lesions. Plasma cholesterol, HDL and triglycerides were measured. Results: ET-1 overexpression exaggerated 2.5-fold the atherosclerotic lesion area of the aortic sinus in diabetic Apoe -/- mice (plaque area [x10 5 μm 2 ]: 5.3±0.2 vs 2.1±0.4, P <0.05), which was reduced ~35% by Nox1 (3.5±0.4 x10 5 μm 2 , P <0.05) but not Nox4 knockout (5.0±0.7 x10 5 μm 2 ). Monocyte/macrophage infiltration was reduced ~30% in diabetic eET-1/ Apoe -/- and eET-1/ Apoe -/- / Nox4 -/- mice (31±1 and 35±2 vs 48±5% of lesion area, P <0.05) but not eET-1/ Apoe -/- / Nox1 y/- mice (35±2%). ET-1 overexpression decreased alpha-smooth muscle actin content by ~35% (9±1 vs 14±2% of lesion area, P <0.05), which was blunted by Nox1 (15±2%, P <0.05) but not Nox4 knockout (9±1%). Plasma triglycerides were unaffected by ET-1 overexpression (3.4±0.3 vs 3.6±0.5 mmol/L) but reduced by Nox1 and Nox4 knockout (2.2±0.4 and 1.8±0.4 mmol/L, P <0.05). Plasma HDL and cholesterol were similar between groups. Conclusions: Endothelium ET-1 overexpression exaggerates diabetes-accelerated atherosclerosis and reduces plaque stability through NOX1.

The pediatric population suffers from increased incidence of both major types of diabetes in recent decades, highlighting the urgent need for discovering mechanisms that regulate functional β-cell mass. We found that the master antioxidant regulator, Nrf2, is required for adaptive adult β-cell expansion under hypercaloric conditions via regulation of β-cell proliferation, β-cell survival, and maintenance of β-cell identity. Additionally, we found that Nrf2 expression in β-cells of 7-day-old mice is 15-fold higher of that in adult β-cells (p<0.0001, n=6). Based on these observations, we hypothesized that Nrf2 regulates β-cell proliferation and survival at early stages of life and contributes to the maintenance of normal β-cell mass later in life. To test this, we deleted Nrf2 in β-cells by crossing InsCre with Nrf2lox/lox mice (βNrf2KO). βNrf2KO mice at 28 days of age exhibited a 65% reduction of β-cell mass compared to control mice (p<0.0005, n=6). Importantly, at 7 days of age, βNrf2KO mice displayed a 65% reduction of β-cell proliferation (p<0.0001, n=6), a 22% reduction of nuclear Pdx1 levels (p<0.0001, n=6), a 5.84-fold increase in β-cell death (±0.04, p<0.001, n=6) and a 2.93-fold increase in β-cell oxidative stress (p<0.001, n=6) compared to control mice. Surprisingly, daily administration of antioxidant NAC to 7-day-old βNrf2KO mice only partially restored β-cell proliferation but completely blunted oxidative stress compared to control mice. This suggests that neonatal β-cell proliferation induced by Nrf2 relies on additional mechanisms beyond oxidative stress. Interestingly, RNAseq of isolated islets from 7-day-old βNrf2KO mice showed decreased expression of mitochondrial encoded genes and upregulation of stress-responsive genes. This suggests that Nrf2 controls β-cell proliferation by regulating mitochondrial bioenergetics. We conclude that Nrf2 is required for β-cell mass expansion in neonatal ages by controlling β-cell proliferation, identity, and survival. Disclosure S.Baumel-alterzon: None. L.S.Katz: None. L.Lambertini: None. A.Garcia-ocana: Consultant; Sun Pharmaceutical Industries Ltd. D.Scott: None. Funding National Institutes of Health (DK128387, DK114338)

Our previous studies have shown that renal medullary sphingosine-1-phosphate (S1P) stimulates urinary sodium excretion via the S1P type 1 receptor (S1PR1), which primarily located in the collecting ducts, and that knockout of S1PR1 in the collecting duct leads to sodium retention and enhanced salt-sensitive hypertension. Preliminary studies using immunostaining showed that acid ceramidase (AC), the upstream rate-limit enzyme for S1P production, was primarily present in collecting ducts in the renal medulla and predominantly in principal cells. The present study tested the hypothesis that the deletion of AC in collecting duct impairs the renal sodium excretion and promotes salt-sensitive hypertension. Mice with collecting duct (CD)-specific knockout of AC (CD-AC-KO) were generated by crossing the AC-encoding gene Asah1-floxed mice with aquaporin 2-Cre mice. Male adult mice were used. High salt intake significantly increased protein levels of AC by 2-fold as well as the S1P levels by 1-fold in the renal medulla in control mice, whereas the high salt-induced increase in S1P levels were substantially inhibited by > 50% in CD-AC-KO mice. Functionally, pressure natriuresis was blunted by 65% and the positive salt balance induced by chronic high salt intake was doubled in CD-AC-KO mice compared with control mice. These results indicated the impairment of renal medullary function and the promotion of sodium retention in CD-AC-KO mice. The treatment of a low dose deoxycorticosterone acetate (DOCA) and high salt intake for 2 weeks produced more significant increase in MAP in CD-AC-KO mice compared with control Mice (136±7mmHg vs 109±4 mmHg). These findings demonstrated that deletion of AC in collecting duct reduced S1P levels, inhibited sodium excretion, promoted sodium retention and accelerated DOCA-salt-induced salt-sensitive hypertension, suggesting that the AC in collecting duct is an important anti-hypertensive pathway by promoting sodium excretion and that impairment of renal medullary AC may represent a novel mechanism for salt-sensitive hypertension. R01DK140219, R01HL145163 This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.

Primary cilia possess an array of receptors and signaling from this organelle is critical for neuronal function. BBS1 is an important component of a receptor trafficking complex (BBSome) that facilitates signaling in both primary cilia and plasma membrane. Humans harboring BBS1 mutations develop obesity and type 2 diabetes. Recent studies from our lab and others have shown that BBS1 deletion in hypothalamic neurons lead to obesity and glucose dysregulation. However, the role of hindbrain BBS1 in metabolic control is unknown. We hypothesized that brainstem BBS1 regulates metabolism through primary cilia function. As an initial step towards testing this hypothesis, we deleted BBS1 selectively from hindbrain neurons by crossing Phox2bCre mice with BBS1fl/fl mice and assessed metabolic parameters. Cre expression in hindbrain nuclei was validated by crossing these mice on a Cre-dependent tdTomato background. Both male and female Phox2bCre/BBS1fl/fl mice fed a chow diet exhibited reduced body weight (P<0.05) during adolescence (5-8 weeks old) but had normal weights in adulthood (9-20 weeks) relative to control littermates. In both sexes, hindbrain BBS1 deletion impaired glucose clearance at 20 weeks of age, independent of insulin sensitivity, as indicated by elevated area under the curve (AUC) in glucose (male AUC 31368±1986 vs. 25497±1834, female AUC 22643±1427 vs. 19304±800, P<0.05), but not insulin (male AUC 11505±905 vs. 10416±680, female AUC 8314±333 vs. 9098±525), tolerance test. Interestingly, male Phox2bCre/BBS1fl/fl mice maintained on an obesogenic (high fat-sucrose) diet displayed blunted weight gain during the first 14 weeks of age (P<0.05) which tended to be reduced relative to controls up to 20 weeks of age. Remarkably, NMR-based body composition revealed reduced lean (22.9±0.6 vs. 24.9±0.6 g, P<0.05), but not fat (23.8±1.4 vs. 23.7±1.2 g), mass in diet-induced obese adult Phox2bCre/BBS1fl/fl mice. These data suggest that hindbrain BBS1 is important for weight gain during adolescence and glucose handling in both male and female mice. Moreover, these results suggest that brainstem BBS1 negatively regulates weight gain induced by a high fat/sucrose diet. Since BBS1 has functions independent of cilia, we directly tested the role of hindbrain primary cilia on metabolic homeostasis. For this, we assessed how brainstem ablation of cilia through deletion of IFT88 protein (Phox2bCre/IFT88fl/fl mice) affect metabolic parameters. Male, but not female, Phox2bCre/IFT88fl/fl animals displayed reduced body weight during adolescence (P<0.05). Glucose tolerance test showed normal glucose regulation in adulthood (male AUC 28565±2007 vs. 28762±2321, female AUC 23820±836 vs.22601±175). Collectively, these data show a sex-dependent role for brainstem primary cilia in body weight gain during adolescence. Furthermore, our findings reveal that the critical role of hindbrain BBS1 in glucose homeostasis is independent of its function in primary cilia. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Cdc42 is a Ras-related GTPase that plays an important role in regulation of actin cytoskeletal architecture. Blocking the ability of Cdc42 to activate its effectors has been shown to inhibit a range of cellular functions including cell polarization, migration, proliferation and differentiation. Consistent with its critical roles in vitro, the inactivation of Cdc42 in mice resulted in embryonic lethality before E6.5. The early embryonic lethal phenotype of Cdc42-null mice has made it not useful for studies on the interesting questions of the roles and mechanisms of Cdc42 in the vascular development. To overcome this problem, we have generated an endothelial cell (EC) specific Cdc42 knockout mouse line by crossing Cdc42/flox mice with vascular endothelial cadherin Cre mice. Our results have demonstrated that the deletion of Cdc42 in ECs resulted in embryonic lethality with severe edema. Whole mount immunofluorescence staining showed mesentery collecting lymphatic vessel maturation and valve formation defects. Interestingly, lymphangiogenesis in the intestine wall was totally disrupted in Cdc42 EC knockout embryos. Moreover, we analyzed the role of Cdc42 in lymphatic vessel formation in the skin. We found that Cdc42 is required for tip cells filopodia formation and the deletion of Cdc42 in endothelial cells impaired lymphatic vessel sprouting, branching and the mutant lymphatic vessels displayed blunt-ended, bulbous lymphatic. We also noted that the size of lymphatic lumen was significantly increased. Taken together, our data suggested that Cdc42 plays an essential role in lymphatic branching, maturation and valve formation during development.

Purpose: To assess the surgical outcomes of transconjunctival approach in management of different orbital tumors at different locations. Methods: This prospective, non-comparative, clinical interventional study was conducted in the period between March 2017 and January 2020 and included 61 patients with histologically proved orbital tumors. In all cases, a conjunctival incision near the fornix was made depending on the tumor location as revealed by CT or MRI. A traction suture was applied to one or two relevant recti muscles to guide the globe toward the desired direction. Blunt orbital dissection was made toward the tumor until exposing its anterior surface. The procedure was considered successful if the predetermined decision (total excision with improved clinical manifestations for benign and biopsy for diagnosis in malignant tumors) was achieved without causing permanent complications. The procedure was considered a failure if the predetermined decision was not achieved or if permanent complications developed. Results: The patients were divided into: Group A of 47 patients (77.05%) with benign tumors and Group B of 14 patients (22.95%) with malignant tumors. The overall success rate of the approach was 98.36% (60 out of 61 patients), while failure occurred in one case (1.64%). Conclusions: The transconjunctival orbitotomy is an excellent approach to manage different tumors at different orbital locations with rapid recovery and maximum cosmetic results. It is the only approach that can access intra-conal, mid-orbital tumors whatever their relation to the optic nerve without crossing it.

Abstract Purpose To present a safety-optimized ultrasound-guided minimal invasive carpal tunnel release (CTR) procedure. Materials and Methods 104 patients (67 female, 37 male; mean age 60.6 ± 14.3 years, 95% CI 57.9 to 63.4 years) with clinical and electrophysiological verified typical carpal tunnel syndrome were referred for a high-resolution ultrasound of the median nerve and were then consecutively assigned for an ultrasound-guided CTR after exclusion of possible secondary causes of carpal tunnel syndrome such as tumors, tendovaginitis, ganglia and possible contraindications (e.g., crossing collateral vessels, nerve variations). Applying a newly adapted and optimized algorithm, basing on the work proposed by Petrover et al. CTR was performed using a button tip cannula which has several safety advantages: On the one hand, the button tip cannula acts as a blunt and atraumatic guiding splint for the subsequent insertion of the hook-knife, and on the other hands, it serves as a “hydro-inflation”-tool, i.e., a fluid-based expansion of the working-space is warranted during the whole procedure whenever needed. Results In all patients, successful releases were confirmed by the depiction of a completely transected transverse carpal ligament during and in the postoperative ultrasound-controls two weeks after intervention. All patients reported markedly reduction of symptoms promptly after this safety-optimized ultrasound-guided minimal invasive CTR and at the follow-up examination. No complications were evident. Conclusion The here proposed optimized algorithm assures a reliable and safe ultrasound-guided CTR and thus should be taken into account for this minimal invasive interventional procedure.

Abstract Introduction Metabolic syndrome (MetS) is well known as the risk of cardiovascular diseases associated with endothelial dysfunction and induces steatohepatosis. Insulin resistance is a major character of MetS, which affects intracellular signaling pathways and endothelial function. Extracellular signal-regulated kinase (ERK) is a major component of insulin signal and many of vasoactive peptides, which were released in MetS, can activate it in endothelium. However, the role of endothelial ERK in nitric oxide (NO) bioactivity in MetS in in vivo has been unknown. Purpose The aim of this study is to clarify the role of endothelial ERK2 on NO bioactivity in mice model of MetS. Methods and results We created endothelial specific ERK2 knock out mice (EE2KO) crossing Tie2-Cre mice and ERK2 flox mice and fed them with normal or high-fat/high-sucrose diet (HFHSD) for 24 weeks. Serum glucose and insulin levels and HOMA-IR were lowered in EE2KO with HFHSD without changing body weight. In wild type mice (WT) with HFHSD, nonalcoholic fatty liver disease (NAFLD) activity score, fibrosis score and serum ALT level were increased, all of which were blunted in EE2KO. EE2KO with HFHSD lowered systolic blood pressure (WT: 123.7±5.83 mmHg, EE2KO: 101.4±3.66 mmHg, P<0.01, N=8) without changing heart rate, which was increased to the same levels with L-NAME, an endothelial NO synthase inhibitor, in both groups. Serum NO levels measured with serum nitrite/nitrate concentrations were increased in EE2KO with HFHSD (WT: 23.10±3.74 μmol/l, EE2KO: 41.71±6.73 μmol/l, P<0.05, N=12). Endothelial function was assessed with the isometric tension measurement of aortic rings with acetylcholine (ACh). ACh-induced relaxation was improved in EE2KO with HFHSD. Superoxide production of aorta from EE2KO was lowered than WT with HFHSD in dihydroethidium (DHE) staining. S18886, an antagonist of the thromboxane A2-prostanoid (TP) receptor, decreased superoxide production of aorta in DHE staining resulting in improving endothelial function in the isometric tension measurement of aortic rings. Oral administrations of S18886 decreased systolic blood pressure, serum fasting glucose and insulin levels, and surprisingly improved steatohepatosis by decreasing NAFLD activity score and fibrosis score. Relaxation of aortic rings with ACh Conclusions Endothelial ERK2/TP receptor pathway increases superoxide production and decreased NO bioactivity, resulting in deteriorating endothelial function, insulin resistance and steatohepatosis, which were improved by antagonist of the TP receptor in mice model of MetS. The present study indicates that ERK2/TP pathway could be a therapeutic target for complications of MetS.

Abstract Introduction Endothelial dysfunction (ED) is a key underpinning of cardiovascular disease in obesity, but the underlying molecular mechanisms remain elusive. Neurofibromin 2 (NF2) is a scaffold-like protein implicated in various cellular processes, namely growth, differentiation and survival. NF2 is inactivated by Akt-dependent phosphorylation at Ser518, whereas its dephosphorylation by the myosin phosphatase target subunit 1 (MYPT-1) leads to an active conformation. The role of NF2 in obesity-related alterations of endothelial phenotype remains elusive. Purpose To investigate whether NF2 participates to ED in obesity. Methods Human aortic endothelial cells (HAECs) were exposed to palmitic acid (PA, 200 uM) or vehicle for 48 hours. Gene silencing of NF2 was performed by small interfering RNA (siRNA). Protein expression was assessed by Western blot. Nitric oxide (NO) levels were measured by using a colorimetric assay. The interaction of NF2 with endothelial proteins was investigated by co-immunoprecipitation. To specifically determine NF2 role in the endothelium, we generated mice with endothelium-specific deletion of NF2 (NF2 ECKO) by crossing NF2flox/flox mice with tamoxifen-inducible endothelial-specific cre mice [Cdh5(PAC)-CreERT2]. Endothelium-dependent relaxations to acetylcholine (Ach, 10–9 to 10–5 mol/L) were assessed in aortas isolated from male NF2 ECKO and wild type littermates, fed a control (10 kcal% fat) and a high fat diet (60 kcal% fat) for 20 weeks. NF2 signalling and endothelial function were also assessed in small visceral fat arteries (VFA) isolated from 18 obese and 18 age-matched healthy subjects undergoing bariatric surgery and cholecystectomy, respectively. Results Exposure of HAECs to PA decreased NF2 phosphorylation at Ser518, thus leading to an active protein conformation. Blunted NF2 phosphorylation was explained by a reduction of Akt phosphorylation at Ser473 and a concomitant increase of MYPT-1 phosphorylation at Thr696. Pull-down experiments revealed that NF2 binds and activates Caveolin 1 (Cav-1), a pivotal repressor of endothelial NO synthase (eNOS). NF2 knockdown in PA-treated HAECs prevented eNOS–Cav-1 interaction, thus preserving eNOS activity and NO levels. In aortas from obese mice, we found that NF2-Cav-1 interaction was responsible for impaired eNOS activity, reduced NO levels and endothelial dysfunction. By contrast, Ach-dependent vasorelaxation were preserved in obese mice with endothelium-specific deletion of NF2. Moreover, we found that NF2 is activated in VFA from obese patients as compared to healthy controls, and its activity negatively correlated with Ach-dependent vasorelaxation of isolated VFA, as assessed by organ chamber experiments. Conclusions The present findings – obtained in human endothelial cells, conditional mouse models and visceral fat arteries from obese patients – suggest that targeting NF2 may represent a potential therapeutic strategy to prevent ED in patients with obesity.

Background: Endothelial progenitor cells (EPC) are both reduced and dysfunctional in hypertension that correlates inversely with its mortality, but the mechanisms are poorly understood. eNOS critically regulates EPC mobilization and function, but is uncoupled in DOCA-salt hypertension due to reduced essential cofactor tetrahydrobiopterin (BH4). We tested the hypothesis that GTP cyclohydrolase I (GTPCH), the rate-limiting enzyme of BH4 synthesis, protects EPC function in DOCA-salt mice. Methods and Results: EPCs were isolated from peripheral blood and bone marrow of wild-type (WT), WT DOCA-salt, endothelial-specific GTPCH transgenic (Tg-GCH), DOCA-salt treated Tg-GCH, and BH4 deficient hph-1 mice. Both EPC adhesion and angiogenic functions were impaired in WT DOCA (58 and 38%, respectively, n=5– 8, p<0.01) and hph-1 mice (40 and 31%, respectively, n=5–8, p<0.01, vs. WT), which were rescued in DOCA Tg-GCH mice (93 and 92%, respectively, n=6–7). Superoxide (O 2 − , DHE-) and NO (DAF-FM flow cytometry) levels in EPCs were elevated and reduced, respectively, in WT DOCA (220% and 53%, respectively, n=5–12, p<0.01) and hph-1 mice (280% and 41%, respectively, n=6–12, p<0.05, vs. WT), which were blunted in DOCA Tg-GCH mice (137% and 96%, respectively, n=6–7). Thrombospondin-1 (TSP-1), a secreted potent anti-angiogenic protein, was elevated in EPCs of WT DOCA and hph-1 mice (290 and 280%, respectively, n=8–12, p<0.01 vs. WT), but not in DOCA Tg-GCH mice (160%, n=9–12, p>0.05). In vitro treatment of EPCs from WT DOCA mice with BH4, PEG-SOD, or L-NNA significantly reduced TSP-1 (51, 42 and 53%, respectively, n=5–12, p<0.01) and O 2 − levels (58, 67 and 69%, respectively, n=5– 6, p<0.01 vs. WT-DOCA). NO level in EPCs of WT DOCA mice was rescued by BH4 and PEG-SOD (144 and 165%, respectively, n=4–7, p<0.05 vs. control), but not L-NNA. Crossing eNOS KO and Tg-GCH mice resulted in preserved circulating EPC number (88%, n=4–15, p>0.05 vs. WT), reduced EPC O 2 − level (64%, n=4 – 8, p<0.05 vs. WT-DOCA), and improved EPC adhesion and angiogenic functions (160 and 158%, respectively, n=4 –12, p<0.05 vs. WT-DOCA). Conclusion: We demonstrate for the first time that GTPCH/BH4 pathway protects EPC function via suppressing oxidative stress and TSP-1 level in salt-sensitive hypertension. This research has received full or partial funding support from the American Heart Association, Midwest Affiliate (Illinois, Indiana, Iowa, Kansas, Michigan, Minnesota, Missouri, Nebraska, North Dakota, South Dakota & Wisconsin).

Vascular support structures are an important tool for treating valve stenosis. A large population of patients are treated for valvular disease and the principal mode of treatment is the use of percutaneous valvuloplasty. Stent devices are proving to be an improved treatment method; these devices now account for 20% of treatments in Europe. This new technology provides highly effective results at minimal cost and short duration of hospitalization. Accurate and reliable structural analysis provides essential information to the design team in an environment where in vivio experimentation is extremely expensive, or impossible. This paper describe the design of vascular support structure (stents), to provide designers with estimates of the critical parameters which are essential to restore the functions of the endothelium of the Aorta during and after implantation without injuring it. Stent geometries were uniquely defined using the following parameters. (a) Diameter of the aorta; (b) Distance between the aortic root and the coronary artery roots; (c) Position of the coronary arteries; (d) Diameter of the coronaries; (e) Stent–Endothelium Mechanics. Keeping these parameters into consideration a novel stent model was designed to suit its requirement for percutaneous replacement. The 3D geometry of the repeatable units of the stent was generated using SOLIDWORKS modeling software. Using the repeating unit geometry of each stent design, solid models were generated. The unit consisted of 8 lips with two non crossing struts making a circular diameter of 16 mm at the center and 18 mm at either ends. The upper and the lower portions of the prosthesis has a high radial force, the upper portion flared to fix the stent firmly in the ascending aorta and the lower portion to expand against the calcified leaflets and to avoid recoil. The middle portion which bears the valve is constrained and narrower to avoid obstruction of the coronary arteries. This varying diameter of different parts of the stent creates the blunt hooks at either end of the stent. The methodology described in this paper is proposed as a method to compare and analyze the existing stents and the ones proposed here. However, further analysis and studies are needed before these stents are fabricated and deployed. Animal experiments are being planned currently for this purpose.

Nitric oxide (NO) released from NO synthase 1 (NOS1) which is highly expressed in the macula densa (MD), blunts tubuloglomerular feedback (TGF) response and dilates the afferent arteriole. However, the role of MD NOS1 and TGF in regulation of chronic salt-water balance and blood pressure is unknown. We developed a strain of tissue-specific NOS1 knockout mice to examine the significance of NOS1 in the MD. We first developed a strain of MD Cre mice using the sodium, potassium, 2 chloride cotransporter (NKCC2) promoter. The NKCC2 promoter was cloned into the plasmid pBS594 to create the pNKCC2-Cre transgene. To characterize this new Cre strain, we crossed the NKCC2-Cre mice with an EGFP reporter strain (Rosa). Cre activity measured by EGFP fluorescence was found in MD and was further confirmed via immunofluorescent staining in isolated perfused MD and found to be limited to the MD and not in the distal tubule, indicating a valid NKCC2-Cre strain. Next, we generated tissue specific NOS1 knockout mice by crossing NKCC2-Cre mice with NOS1 flox/flox mice, which target the exon-6 of NOS1, therefore deletes all splice variants of NOS1. We confirmed NOS1 KO by immunohistochemistry and found no staining of NOS1 in the MD in KO mice, while NOS1 was abundant in the MD of WT mice. To measure NO generation in the MD, we perfused isolated MD and loaded with fluorescent dye DAF-2. When we switch tubular NaCl from 10 to 80 mM, NO generation in the MD was from 114 ± 12 to 165 ± 11 units/min in WT mice, while in KO mice NO were absent at both 10 and 80 mM NaCl. (n=5; p<0.01) To measure TGF response, we measured maximal change of stop flow pressure using micropuncture. In WT mice, TGF was 4.5 ± 0.3 mmHg; while in KO mice, TGF was 8.4 ± 0.7 mmHg. (p<0.05) To determine the effect of MD NOS1 on the blood pressure response to salt intake , we measured mean arterial pressure (MAP) by telemetry in mice fed a low salt diet (0.01% NaCl) for 10 days, followed by a high salt diet (8%NaCl) for 14 days. MAP of KO mice increased by 10.5 ± 2.4; while WT mice increased 3.1± 2.7 mmHg (p <0.01, n=5). These results indicate that NOS1 in the MD and TGF play important roles in the regulation of blood pressure in response to changes in sodium intake.


 
 
 i) In Australia we do a lot of thinking about home. Or so it would seem from all the talk about belonging, home, being at home (see Read). A sure sign of displacement, some might say. In his recent memoir, John Hughes writes: It is a particularly Australian experience that our personal heritage and sense of identity includes a place and a history not really our own, not really accessible to us. The fact that our sense of self-discovery and self-realisation takes place in foreign lands is one of the rich and complex ironies of being Australian. (24-25) My sense of self-discovery did not occur in a foreign land. However, my personal heritage and sense of identity includes places and histories that are not really my own. Unlike Hughes I don’t have what is often portrayed as an exotic heritage; I am plainly white Australian. I grew up on the Far North Coast of New South Wales, on farms that every year knew drought and flood. My place in this country – both local and national – seemingly was beyond question. I am after all a white, settler Australian. But I left Kyogle twenty years ago and since then much has changed. My project is very different than Hughes’. However, reading his memoir led me to reflect upon my sense of belonging. What is my home made from? Like Hughes I want to deploy memories from my childhood and youth to unpack my idea of home. White settler Australians’ sense of belonging is often expressed as a profound feeling of attachment; imagined as unmediated (Moreton-Robinson 31). It is a connection somehow untroubled by the worldliness of the world: it is an oasis of plentitude. For Indigenous Australians, Aileen Moreton-Robinson argues, non-Indigenous Australians sense of belonging is tied to migrancy, while the Indigenous subject has an ontological relationship to land and these modes are incommensurable (31). Since colonisation the nation state has attempted through an array of social, legal, economic and cultural practices to break Indigenous people’s ontological connections to land, and to cast them as homeless in the ‘modern’ world. The expression of belonging as a profound sense of attachment – beyond the material – denies not only the racialised power relations of belonging and dispossession, but also the history of this sentiment. This is why I want to stay right here and take up Moreton-Robinson’s challenge to further theorise (and reflect) upon how non-Indigenous subjects are positioned in relation to the original owners not through migrancy but through possession (37). ii) Australia has changed a lot. Now most understand Australia to be comprised of a plurality of contradictory memories, imaginaries and histories, generated from different cultural identities and social bodies. Indigenous Australians, who have been previously spoken for, written about, categorised and critiqued by non-Indigenous people, have in the last three decades begun to be heard by mainstream Australia. In a diversity of mediums and avenues Indigenous stories, in all their multiplicity, penetrated the field of Australian culture and society. In so doing, they enter into a dialogue about Australia’s past, present and future. The students I teach at university arrive from school with an awareness that Australia was colonised, not discovered as I was taught. Recent critical historiography, by both Indigenous and non-Indigenous writers and academics, calls for and creates a new Australian memory (Hage 80). A memory, or memories, which the reconciliation movement not only want acknowledged by mainstream Australia but also integrated into national consciousness. Over the last twenty years, many Australian historians have reinforced the truths of fictional and autobiographical accounts of colonial violence against Indigenous people. The benign and peaceful settlement of Australia, which was portrayed in school history lessons and public discourse, began to be replaced by empirical historical evidence of the brutal subjugation of Indigenous people and the violent appropriation of Indigenous land. Indigenous struggles for recognition and sovereignty and revisionist history have created a cultural transformation. However, for all the big changes there has been limited investigation into white Australians’ sense of belonging continuing to be informed and shaped by settler colonial desire. Indigenous memories not only contest and contradict other memories, but they are also derived from different cultural bodies and social and historical contexts. My memory of our farm carved out of Toonumbah State Forest is of a peaceful place, without history; a memory which is sure to contradict Bundjalung memories. To me Kyogle was a town with only a few racial problems; except for the silences and all those questions left unasked. Ghassan Hage argues that a national memory or non-contradictory plurality of memories of colonisation in Australia is impossible because although there has been a cultural war, the two opposing sides have not assimilated to become one (92). There remain within Australia, ‘two communal subjects with two wills over one land; two sovereignties of unequal strength’ (Hage 93). The will of one is not the will of the other. I would argue that there is barely recognition of Indigenous sovereignty by non-Indigenous Australians; for so many there is only one will, one way. Furthermore, Hage maintains that: For a long time to come, Australia is destined to become an unfinished Western colonial project as well as a land in a permanent state of decolonisation. A nation inhabited by both the will of the coloniser and the will of the colonised, each with their identity based on their specific understanding, and memory, of the colonial encounter: what was before it and what is after it. Any national project of reconciliation that fails to fully accept the existence of a distinct Indigenous will, a distinct Indigenous conatus, whose striving is bound to make the settlers experience ‘sadness’, is destined to be a momentary cover-up of the reality of the forces that made Australia what it is. (94) Why must Indigenous will make settlers experience sad passions? Perhaps this is a naïve question. I am not dismissing Hage’s concerns, and agree with his critique of the failure of the project of reconciliation. However, if we are to understand the forces that made Australia what it is – to know our place – then as Hage writes we need not only to acknowledge these opposing forces, but understand how they made us who we are. The narrative of benign settlement might have resulted in a cultural amnesia, but I’m not convinced that settler Australians didn’t know about colonial violence and its aftermath. Unlike Henry Reynolds who asked ‘why didn’t we know?’ I think the question should be, as Fiona Nicoll asks, ‘what is it we know but refuse to tell?’ (7). Or how did I get here? In asking what makes home, one needs to question what is excluded to enable one to stay in place. iii) When I think of my childhood home there is one particular farm that comes to mind. From my birth to when I left home at eighteen I lived in about six different homes; all but one where on farms. The longest was for about eight years, on a farm only a few kilometres from town; conveniently close for a teenager wanting all the ‘action’ of town life. It was just up the road from my grandparents’ place, whose fridge I would raid most afternoons while my grandmother lovingly listened to my triumphs and woes (at least those I thought appropriate for her ears). Our house was set back just a little from the road. On this farm, my brother and I floated paper boats down flooded gullies; there, my sisters, brother and I formed a secret society on the banks of the picturesque creek, which was too quickly torn apart by factional infighting. In this home, my older sisters received nightly phone calls from boys, and I cried to my mother, ‘When will it be my turn’. She comforted me with, ‘Don’t worry, they will soon’. And sure enough they did. There I hung out with my first boyfriend, who would ride out on his motor bike, then later his car. We lolled around on our oddly sloping front lawn and talked for hours about nothing. But this isn’t the place which readily comes to mind when I think of a childhood home. Afterlee Rd, as we called it, never felt like home. Behind the house, over the other side of the creek, were hills. Before my teens I regularly walked to the top of the first hill and rode around the farm, but not all the way to the boundary fence. I didn’t belong there. It was too exposed to passing traffic, yet people rarely stopped to add to our day. For me excitement and life existed elsewhere: the Gold Coast or Lismore. When I think of my childhood home an image comes to mind: a girl child standing on the flat between our house and yards, with hills and eucalypts at her back, and a rock-faced mountain rising up behind the yards at her front. (Sometimes there is a dog by her side, but I think it’s a late edition.) The district was known as Toonumbah because of its proximity (as the crow flies) to Toonumbah Dam. My siblings and I ventured across the farm and we rode with my father to muster, or sometimes through the adjoining State Forest to visit our neighbours who lived deep in the bush. I thought the trees whispered to me and watched over us. They were all seeing, all knowing, as they often are for children – a forest of gods. Sometime during my childhood I read the children’s novel Z for Zachariah: a story of a lone survivor of an apocalypse saved by remaining in a safe and abundant valley, while the rest of the community went out to explore what happened (O’Brien). This was my idea of Toonumbah. And like Zachariah’s valley it was isolated and for that reason, in spite of its plenty, a strange home. It was too disconnected from the world. Despite my sense of homeliness, I never felt sovereign. My disquiet wasn’t due to a sense that at any moment we might be cast out. Quite the opposite, we were there to stay. And not because I was a child and sovereignty is the domain of adults. I don’t think, at least as a feeling, it is. But rather because sovereignty is tied to movement or crossings. Not just being in place, but leaving and returning, freely moving through and around, and welcoming others who recognise it as ‘our’ place. Home is necessitated upon movement. And my idea of this childhood home is reliant upon a romanticised, ‘profound’ feeling of attachment; a legacy of settler colonial desire. There is no place like home. Home is far more than a place, it is, as Blunt and Dowling suggest, about feelings, desire, intimacy and belonging and relationships between places and connections with others (2). One’s sense of home has a history. To be at home one must limit the chaos of the world – create order. As we know, the environment is also ordered to enable a sense of bodily alignment and integrity. How or rather with whom does one establish connections with to create a sense of home? To create a sense of order, who does one recognise as belonging or not? Who is deemed a part of the chaos? Here Sara Ahmed’s idea of the stranger is helpful. Spaces are claimed, or ‘owned’, she argues, not so much by inhabiting what is already there, but rather movement or ‘passing through’ creates boundaries, making places by giving them a value (33). Settlers moved out and across the country, and in so doing created the colonies and later the nation by prescribing an economic value to the land. Colonialism attempts to enclose both Indigenous people and the country within its own logic. To take possession of the country the colonisers attempted to fix Indigenous people in place. A place ordered according to colonial logic; making the Indigenous subject out of place. Thus the Indigenous ‘stranger’ came into view. The stranger is not simply constituted by being recognised by the other, but rather it is the recognition of strangers which forms the local (Ahmed 21-22). The settler community was produced and bounded by their recognition of strangers; their belonging was reliant upon others not belonging. The doctrine of terra nullius cleared the country not only of people, but also of the specifics of Indigenous place, in an attempt to recreate another place inspired by the economic and strategic needs of the colonisers. Indigenous people were further exposed as strangers in the ‘new’ country by not participating in the colonial economy and systems of exchange. Indigenous people’s movement to visit family, to perform ceremony or maintain connections with country were largely dismissed by the colonial culture and little understood as maintaining and re-making sovereignty. European forms of commerce made the settlers sovereign – held them in place. And in turn, this exchange continues to bind settler Australians to ways of being that de-limit connections to place and people. It created a sense of order that still constrains ideas of home. Colonial logic dominates Australian ideas of sovereignty, thus of being at home or belonging in this country. Indeed, I would argue that it enforces a strange attachment: clinging fast as if to a too absent parent or romancing it, wooing a desired but permissive lover. We don’t know, as Fiona Nicoll questions, what Indigenous sovereignty might look like. Discussions of sovereignty are on Western terms. If Indigenous sovereignty is recognised at all, it is largely figured as impractical, impossible or dangerous (Nicoll 9). The fear and forgetting of the long history of Indigenous struggles for sovereignty, Nicoll writes, conceals the everydayness of the contestation (1). Indigenous sovereignty is both unknown and too familiar, thus it continues to be the stranger which must be expelled to enable belonging. Yet without it we cannot know the country. iv) I carry around a map of Australia. It is a simple image, a crude outline of the giant landmass; like what you find on cheap souvenir tea-towels. To be honest it’s just the continent – an islandless island – even Tasmania has dropped off my map. My map is not in my pocket but my head. It comes to mind so regularly I think of it as the shape of my idea of home. It is a place shared by many, yet singularly mine. I want to say that it is not the nation, but the country itself, but of course this isn’t true. My sense of Australia as my home is forged from an imaginary nation. However, I have problems calling Australia home – as if being at home in the nation is like being in an idealised family home. What is too often sentimentalised and fetishised as closed and secure: a place of comfort and seamless belonging (Fortier 119). Making home an infantile place where everything is there for me. But we understand that nations are beyond us and all that they are composed of we cannot know. Even putting aside the romantic notions, nations aren’t very much like home. They are, however, relational. Like bower birds, we collect sticks, stones, shells and coloured things, building connections with the outside world to create something a bit like home in the imaginary nation. I fill my rough map with ‘things’ that hold me in place. We might ask, is a home a home if we don’t go outside? My idea of home borrows from Meaghan Morris. In Ecstasy and Economics, she is attempting to create what Deleuze and Guattari call home. She writes: In their sense of the term, “home does not pre-exist”; it is the product of an effort to “organize a limited space”, and the limit involved is not a figure of containment but of provisional (or “working”) definition. This kind of home is always made of mixed components, and the interior space it creates is a filter or a sieve rather than a sealed-in consistency; it is not a place of origin, but an “aspect” of a process which it enables (“as though the circle tended on its own to open into a future, as a function of the working forces it shelters”) but does not precede – and so it is not an enclosure, but a way of going outside. (92) If home is a way of going outside then we need to know something about outside. Belonging is a desire and we make home from the desire to belong. In desiring belonging we should not forsake the worldliness of the world. What is configured as outside home are often the legal, political, economic and cultural conditions that have produced contemporary Australia. However, by refusing to engage with how colonialism and Indigenous sovereignty have made Australia one might not be able to go outside; risk imprisoning oneself in a too comfortable space. By letting in some of the elements which are strange and unhomely, one might begin to build connections which aid the reimagining of the self and the social, which in turn enables one to not only live in postcolonial Australia but participate in creating it (Probyn). A strange place: unsettled by other desires, histories, knowledge and memories, but a place more like home. I am arguing that we need to know our place. But knowing our place cannot be taken for granted. We need many hearts and minds to allow us to see what is here. The childhood home I write of is not my home, nor do I want it to be. However, the remembering or rather investigation of my idea of home is important. Where has it come from? There has been a lot of discussion about non-Indigenous Australians being unsettled by revisionist historiography and Indigenous demands for recognition and this is true, but the unsettlement has been enabling. Given that settler Australians are afforded so much sovereignty then there seems plenty of room for uncertainty. We don’t need to despair, or if we do, it could be used productively to remake our idea of home. If someone were to ask that tired question, ‘Generations of my family have lived here, where am I going to go?’ The answer is no where. You’re going no where, but here. The question isn’t of leaving, but of staying well. References Ahmed, Sara. Strange Encounters: Embodied Others in Post-coloniality. London: Routledge, 2000. Blunt, Alison, and Robyn Dowling. Home. London: Routledge, 2006. Fortier, Anne-Marie. “Making Home: Queer Migrations and Motions of Attachment.” Uprootings/Regrounding: Questions of Home and Migration. Eds S. Ahmed et. al. Oxford: Berg, 2003. 115-135. Gelder, Ken, and Jane Jacobs. Uncanny Australia: Sacredness and Identity in a Postcolonial Nation. Carlton, Vic: Melbourne UP, 1998. Hage, Ghassan. Against Paranoid Nationalism. Annandale: Pluto Press, 2003. Hughes, John. The Idea of Home: Autobiographical Essays. Sydney: Giramondo, 2004. Moreton-Robinson, Aileen. “I Still Call Australia Home: Indigenous Belonging and Place in a White Postcolonizing Society.” Uprootings/Regrounding: Questions of Home and Migration. Eds S. Ahmed et. al. Oxford: Berg, 2003. 23-40. Morris, Meaghan. Ecstasy and Economics: American Essays for John Forbes. Sydney: Empress, 1992. Nicoll, Fiona. “Defacing Terra Nullius and Facing the Public Secret of Indigenous Sovereignty in Australia.” borderlands 1.2 (2002): 1-13. O’Brien, Robert C. Z for Zachariah: A Novel. London: Heinemann Educational, 1976. Probyn, Elspeth. Outside Belongings. New York: Routledge, 1996. Read, Peter. Belonging: Australians, Place and Aboriginal Ownership. Cambridge: Cambridge UP, 2000. Reynolds, Henry. Why Weren’t We Told?: A Personal Search for the Truth about Our History. Melbourne: Penguin, 2002. 
 
 
 
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Introduction: Grey Wardens, Blights, and Staying Vigilant Bodies are porous and under threat of mutation for both Grey Wardens and the darkspawn they fight in BioWare’s fantasy role-playing videogame series Dragon Age (2009-2024). If they survive the Joining Ritual, Grey Wardens become abject figures, poised in the liminal space between ‘us’ and ‘not us’. Since the Joining forges a connection to darkspawn that will also, inevitably, eventually, mutate them into a blighted creature themselves, Grey Wardens exist in a tremendously Gothic state of becoming. On the other side of this binary are the darkspawn, transgressive entities first introduced as scurrying and bipedal. Eventually, however, the player can encounter figures of horrifying maternal abjection when they meet the darkspawn Broodmother in the Deep Roads. This end-point of corruption is pushed further in the confronting grotesquerie of antagonist Ghilan’nain, a monstrously mutated creator/mother who sculpts the Blight itself. The series introduces the Grey Wardens in the first game, Origins (2009), in which the player character – by something approximating a choice or else kicking and screaming against destiny – must be initiated into their secretive ranks in the opening hours of play. Essentially, they are an order dedicated to keeping the world-threatening peril of the Blight at bay. Spilling up from under the earth, the Blight functions as an eco-gothic threat that can infect and mutate land, water, and people. Sickening crops and poisoning water, the Blight is very much the catastrophic environmental change defined by Smith and Hughes in their seminal discussions of the eco-gothic (5-6). Moreover, the Blight reflects “the non-human ecology rendered uncanny, monstrous” in the changes wrought by its contagious spread (Pinder, para. 1). Thomas Fahy argues that this change, this “preoccupation with hybridity” acts as a recurring motif of the eco-gothic as a mode (3). The fourth game, The Veilguard (2024), is particularly striking in this sense – whole swathes of snowy countryside bubble with blight pools and are encircled by pulsing, blood-heavy tendrils that erupt through the soil, the landscape’s “permeability” made monstrous (Edwards et al., xiv). It is also, crucially, usually dormant – the Grey Wardens, therefore, remain vigilant for decades at a time until they are needed, and until the Blight returns to haunt the landscape. Darkspawn accompany the Blights, monstrous entities that are very much in line with Noël Carroll’s useful framing of “fusion” figures, or monsters that are “amalgams”, that disrupt attempts at categorisation (137). They are recognisably humanoid for the most part, but also consistently skewed or ‘off’ – they move oddly, or appear to have mutated patches of skin or uneven flesh, or they wear armour, but it is positioned strangely on their frames, or their limbs are elongated in a way that feels incorrect. Markedly, they do not speak (with some significant exceptions to be underlined later). While the games broadly draw from classic dark fantasy and Medievalism in equal measure, there is a seam of the Gothic running through them a mile wide, thoroughly exemplified in the darkspawn and the cycles of creeping Blights. Some of the homages are blunt: Ferelden, the nation the player character explores in Origins, is an on-the-nose stand-in for Middle Ages England, complete with wretched weather and a nationalistic love of dogs, along with a long-standing historical feud with neighbouring Orlais, which is France with a few names changed to protect the guilty. Both Dragon Age 2 (2011) and the third game, Dragon Age: Inquisition (2014), do not allow the player to role-play as a Warden. Instead, they tick companion boxes in both. Interestingly, though, the Inquisition’s Warden, Blackwall, is masquerading as a Warden, though he can become one properly later – a secret that the player can guess, though the player character is ignorant of it. They are also the focus of the first game’s expansion, Awakening, and also feature in a host of transmedial connected texts including – but not limited to – short stories, novels such as The Calling, and graphic novels published through Dark Star Digital. In looking at the mutations of the Blight, and the way this is connected inexorably to the Grey Wardens themselves, the focus here will be the first and then the most recent of the games, Origins and Veilguard respectively, since these are the two that force (in Origins’ case) or allow the player to choose (Veilguard) to be a Grey Warden. Fig. 1: The blighted village of D’Meta’s Crossing. Dragon Age: The Veilguard (2024). Screenshot by author. “Join us in the shadows, brothers and sisters”: Grey Warden Secrets As far as monster-hunting secret(ive) societies go, the Grey Wardens beg and borrow most blatantly from Andrzej Sapowski’s Witcher books (and arguably CD Projekt Red’s video game adaptations of the same) and George R.R. Martin’s A Song of Ice and Fire series. Like the former, the Grey Wardens engage with dark magic in order to gain powers that allow them to fight monsters. Similarly, akin to the Night’s Watch and their position as steadfast sentinels, the Grey Wardens vow “in peace, vigilance”, “in death, sacrifice” in their roles as watchers (Origins). A point of difference, however, beyond the initial intertextual referencing at play here, can be found in the manner by which their ranks are initiated. The Joining ritual is not easily survivable – Alistair, one of the companions, notes that “in my Joining, only one of us died. But it was … horrible” (Origins, see fig. 2). Of the two other non-player-characters who are also present at the ritual, Daveth succumbs to death almost immediately after drinking from the chalice. In an unsubtle but decently presented display of the game world’s dark ‘sting in the tail’ narrative approach, the other, Jory, refuses to drink upon witnessing this, and is summarily executed by the senior Warden present in order to keep Warden secrets safe. The games are not shy on the consequences of blighted mutation if the afflicted person does not become a Warden, which is occasionally pointed out as a stopgap cure of sorts. Veilguard’s Antoine and Evka, for instance, are found to a little too frequently accept ‘new recruits’ for the Wardens in the Anderfels in order to help locals who have encountered darkspawn. Instead, they are shunted through the Joining, and then never really expected to become fighters. On the other side of such encounters, the lost dwarf Ruck shies away from torch lights and has lost most of his mind, his skin grey and on the way to rotting; the infected elf Tamlen can ambush the party camp, unrecognisable in his monstrosity, mutated almost entirely into a darkspawn; and an optional player character origin can find themself in need of even more speedy an initiation into the Wardens, since they have had the bad luck to be infected by darkspawn corruption. Fig. 2: The player character (centre of group) as she is about to be initiated through the Joining. Dragon Age: Origins (2009). Screenshot by author. Crucially, the ritual involves the drinking – the ingestion – of darkspawn blood in a gambit that plays almost like a bleak riff on vaccination. If they survive, they will not succumb swiftly to the darkspawn infection if it is passed into their mouth or eyes during combat. This, the player character soon learns, is also the manner by which they will, as a Warden, be able to sense the darkspawn – a connection that both helps but also represents the eventual cost of the Joining. In ingesting darkspawn blood, the process of mutation is slowed – stretched out to maybe thirty years at best – but not halted, and always present. Grey Wardens, therefore, are abject figures that collapse the border between ‘us’ and ‘not us’. The threat of corruption, “death infecting life”, as Julia Kristeva puts it, cannot however in this case be “thrust aside in order to live” (3). Instead, Grey Wardens exist in a corrupted and most Gothic state of mutated becoming. They are adrift amid “indeterminacy and estrangement from one’s own body” (Aldana Reyes 54). So frequently does the Gothic return to the notion of “the coexistence of life and death”, and here, this hybridity is found in the mutating body of the Warden themself (Cavallaro 201). In the game world, this presents in the way Wardens can sense or ‘hear’ darkspawn; in how they, as they grow older, will have terrible, sense-corroding dreams; in how, for some years, they are immune to the blight. It also damns them to their Calling – that point-of-no-return at which individual Wardens find their thoughts overwhelmed by the “song” of the darkspawn, and ritualistically head for the Deep Roads beneath the earth (Origins). While Alistair, optimistic as they come, suggests that this is so the Warden might take out as many darkspawn as they can in combat before being killed in return, it is far easier to read something much darker at work here. In taking themselves into the deep darkness, the hypothetical Grey Warden’s Calling with all its eventual corrupting mutation into a darkspawn necessarily happens out of sight. Interestingly, in her response to Julia Kristeva’s work, Barbara Creed argues for the way social rituals address the abject: that, essentially, “societies both renew their initial contact with the abject element and then exclude that element” (“Monstrous-Feminine” 212). By contrast, the Grey Wardens occupy again that liminal space within which they make that contact and then enshrine it within themselves; no exclusion is permitted. Elizabeth Stang argues for the Grey Wardens here as representing the “normative … order”, as standing in for the heroic and non-monstrous figure opposing the ‘other’, the hero ready to save the land (237). However, this reading softens – or even arguably ignores – the position of the Grey Wardens as part of the horrors they fight. Transformations: The Broodmother under the Surface of Origins The Gothic centrepiece of the Deep Roads sequence in Origins is the player’s encounter with the mutated Broodmother. Deep below the earth, where the darkspawn dwell in horrifying number, the player must work their way through a labyrinth of shadows, corrupted monsters, and the remains of those unlucky enough to have travelled through prior. Catherine Spooner argues that “while Gothic and the grotesque are not reducible to one another, they occupy a similar ... space”, and that is certainly true here (129). The game’s “environmental storytelling” here hinges upon immersion in a Gothic warren that is deliberately designed to be frustrating and horrifying in equal measure – each explored section takes the player and their party further from re-supplying, further from ‘turn-back’ points on the map, further from the daylight far above (Jenkins 122). As Stang discusses in her examination of the Broodmother as the monstrous-feminine, the player’s encounter here is “rife with abject symbolism” (239). Depicted as a monstrously mutated hybrid with multiple pendulous breasts, surrounded by ground that seems more like pulsing flesh, and attended to by darkspawn, the Broodmother is clearly signified as something-that-births. The Broodmother is deeply, confrontingly abject – the mutated maternal figure that is still reproducing, her “generative power” rendered all the more monstrous that it does not seem to need male input (Kristeva 16). As Shohini Chaudhuri frames it, this is a version of the mother of/in abjection, “a parthenogenetic mother, creating all by herself, without need for a father” (95). The lead-up to this encounter is rendered all the more unsettlingly Gothic by Hespith’s explanation of the process of blighted mutation, and how this version of it is horrifyingly different to any form of corruption the player has seen on the surface so far: the men, [the darkspawn] kill… They’re merciful. But the women, they want. They want to touch, to mold, to change until you are filled with them. They took Laryn. They made her eat the others, our friends. She tore off her husband’s face and drank his blood. And while she ate, she grew. She swelled and turned gray and she smelled like them. They remade her in their image. Then she made more of them. Broodmother. (Origins) As Stang notes, the Broodmother is “particularly excessive” in her mutation – this is not the alteration that Tamlen undergoes into a darkspawn ghoul, for instance, in which he still resembles his former self in frame, build, appearance, and so on (240). It is also, strikingly, not the same sort of transformation that Isseya, Veilguard’s impossibly long-lived former Warden mage undergoes (see fig. 3). While Isseya is visibly corrupted, and arguably occupies a similar ‘abject mother’ role – and indeed, she refers to herself as the mother of the griffons she steals – her transformation is one that is designed around a grotesque skeletal appearance that does not immediately infer femininity. Fig. 3: Isseya, the former Warden mage. Dragon Age: The Veilguard (2024). Screenshot by author. By staggering contrast, the Broodmother’s is one that emphasises fleshiness, corporeality, and the reproductive outcome of the transformation (see fig. 4). Stang reads this transformation as inherently problematic in its reliance on the endpoint of the transformation being a corrupted form of maternity (240). In enduring her mutation – in this case, from dwarven explorer to corrupted darkspawn – the Broodmother also loses her voice, and is rendered all the more bestial. Enshrined amid wetness – the floor seems to be unstable, as if it too has become flesh-like – and reached by traversing a dripping tunnel, the Broodmother here is Creed’s “archaic mother” (Monstrous Feminine 18, also cited in Stang 239). This mutated maternity is also of course, horrifyingly, a possible future that awaits female Wardens in pursuit of their Calling. Fig. 4: The Broodmother in combat. Dragon Age: Origins (2009). Screenshot by author. Sculptings: Ghilan’nain’s Bodily Excesses and Experiments in Veilguard A primary antagonist, Ghilan’nain’s design at first calls to mind the monstrosities of cosmic horror. She is tentacled, several metres long (or tall, depending on how she arranges herself), and her upper body comprises two sutured-together torsos (see figs. 5 and 6). Her face, mostly hidden behind a vast mask, gives away only that her eyes are no longer eyes – she appears instead to have boils on one side and teeth beneath the eyelid on the other. Nonetheless, like the Broodmother, she is very much a monster whose monstrosity depends upon her femininity – she speaks in the language of creation, and is known as the “Mother of the Halla” before later revelations unveil her true proclivities and she is referred to as a goddess (Origins). Jane Caputi posits that “patriarchal representations of goddesses are confined to images of fertility, nurturance, and sexual receptivity”, and that once they breach this, they become monsters instead (157). Ghilan’nain can certainly be read this way – she shifts from a caring, deified figure in in-game myth who creates and shepherds the first halla (elven deer, essentially) to a grotesquely mutated tyrant when she emerges in Veilguard. Fig. 5: Ghilan’nain, one of the last remaining Evanuris, lifting up the First Warden and threatening to use his body as fuel for her blight experiments. Dragon Age: The Veilguard (2024). Screenshot by author. She also has one tentacle firmly in the ‘mad scientist’ camp – a grotesque inventor playing with malleable, fleshy toys, with which she creates, reworks, and rebirths darkspawn and dragons with a fondness for vivisection that would impress H.G. Wells’s Doctor Moreau. Tellingly, the player character at one point encounters her works in a “laboratory”, an odd term for the game’s usually more overt Middle Ages setting. It is also here that a past version of herself – a ghost, essentially, a haunting echo – confidently claims, “all that I am belongs to the pursuit of creation. You chose to constrain yourself. I must climb to the heights only understood by gods” (Veilguard). The visual difference in Ghilan’nain herself here is also striking – this past self is missing the tentacles and the added torso, which are heavily implied to have been added through a form of self-surgery. In her work on developing a working definition of the ‘medical gothic’, Laura Kremmel argues that this can be found in “bones and blood”, in “the body’s own viscera”, very much the manner through which Ghilan’nain plans and plots her experiments (21). Like the Gothic itself, Ghilan’nain “knows the body ... knows about physical fragility, about vulnerability”, and her knowledge is turned to monstrous ends (Punter 171). Unlike other blighted monstrous figures, Ghilan’nain maintains control through her ferocious fixation upon experimentation, in which she attempts to turn the mutating presence of the blight into a form of scientific apparatus. Veilguard locates this threat – control over mutation and over the blight – as disorientingly shocking. Following a disastrous encounter with Ghilan’nain at the Grey Wardens’s fortress (see fig. 6), a non-Warden companion points out angrily that “the blight runs in [Wardens’] veins … the same blight that Ghilan’nain commands so effortlessly” (Veilguard 2024). Most strikingly, unlike the Broodmother, however – heretofore Dragon Age’s most pre-eminent female monster – Ghilan’nain is unconstrained, in control of her faculties, creator, not part of the created. For her, the abject filth and flesh of the blight is a tool to be used, rather than something under or through which she is imprisoned. Fig. 6: Ghilan’nain at Weisshaupt, the fortress headquarters of the Grey Wardens. Dragon Age: The Veilguard (2024). Screenshot by author. “In Death, Sacrifice”: Some Final Thoughts Dragon Age games have something of a reputation for tragic endings – or at least, for the possibility of a tragic sting in the tail. In Origins, for instance, it is surprisingly easy to end up with a conclusion that shows the player’s Grey Warden (or Alistair, via a slightly different set of circumstances and choices) stepping wholeheartedly into the “in death, sacrifice” part of their order’s motto. Tanya Krzywinska suggests that many Gothic games waver in their denouement – that what might be termed the game problem of needing a positive or uplifting “winning solution” for the player can smooth out the genre’s inclinations toward the opposite (16). Strikingly though, even if the player sidesteps or schemes their way out of Origins’ still-infamous “sacrifice” ending, the player’s Grey Warden will still, in the imagined narrative space after the credits roll, go on to become a ghoul, to disintegrate into a darkspawn, to find their Calling deep under the earth. Similarly, for the Grey Warden version of Rook, Veilguard’s player character, if they survive the final showdown in Minrathous, they are similarly doomed beyond the game’s brief epilogue. This is an ending that is strikingly Gothic in its insistent corporeality, since the Gothic is, after all, “experienced in the flesh, in its surfaces and crevices” (Aldana Reyes 50). Both possible Wardens here may also be women, which renders their fate the more troubling, since in Dragon Age’s world, the worst, the most abhorrently different of the darkspawn are or have been women before undergoing mutation. As Marie Mulvey-Roberts observes, “monstrosity has been regarded as quintessential to the construction of femininity” (106). For female Grey Wardens, femininity becomes the devastating lynchpin upon which their eventual descent into monstrosity turns. Subjectivity is ultimately at stake once their Calling begins, and that tenuous balance between ‘us’ and ‘not us’ collapses. As Elizabeth Gross argues, the abject – in this case the ingested darkspawn blood – must be kept “at a distance in order [that the subject] define itself as a subject” (87). For the Grey Wardens, the becoming will eventually, inexorably become the became, as the mutating corruption pulls them across to the monstrous side of the binary. References Aldana Reyes, Xavier. Body Gothic: Corporeal Transgression in Contemporary Literature and Horror Film. Cardiff: U of Wales P, 2014. Caputi, Jane. Goddesses and Monsters: Women, Myth, Power and Popular Culture. U of Wisconsin P, 2004. Carroll, Noël. “Fantastic Biologies and the Structures of Horrific Imagery.” The Monster Theory Reader. Ed. Jeffrey Weinstock. U of Minnesota P, 2020. Cavallaro, Dani. The Gothic Vision: Three Centuries of Horror, Terror and Fear. London: Continuum, 2002. Creed, Barbara. “The Monstrous-Feminine: An Imaginary Abjection.” The Monster Theory Reader. Ed. Jeffrey Weinstock. U of Minnesota P, 2020. ———. 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Romantic Medicine and the Gothic Imagination : Morbid Anatomies. Cardiff: U of Wales P, 2022. Kristeva, Julia. Powers of Horror: An Essay on Abjection. Trans. Leon S Roudiez. Columbia UP, 1982. Kryzwinska, Tanya. “The Gamification of Gothic Co-ordinates in Videogames.” Revenant 1.1 (2015). Mulvey-Roberts, Marie. “The Female Gothic Body.” Women and the Gothic: An Edinburgh Companion. Eds. Avril Horner and Sue Zlosnik. Edinburgh UP, 2016. Pinder, Morgan. “‘This mountain belongs to the Wendigo’: The Monstrous Victim in the Wilderness in Horror Video Games.” Gothic Nature (Jan. 2021). Punter, David. Literature of Terror: Volume II: The Modern Gothic. Edinburgh: Pearson Education, 1996. Smith, Andrew, and William Hughes, eds. EcoGothic. Manchester: Manchester UP, 2015. Spooner, Catherine. “Unsettling Feminism: the Savagery of Gothic.” Gothic and Theory: An Edinburgh Companion. Eds. Jerrold Hogle and Robert Miles. Edinburgh UP, 2019. Stang, Sarah. “The Broodmother as Monstrous-Feminine – Abject Maternity in Video Games.” Nordlit 42 (2019): 233-55.