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Brijesh, Pare, B. Jonnalagadda S., S. Tomar Hitendra, Singh Pardeep, and W. Bhagwat V. "Photodegradation of Safranine-O dye using visible irradiation and aqueous suspension of ZnO in a slurry batch reactor." Journal of India Chemical Society Vol. 87, Nov 2010 (2010): 1359–67. https://doi.org/10.5281/zenodo.5805748.
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Laboratory of Photocatalysis, Govt. Madhav Science College, Vikram University, Ujjain-456 010, Madhya Pradesh, India <em>E-mail:</em> brijeshpare2009@gmail.com, pardeepchem@gmail.com School of Chemistry, University of KwaZulu-Natal, Durban, South Africa 'School of Studies in Chemistry, Vikram University, Ujjain-456 010, Madhya Pradesh, India <em>Manuscript received 22 March 2010, accepted 5 May 2010</em> A detailed investigation of photocatalytic degradation of Safranine-O dye has been carried out in the presence of aqueous suspension of ZnO Irradiated with visible light in a batch reactor. It was observed that photocatalytic degradation of Safranine-O obeys pseudo-first order reaction according to Langmuir-Hinselwood model. The effects of various probable influencing factors viz. irradiation time, initial dye concentration, catalyst loading, light intensity, presence of H<sub>2</sub>O<sub>2</sub>, K<sub>2</sub>S<sub>2</sub>O<sub>8</sub>, Na<sub>2</sub>CO<sub>3</sub> and NaCI on the degradation efficiency were systematically studied. The decolourization and extent of degradation of the dye were determined by UV-Visible spectrophotometer and COD reflux method respectively. COD value approximately dropped to zero with the degrading efficiency being about 100% after 3 h. The complete mineralization was confirmed by COD analysis and probable mechanism for the degradation of Safranine-O is proposed.
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Füllemann, P., T. Jörimann, E. D. Bella, M. Stoddart, R. Matthys, and S. Verrier. "IN VITRO 3D STUDY OF THE EFFECT OF UNIAXIAL LOADING ON NAÏVE MSC DIFFERENTIATION FATE." Orthopaedic Proceedings 106-B, SUPP_2 (2024): 135. http://dx.doi.org/10.1302/1358-992x.2024.2.135.
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Bone healing outcome is highly dependent on the initial mechanical fracture environment [1]. In vivo, direct bone healing requires absolute stability and an interfragmentary strain (IFS) below 2% [2]. In the majority of cases, however, endochondral ossification is engaged where frequency and amplitude of IFS are key factors. Still, at the cellular level, the influence of those parameters remains unknown. Understanding the regulation of naïve hMSC differentiation is essential for developing effective bone healing strategies.Human bone-marrow-derived MSC (KEK-ZH-NR: 2010–0444/0) were embedded in 8% gelatin methacryol. Samples (5mm Ø x 4mm) were subjected to 0, 10 and 30% compressive strain (5sec compression, 2hrs pause sequence for 14 days) using a multi-well uniaxial bioreactor (RISystem) and in presence of chondro-permissive medium (CP, DMEM HG, 1% NEAA, 10 µM ITS, 50 µg/mL ascorbic acid, and 100 mM Dex). Cell differentiation was assessed by qRT-PCR and histo-/immunohistology staining. Experiments were repeated 5 times with cells from 5 donors in duplicate. ANOVA with Tukey post-hoc correction or Kurskal-Wallis test with Dunn's correction was used.Data showed a strong upregulation of hypertrophic related genes COMP, MMP13 and Type 10 collagen upon stimulation when compared to chondrogenic SOX9, ACAN, Type 2 collagen or to osteoblastic related genes Type 1 Collagen, Runx2. When compared to chondrogenic control medium, cells in CP with or without stimulation showed low proteoglycan synthesis as shown by Safranine-O-green staining. In addition, the cells were significantly larger in 10% and 30% strain compared to control medium with 0% strain. Type 1 and 10 collagens immunostaining showed stronger Coll 10 expression in the samples subjected to strain compared to control.Uniaxial deformation seems to mainly promote hypertrophic-like chondrocyte differentiation of MSC. Osteogenic or potentially late hypertrophic related genes are also induced by strain.Acknowledgments: Funded by the AO Foundation, StrainBot sponsored by RISystemAG & PERRENS 101 GmbH
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Freeman, Brian C., and Gwyn A. Beattie. "Bacterial Growth Restriction During Host Resistance to Pseudomonas syringae Is Associated with Leaf Water Loss and Localized Cessation of Vascular Activity in Arabidopsis thaliana." Molecular Plant-Microbe Interactions® 22, no. 7 (2009): 857–67. http://dx.doi.org/10.1094/mpmi-22-7-0857.
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The physiological mechanisms by which plants limit the growth of bacterial pathogens during gene-for-gene resistance are poorly understood. We characterized early events in the Arabidopsis thaliana–Pseudomonas syringae pathosystem to identify physiological changes for which the kinetics are consistent with bacterial growth restriction. Using a safranine-O dye solution to detect vascular activity, we demonstrated that A. thaliana Col-0 resistance to P. syringae pv. tomato DC3000 cells expressing avrRpm1 involved virtually complete cessation of vascular water movement into the infection site within only 3 h postinoculation (hpi), under the conditions tested. This vascular restriction preceded or was simultaneous with precipitous decreases in photosynthesis, stomatal conductance, and leaf transpiration, with the latter two remaining at detectable levels. Microscopic plant cell death was detected as early as 2 hpi. Interestingly, suppression of bacterial growth during AvrRpm1-mediated resistance was eliminated by physically blocking leaf water loss through the stomata without altering plant cell death and was nearly eliminated by incubating plants at high relative humidity. The majority of the population growth benefit from blocking leaf water loss occurred early after inoculation, i.e., between 4 and 8 hpi. Collectively, these results support a model in which A. thaliana suppresses P. syringae growth during gene-for-gene resistance, at least in part, by coupling restricted vascular flow to the infection site with water loss through partially open stomata; that is, the plants effectively starve the invading bacteria for water.
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Du, Guoqing, Yi Song, Lei Wei, et al. "Osthole Inhibits Proliferation and Induces Catabolism in Rat Chondrocytes and Cartilage Tissue." Cellular Physiology and Biochemistry 36, no. 6 (2015): 2480–93. http://dx.doi.org/10.1159/000430208.
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Background/Aims: Cartilage destruction is thought to be the major mediator of osteoarthritis. Recent studies suggest that inhibition of subchrondral bone loss by anti-osteoporosis (OP) drug can protect cartilige erosion. Osthole, as a promising agent for treating osteoporosis, may show potential in treating osteoarthritis. The purpose of this study was to investigate whether Osthole affects the proliferation and catabolism of rat chondrocytes, and the degeneration of cartilage explants. Methods: Rat chondrocytes were treated with Osthole (0 μM, 6.25 μM, 12.5 μM, and 25 μM) with or without IL1-β (10ng/ml) for 24 hours. The expression levels of type II collagen and MMP13 were detected by western Blot. Marker genes for chondrocytes (A-can and Sox9), matrix metalloproteinases (MMPs), aggrecanases (ADAMTS5) and genes implicated in extracellular matrix catabolism were evaluated by qPCR. Cell proliferation was assessed by measuring proliferating cell nuclear antigen (PCNA) expression and fluorescence activated cell sorter. Wnt7b/β-catenin signaling was also investigated. Cartilage explants from two-week old SD rats were cultured with IL-1β, Osthole and Osthole plus IL-1β for four days and glycosaminoglycan (GAG) synthesis was assessed with toluidine blue staining and Safranine O/Fast Green FCF staining, collagen type II expression was detected by immunofuorescence. Results: Osthole reduced expression of chondrocyte markers and increased expression of MMP13, ADAMTS5 and MMP9 in a dose-dependent manner. Catabolic gene expression levels were further improved by Osthole plus IL-1β. Osthole inhibited chondrocyte proliferation. GAG synthesis and type II collagen were decreased in both the IL-1β groups and the Osthole groups, and significantly reduced by Osthole plus IL-1β. Conclusions: Our data suggested that Osthole increases the catabolism of rat chondrocytes and cartilage explants, this effect might be mediated through inhibiting Wnt7b/β-catenin pathway.
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Salsabila, Alya, and Gilang Nugraha. "Differences In Storage Time In Safranin Dye On Counting The Number Of Platbocytes By Rees Ecker Method." Jurnal Analis Medika Biosains (JAMBS) 11, no. 1 (2024): 60. http://dx.doi.org/10.32807/jambs.v11i1.347.
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Platelet count examination is important to assess hemostasis function. One of the commonly used manual methods is the Rees Ecker method. However, during the examination, impurities were found that were difficult to distinguish from platelets. Therefore, this study aimed to determine the difference in platelet count results using the Rees Ecker method against storage time with safranin dye. This study used an experimental method on 10 normal blood samples. Platelets were counted with an Improved Neubeauer hemocytometer using safranin dye stored for 0 days, 1 day, 7 days, 14 days, and then compared with the platelet count results using Rees Ecker as a control solution. The Mann Whitney Test results showed no significant difference between the control group and the safranin group in terms of platelet count at 0 days (sig. 0.850), 1 day (sig. 0.820), 7 days (sig. 0.130), and 14 days (sig. 0.121) of storage. Safranin is safe and effective for counting platelets using the Rees Ecker method, as shown by the stability test results for 14 days.
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Encinas, Maria V., Carlos M. Previtali, Marcelo H. Gehlen, and Miguel G. Neumann. "The interaction of aliphatic amines with safranine T in aqueous solution." Journal of Photochemistry and Photobiology A: Chemistry 94, no. 2-3 (1996): 237–41. http://dx.doi.org/10.1016/1010-6030(95)04218-0.
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Rubio-Barroso, S., V. Rodriguez-Gamonal, and L. M. Polo-Diez. "Fluorimetric determination of anionic surfactants by extraction as safranine-T ion-pairs." Analytica Chimica Acta 206 (1988): 357–61. http://dx.doi.org/10.1016/s0003-2670(00)80857-0.
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Maity, S., A. Haldar, and N. B. Manik. "Effect of plasticizer on safranine-T-dye-based solid-state photo electrochemical cell." Ionics 14, no. 6 (2008): 549–54. http://dx.doi.org/10.1007/s11581-008-0217-0.
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Krahmer, R. L., J. J. Morrell, and A. Choi. "Double-Staining to Improve Visualisation of Wood Decay Hyphae in Wood Sections." IAWA Journal 7, no. 2 (1986): 165–67. http://dx.doi.org/10.1163/22941932-90000981.
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Combining fluorescent-labeled wheat germ agglutinin (a chitin-specific lectin) with conventional histological stains offers a simple, efficient method for studying fungal hyphae in deteriorating wood. Cell walls stain dark red with safranin 0, providing excellent contrast for the green-fluorescing hyphae. Staining sections with brilliant vital red markedly enhances the visibility of fungal bore holes.
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Bhattacharya, Subhash C., Haritaran Das, and Satya P. Moulik. "Quenching fluorescence of Safranine T by inorganic ions in aqueous and non-ionic micellar media." Journal of Photochemistry and Photobiology A: Chemistry 84, no. 1 (1994): 39–44. http://dx.doi.org/10.1016/1010-6030(94)03843-0.
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Herrera Millar, Valentina Rafaela, Laura Mangiavini, Umberto Polito, et al. "Hypoxia as a Stimulus for the Maturation of Meniscal Cells: Highway to Novel Tissue Engineering Strategies?" International Journal of Molecular Sciences 22, no. 13 (2021): 6905. http://dx.doi.org/10.3390/ijms22136905.
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The meniscus possesses low self-healing properties. A perfect regenerative technique for this tissue has not yet been developed. This work aims to evaluate the role of hypoxia in meniscal development in vitro. Menisci from neonatal pigs (day 0) were harvested and cultured under two different atmospheric conditions: hypoxia (1% O2) and normoxia (21% O2) for up to 14 days. Samples were analysed at 0, 7 and 14 days by histochemical (Safranin-O staining), immunofluorescence and RT-PCR (in both methods for SOX-9, HIF-1α, collagen I and II), and biochemical (DNA, GAGs, DNA/GAGs ratio) techniques to record any possible differences in the maturation of meniscal cells. Safranin-O staining showed increments in matrix deposition and round-shape “fibro-chondrocytic” cells in hypoxia-cultured menisci compared with controls under normal atmospheric conditions. The same maturation shifting was observed by immunofluorescence and RT-PCR analysis: SOX-9 and collagen II increased from day zero up to 14 days under a hypoxic environment. An increment of DNA/GAGs ratio typical of mature meniscal tissue (characterized by fewer cells and more GAGs) was observed by biochemical analysis. This study shows that hypoxia can be considered as a booster to achieve meniscal cell maturation, and opens new opportunities in the field of meniscus tissue engineering.
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Alibegović, Armin, Nejc Umek, Luka Pušnik, and Inigo Zubiavrre Martinez. "Comparison of the Visual Scoring Method and Semi-Automatic Image Analysis for Evaluating Staining Intensity of Human Cartilage Sections." Image Analysis and Stereology 43, no. 2 (2024): 131–37. http://dx.doi.org/10.5566/ias.3171.
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Accurate estimation of postmortem interval (PMI) is crucial in forensic medicine. The hyaline cartilage, being predominantly composed of a dense extracellular matrix and partly resistant to factors influencing protein degradation, can be utilized for analyzing PMI intervals. Various staining methods are available for cartilage staining for PMI evaluation; however, the conventional visual scoring method for assessing staining intensity is susceptible to evaluator bias. This study compared the visual scoring method with a modified Bern score with semi-automatic image analysis. The cartilage samples were obtained from three cadavers with known time of death. Forty-five histological slices were prepared and stained using Alcian blue, Safranin-O with Fast green, Safranin-O without Fast green, Masson trichrome, and Sirius red. Ten evaluators visually scored each sample on a scale of 0 to 3. A semi-automatic analysis was conducted on the same images using the deconvolution plugin of the ImageJ software. Linear regression was used to assess the correlation between the mean grey value and the mean Bern score from all evaluators. The results showed strong correlations across all evaluated staining techniques, with Masson trichrome staining exhibiting the highest correlation. Accordingly, semi-automatic image analysis can be a suitable replacement for the visual scoring method, particularly when no procedural artifacts are present.
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Shoemaker, R. Stuart, Alicia L. Bertone, George S. Martin, et al. "Effects of intra-articular administration of methylprednisolone acetate on normal articular cartilage and on healing of experimentally induced osteochondral defects in horses." American Journal of Veterinary Research 53, no. 8 (1992): 1446–53. http://dx.doi.org/10.2460/ajvr.1992.53.08.1446.
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Summary The effects of intra-articular administration of methylprednisolone acetate (mpa) on the healing of full-thickness osteochondral defects and on normal cartilage were evaluated in 8 horses. In group-1 horses (n = 4), a 1-cm-diameter, full-thickness defect was created bilaterally in the articular cartilage on the dorsal distal surface of the radial carpal bone. Cartilage defects were not created in group-2 horses (n = 4). One middle carpal joint was randomly selected in each horse (groups 1 and 2), and treated with an intra-articular injection of 100 mg of mpa, once a week for 4 treatments. Injections began 1 week after surgery in group-1 horses. The contralateral middle carpal joint received intra-articular injections of an equivalent volume of 0.9% sodium chloride solution (scs), and served as a control. Horses were evaluated for 16 weeks, then were euthanatized, and the middle carpal joints were examined and photographed. Synovial and articular cartilage specimens were obtained for histologic and histochemical evaluation. Gross morphometric evaluation of the healing defects in group-1 horses revealed that 48.6% of the defect in control joints and 0% of the defect in mpa-treated joints was resurfaced with a smooth, white tissue, histologically confirmed as fibrocartilage. This replacement tissue was a firmly attached fibrocartilage in control joints and a thin fibrous tissue in mpa-treated joints. The articular cartilage in joints treated with mpa had morphologic changes, including chondrocyte cluster formation, loss of palisading architecture, and cellular necrosis in both groups of horses. Histochemical (safranin-0) staining intensity was reduced significantly (P < 0.05) in all layers of articular cartilage in mpa-treated joints in groups 1 and 2. In the replacement tissue, intense safranin-0 staining was found only in the chondrocyte clusters deep in the tissue of control joints, confirming fibrocartilage repair. Intra-articular administration of mpa in this dosing regimen thus induced degenerative changes in normal articular cartilage and resulted in histomorphologic changes in the repair of full-thickness articular osteochondral defects in horses.
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Nandi, Susantamay, and Subhash C. Bhattacharya. "Alkanol effects on fluorescence quenching of Safranine T by thiourea in reverse micellar medium of Aerosol–O–T in heptane." Colloids and Surfaces A: Physicochemical and Engineering Aspects 186, no. 3 (2001): 179–87. http://dx.doi.org/10.1016/s0927-7757(00)00822-0.
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KELLER, SCOTT, JOSEPH MARCY, BARBARA BLAKISTONE, CAMERON HACKNEY, W. HANS CARTER, and GEORGE LACY. "Application of Fluid Modeling To Determine Threshold Leak Size for Liquid Foods." Journal of Food Protection 66, no. 7 (2003): 1260–68. http://dx.doi.org/10.4315/0362-028x-66.7.1260.
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In this study, the mechanism by which a package defect converts to a leaker was examined in an effort to develop a relationship between threshold leak size and loss of package sterility. The threshold leak size is the hole size at which the onset of leakage occurs. The threshold pressure is the pressure required to initiate a leak. Leak initiation was studied in terms of the interaction between three components: liquid attributes of liquid food products, defect size, and pressures required to initiate liquid flow. Liquid surface tension, viscosity, and density values were obtained for 16 liquids. The imposed pressures required to initiate flow through microtubes with interior diameters of 0, 2, 5, 7, 10, 20, and 50 μm were measured with the use of 63 test cells filled with safranin red dye, tryptic soy broth, and distilled water with surface tensions of 18.69, 44.09, and 64.67 mN/m, respectively. Significant differences (P &lt; 0.05) between threshold pressures observed for safranin red dye, tryptic soy broth, and distilled water were found. Liquids with low surface tensions, such as safranin red dye, required significantly lower threshold imposed pressures than did liquids with high surface tensions, such as distilled water (P &lt; 0.05). An equation to quantify the relationship between liquid surface tension, threshold imposed pressure, and defect size was developed. Threshold pressures observed were not significantly different (P &gt; 0.05) from those predicted by the equation. Imposed pressures and vacuums generated within packages during random vibration and sweep resonance tests were measured for brick-style aseptic packages (250 ml), metal cans (76.2 by 114.3 mm [425 ml]), 1-qt gable-top packages (946 ml), 0.5-gal gable-top packages (1.89 liters), and 1-gal milk jugs (4.25 liters). Significant differences between packages were found with respect to observed generated pressures during vibration testing (P &lt; 0.05). An equation to calculate threshold size on the basis of liquid surface tension and imposed pressure was established.
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Bhattacharya, Subhash C., Susantamay Nandi, and Satya P. Moulik. "Visible and fluorescence spectral studies on the interaction of Safranine T with reverse micelles of Triton X-100 and Tweens in chloroform." Journal of Photochemistry and Photobiology A: Chemistry 97, no. 1-2 (1996): 57–63. http://dx.doi.org/10.1016/1010-6030(96)04308-0.
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GÜMÜŞ, Dilek. "ORGANİK ATIKLARDAN ÜRETİLEN KOMPOZİT BİR MODİFİYE BİYOKÖMÜR KULLANILARAK SULU ÇÖZELTİDEN SAFRANİN T GİDERİMİ." Kahramanmaraş Sütçü İmam Üniversitesi Mühendislik Bilimleri Dergisi 25, no. 3 (2022): 237–48. http://dx.doi.org/10.17780/ksujes.1097965.
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Modern yaşamın ilgi çekici unsurlarından biri de renktir. Başta tekstil endüstrileri olmak üzere bir çok endüstri, farklı konsantrasyonlarda çeşitli boyalar içeren yüksek miktarda renkli atık su üretmektedir. Sağlık ve ekolojik kaygılara yol açan sentetik boyaların alıcı ortama verilmeden önce atıksulardan uzaklaştırılması gerekmektedir. Düşük maliyetli adsorbanlar elde edebilmek için atık malzemelerin kullanımı, atıksu arıtma maliyetlerinin azaltılmasına ve çevrenin korunmasına katkıda bulunduğu için araştırmalara konu olmaktadır. Bu çalışmada, kolayca temin edilebilen ve toksik olmayan organik üretilen aktive edilmiş kompozit bir biyokömür hazırlanarak Safranin T boyasının gideriminde adsorbent olarak kullanılmıştır. Kesikli sistemde gerçekleştirilen deneylerde adsorban miktarı (0,1-1 g/L), boya konsantrasyonu (10-50 mg/L), pH (5-9) ve temas süresi (0-360 dk) gibi en temel parametreler incelenmiştir. Dört farklı İzoterm ve dört farklı kinetik tartışılmıştır. Elde edilen verilerle Langmuir izoterm modeli ve sözde ikinci derece kinetik model daha iyi uyum sağlamıştır.
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Trotter, Gayle W., C. Wayne McIlwraith, John V. Yovich, Robert W. Norrdin, Robert H. Wrigley, and C. H. Lamar. "Effects of intra-articular administration of methylprednisolone acetate on normal equine articular cartilage." American Journal of Veterinary Research 52, no. 1 (1991): 83–87. http://dx.doi.org/10.2460/ajvr.1991.52.01.83.
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SUMMARY The effects of the corticosteroid 6-α-methylprednisolone acetate on normal equine articular cartilage were evaluated, using the middle carpal joint in 4 clinically normal young horses. One middle carpal joint of each horse was injected 3 times with 100 mg of 6-α-methylprednisolone acetate, at 14-day intervals. The opposite middle carpal joint (control) was injected with 2.5 ml of lactated Ringer solution at the same intervals. Effects were studied until 8 weeks after the first injection. Evaluation included clinical and radiographic examination, and gross, microscopic, and biochemical evaluation of joint tissues. Horses remained clinically normal during the study, and significant radiographic changes were not observed. Safranin-0 matrix staining intensity and uronic acid content were significantly (P < 0.05) lower and hydroxyproline content was significantly (P < 0.05) higher in articular cartilage of corticosteroid-injected joints vs control joints.
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Prabowo, Ian, and Diah Rachmawati. "RESPONS FISIOLOGIS DAN ANATOMI AKAR TANAMAN BAYAM (Amaranthus tricolor L.) TERHADAP CEKAMAN NaCl." Jurnal Penelitian Saintek 25, no. 1 (2020): 36–43. http://dx.doi.org/10.21831/jps.v25i1.27357.
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Tujuan dari penelitian ini untuk mengetahui pengaruh cekaman NaCl terhadap pertumbuhan dan anatomi akar tanaman bayam serta mengetahui konsentrasi NaCl yang dapat menghambat pertumbuhan tanaman bayam. Penelitian ini digunakan perlakuan NaCl sebanyak 0, 200, 400, 600 dan 800 mM pada tanaman bayam (Amaranthus tricolor L.), Media tanam berupa campuran tanah dan pupuk kandang dan alkohol, safranin, aseton. Alat yang digunakan adalah med line, cawan poerslen, spektrofotometer dan mikroskop. Parameter yang diukur meliputi tinggi tanaman, jumlah daun, warna daun, panjang akar, jumlah akar, berat basah, berat kering, kadar klorofil, tebal epidermis akar, tebal korteks akar dan diameter stele akar. Data dianalisis dengan uji ANOVA dilanjutkan dengan DMRT taraf kepercayaan 95% menggunakan program SPSS 15. Hasil yang diperoleh menunjukkan penambahan NaCl menyebabkan penurunan pertumbuhan tanaman bayam meliputi tinggi tanaman, jumlah daun, kadar klorofil, rasio tajuk dibanding akar dan menurunkan diameter stele akar. Pertumbuhan tanaman menurun seiring peningkatan konsentrasi NaCl karena NaCl menyebabkan cekaman osmotik yang menghambat penyerapan air dan unsur hara yang diperlukan tanaman untuk proses metabolisme.PHYSIOLOGICAL RESPONSE AND ANATOMY OF ROOTY PLANT [ Amaranthus tricolor L.] AGAINST NaClThe study was aimed at determining the effect of NaCl stress on the growth and anatomy of spinach roots and the concentration of NaCl which can inhibit the growth of spinach plants. This study used 0, 200, 400, 600 and 800 mM NaCl treatments on spinach (Amaranthus tricolor L.), planting media in the form of a mixture of soil and manure and alcohol, safranin, acetone. Med line, poerslen cup, spectrophotometer, and microscope were used in this study. The parameters measured plant height, number of leaves, leaf color, root length, number of roots, wet weight, dry weight, chlorophyll content, root epidermis thickness, root cortex thickness, and root stele diameter. The collected data then were analyzed by ANOVA test followed by DMRT 95% confidence level using the SPSS 15 program. The results obtained showed that the addition of NaCl caused a decrease in spinach plant growth including plant height, the number of leaves, chlorophyll content, the ratio of the crown to root and decreased diameter of root stele. Plant growth decreases with increasing NaCl concentration since NaCl causes osmotic stress. This stress inhibits the absorption of water and nutrients needed by plants for metabolic processes.
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Ningrum, Eka Fitriana Candra, Ikhsanudin Nur Rosyidi, Rizka Riliant Puspasari та Endang Semiarti. "Perkembangan Awal Protocorm Anggrek Phalaenopsis amabilis secara In Vitro setelah Penambahan Zat Pengatur Tumbuh α-Naphtaleneacetic Acid dan Thidiazuron". Biosfera 34, № 1 (2017): 9. http://dx.doi.org/10.20884/1.mib.2017.34.1.393.
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Phalaenopsis amabilis (L.) Blume merupakan salah satu anggrek alam dengan nilai komersial yang tinggi. Keberadaannya di alam semakin sukar dijumpai akibat eksploitasi berlebihan dan kerusakan hutan. Selain itu budidaya anggrek ini cukup sulit dilakukan karena biji bersifat mikroskopis dan tidak memiliki endosperm, sehingga perlu dilakukan kultur in vitro. Dalam budidaya secara in vitro diperlukan medium pendukung pertumbuhan anggrek secara optimal misalnya dengan penambahan Zat Pengatur Tumbuh (ZPT) dari golongan auksin dan sitokinin. Penelitian ini bertujuan untuk mengetahui pengaruh penambahan ZPT tersebut terhadap perkembangan protokorm (pertumbuhan embrio anggrek) P. amabilis secara in vitro dari aspek morfologi dan anatomi. Penelitian dilakukan di Laboratorium Bioteknologi dan Laboratorium Struktur Perkembangan Tumbuhan. Digunakan protokorm anggrek P. amabilis berumur 2 bulan. Protokorm disubkultur pada medium NP dengan kombinasi ZPT NAA dan TDZ pada berbagai konsentrasi dan diinkubasi pada ruangan dengan kelembaban tinggi, suhu 250C. Kemudian diamati protokorm anggrek selama 2 minggu yang mencakup persentase pertumbuhan, persentase protokorm yang membentuk tunas dan absorbing hair. Pembuatan preparat anatomi dilakukan dengan mengambil protokorm setiap minggunya, disimpan dalam larutan alkohol 70% dan dibuat preparat anatomi menggunakan metode Parafin dengan pewarnaan safranin. Hasil penelitian menunjukkan pada penambahan NAA dan TDZ pertumbuhan protokorm mencapai 100%. Persentase protokorm yang membentuk tunas berturut turut (0:0, 0.2:0, 0:0.5, 0.2:0.5, 0:1, dan 0.2:1) ppm adalah 24%, 22%, 32%, 40%, 40% dan 38% sedangkan untuk pengamatan protokorm yang membentuk absorbing hair sebesar 10%, 18%, 14%, 24%, 16%, dan 0%. Untuk diameter sel berukuran 39.2±0.47µm, 44.4±0.97µm, 39.08±0.5µm, 38.83±0.2µm,39.5±0.39µm dan 39.75±0.28µm. Sehingga dapat disimpulkan medium yang paling optimal dalam pertumbuhan anggrek P. amabilis secara in vitro adalah medium NP dengan kombinasi NAA dan TDZ (0.2 : 0.5) ppm.Kata kunci : Anatomi, Kultur In Vitro, Morfologi, Phalaenopsis amabilis
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Roosendaal, Goris, Simon Mastbergen, Katja Coeleveld, et al. "Deferasirox limits cartilage damage following haemarthrosis in haemophilic mice." Thrombosis and Haemostasis 112, no. 11 (2014): 1044–50. http://dx.doi.org/10.1160/th14-01-0029.
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SummaryJoint bleeds in haemophilia result in iron-mediated synovitis and cartilage damage. It was evaluated whether deferasirox, an iron chelator, was able to limit the development of haemophilic synovitis and cartilage damage. Haemophilic mice were randomly assigned to oral treatment with deferasirox (30 mg/kg) or its vehicle (control) (30 mg/kg). Eight weeks after start of treatment, haemarthrosis was induced. After another five weeks of treatment, blood-induced synovitis and cartilage damage were determined. Treatment with deferasirox resulted in a statistically significant (p< 0.01) decrease in plasma ferritin levels as compared to the control group (823 ng/ml ± 56 and 1220 ng/ml ±114, respectively). Signs of haemophilic synovitis, as assessed by the Valentino score [range 0 (normal) – 10 (most affected)], were not different (p=0.52) when comparing the control group with the deferasirox group. However, deferasirox treatment resulted in a statistically significant (p< 0.01) reduction in cartilage damage, as assessed by the loss in Safranin O staining [range 0 (normal) – 6 (most affected)], when comparing the deferasirox group with the control group: score 2 (65.4 % vs 4.2 %), score 3 (26.9 % vs 4.2 %), score 4 (7.7 % vs 20.8 %), score 5 (0 % vs 54.2 %), and score 6 (0 % vs 16.7 %). Treatment with deferasirox limits cartilage damage following the induction of a haemarthrosis in haemophilic mice. This study demonstrates the role of iron in blood-induced cartilage damage. Moreover, these data indicate that iron chelation may be a potential prevention option to limit the development of haemophilic arthropathy.
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Gilead, L., O. Bibi, and E. Razin. "Fibroblasts induce heparin synthesis in chondroitin sulfate E containing human bone marrow-derived mast cells." Blood 76, no. 6 (1990): 1188–95. http://dx.doi.org/10.1182/blood.v76.6.1188.1188.
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Abstract Human bone marrow-derived mast cells (hBMMCs), differentiated in vitro in suspension culture and under the influence of human peripheral blood mononuclear cells conditioned medium (hCM), were tested for their response to recombinant human interleukin-3 (rhIL-3) and for their behavior in different microenvironments. The hBMMCs were incubated in the presence of rhIL-3 and the changes in their proliferation rate were determined. Recombinant hIL-3 induced a more than sixfold increase in 3H-thymidine uptake into the hBMMC DNA in a dose-dependent manner. Human CM used as a control for proliferation response induced a more than eightfold maximal proliferation rate increase. Rabbit anti-rhIL-3 completely inhibited hBMMC 3H-thymidine uptake induced by rhIL-3 and decreased the hCM-induced proliferation by approximately 50%. These hBMMCs were cocultured with four different mytomicin C-treated cell monolayers and assayed for phenotypic changes. After only 2 days in coculture with either embryonic mouse skin-derived fibroblasts (MESFs) or human skin-derived fibroblasts (HSFs), a marked increase in granule number and density was noted on staining with toluidine blue. Mast cells that initially stained alcian blue+/safranin- at day 0 of coculture became alcian blue+/safranin+ during the coculture period. Human BMMC proteoglycan synthesis shifted from approximately 85% chondroitin sulfate E to approximately 60% heparin within 14 to 19 days of coculture with the MESF monolayer and to approximately 50% heparin within 19 days of coculture with the HSF monolayer. None of the above- mentioned changes were noted in cocultures of hBMMCs with 3T3 cell line fibroblast monolayers or in cocultures with bovine vascular endothelium (BVE) cell monolayers. These results demonstrate microenvironmental effects exerted by the MESF and HSF monolayers on IL-3-dependent hBMMCs similar to those reported in the conversion of murine mast cell phenotype.
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Gilead, L., O. Bibi, and E. Razin. "Fibroblasts induce heparin synthesis in chondroitin sulfate E containing human bone marrow-derived mast cells." Blood 76, no. 6 (1990): 1188–95. http://dx.doi.org/10.1182/blood.v76.6.1188.bloodjournal7661188.
Full textAbstract:
Human bone marrow-derived mast cells (hBMMCs), differentiated in vitro in suspension culture and under the influence of human peripheral blood mononuclear cells conditioned medium (hCM), were tested for their response to recombinant human interleukin-3 (rhIL-3) and for their behavior in different microenvironments. The hBMMCs were incubated in the presence of rhIL-3 and the changes in their proliferation rate were determined. Recombinant hIL-3 induced a more than sixfold increase in 3H-thymidine uptake into the hBMMC DNA in a dose-dependent manner. Human CM used as a control for proliferation response induced a more than eightfold maximal proliferation rate increase. Rabbit anti-rhIL-3 completely inhibited hBMMC 3H-thymidine uptake induced by rhIL-3 and decreased the hCM-induced proliferation by approximately 50%. These hBMMCs were cocultured with four different mytomicin C-treated cell monolayers and assayed for phenotypic changes. After only 2 days in coculture with either embryonic mouse skin-derived fibroblasts (MESFs) or human skin-derived fibroblasts (HSFs), a marked increase in granule number and density was noted on staining with toluidine blue. Mast cells that initially stained alcian blue+/safranin- at day 0 of coculture became alcian blue+/safranin+ during the coculture period. Human BMMC proteoglycan synthesis shifted from approximately 85% chondroitin sulfate E to approximately 60% heparin within 14 to 19 days of coculture with the MESF monolayer and to approximately 50% heparin within 19 days of coculture with the HSF monolayer. None of the above- mentioned changes were noted in cocultures of hBMMCs with 3T3 cell line fibroblast monolayers or in cocultures with bovine vascular endothelium (BVE) cell monolayers. These results demonstrate microenvironmental effects exerted by the MESF and HSF monolayers on IL-3-dependent hBMMCs similar to those reported in the conversion of murine mast cell phenotype.
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Abbaz, Amina, Sihem Arris, Gianluca Viscusi, Asma Ayat, Halima Aissaoui, and Yasser Boumezough. "Adsorption of Safranin O Dye by Alginate/Pomegranate Peels Beads: Kinetic, Isotherm and Thermodynamic Studies." Gels 9, no. 11 (2023): 916. http://dx.doi.org/10.3390/gels9110916.
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Water pollution is regarded as a dangerous problem that needs to be resolved right away. This is largely due to the positive correlation between the increase in global population and waste production, especially food waste. Hydrogel beads based on sodium alginate (Alg) and pomegranate fruit peels (PP) were developed for the adsorption of Safranin O dye (SO) in aqueous solutions. The obtained Alg−PP beads were widely characterized. The effects of the contact time (0–180 min), initial concentration (10–300 mg/L), initial pH (2–10), adsorbent dosage (1–40 g/L) and the temperature (293–333 K) were investigated through batch tests. The data proved that the adsorption kinetics of SO reached equilibrium within 30 min and up to 180 min. The dye adsorption is concentration dependent while a slight effect of pH was observed. The adsorption data of SO onto synthesized beads follow the pseudo second-order model. The experimental data fitted very well to Langmuir model with correlation factor of 0.92 which demonstrated the favourable nature of adsorption. The maximum adsorption capacity of Alg−PP could reach 30.769 mg/g at 293 K. Calculation of Gibbs free energy and enthalpy indicated that adsorption of SO onto Alg−PP is spontaneous (negative ΔG) and endothermic (ΔH = 9.30 kJ/mol). Analysis of diffusion and mass transport phenomena were presented. The removal efficiency was found to be 88% at the first cycle and decreased to 71% at the end of the seventh cycle. The reported results revealed that the Alg−PP beads could be used as a novel natural adsorbent for the removal of high concentrated solutions of Safranin O which is a cationic dye from liquid affluents and as future perspective, it can be used to remove various pollutants from wastewater.
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Dubuc, Julia, Melodie Jil Schneider, Valerie Dubuc, et al. "Degradation of Proteoglycans and Collagen in Equine Meniscal Tissues." International Journal of Molecular Sciences 25, no. 12 (2024): 6439. http://dx.doi.org/10.3390/ijms25126439.
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Investigate meniscal extracellular matrix degradation. Equine menisci (n = 34 from 17 horses) were studied. Site-matched sections were cut and scored from three regions (ROIs; n = 102) and stained for histology, proteoglycan (safranin O and fast green), aggrecan, and collagen cleavage (NITEGE, DIPEN, and C1,2C antibodies, respectively). Picrosirius red and second harmonic generation microscopy were performed to investigate collagen ultrastructure. A total of 42 ROIs met the inclusion criteria and were included in the final analysis. The median (range) ROI histological score was 3 (0–9), providing a large spectrum of pathology. The median (range) proteoglycan score was 1 (0–3), representing superficial and central meniscal loss. The median (range) of DIPEN, NITEGE, and C1,2C scores was 1 (0–3), revealing immunostaining of the femoral and tibial surfaces. The proteoglycan scores exhibited significant positive associations with both histologic evaluation (p = 0.03) and DIPEN scores (p = 0.02). Additionally, a robust positive association (p = 0.007) was observed between the two aggrecanolysis indicators, NITEGE and DIPEN scores. A negative association (p = 0.008) was identified between NITEGE and histological scores. The C1,2C scores were not associated with any other scores. Picrosirius red and second harmonic generation microscopy (SHGM) illustrated the loss of the collagen matrix and structure centrally. Proteoglycan and collagen degradation commonly occur superficially in menisci and less frequently centrally. The identification of central meniscal proteoglycan and collagen degradation provides novel insight into central meniscal degeneration. However, further research is needed to elucidate the etiology and sequence of degradative events.
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Smith, H. V., A. McDiarmid, A. L. Smith, A. R. Hinson, and R. A. Gilmour. "An analysis of staining methods for the detection ofCryptosporidiumspp. oocysts in water-related samples." Parasitology 99, no. 3 (1989): 323–27. http://dx.doi.org/10.1017/s0031182000059023.
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SummaryThe ability of five staining techniques, originally developed for the rapid identification ofCryptosporidiumspp. oocysts in faecal samples, to detect oocysts in water and water-related samples was assessed. All the stains used (modified Ziehl Neelsen, auramine-phenol (Lempert), Wright-Giemsa, safranin-methylene blue and FITC-labelled monoclonal antibody) stained oocysts after storage in water for 2 months at 4 °C (71–89 % of control values). Storage of oocysts below 0 °C greatly reduced the staining ability of auramine-phenol. With the exception of oocysts stored in raw and final waters, the histochemical stains proved less useful in detecting oocysts than the monoclonal antibody. Organisms of similar size and shape took up these stains, causing confusion in interpretation. Cold Ziehl Neelsen and the FITC-labelled monoclonal antibody were best at identifying oocysts from a waterborne outbreak. Screening with a fluorescent antibody, followed by confirmation with cold Ziehl Neelsen, where possible, are the currently recommended procedures for the detection of oocysts in water-related samples.
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Dassler, C. L., S. M. Griffey, and P. B. Vasseur. "Histological features of osteoarthritic canine cartilage after prolonged administration of carprofen." Veterinary and Comparative Orthopaedics and Traumatology 16, no. 01 (2003): 32–37. http://dx.doi.org/10.1055/s-0038-1632752.
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SummaryThe objective of this study was to determine whether the non-steroidal anti-inflammatory drug (NSAID) carprofen causes adverse histological changes to cartilage after prolonged administration for treatment of naturally acquired osteoarthritis.Dogs diagnosed with cranial cruciate ligament rupture and secondary osteoarthritis of their stifle joints were divided into two groups: those who had received a standard dose of carprofen for a minimum of four weeks and those who had not received a NSAID for a minimum of one month. Cartilage from the intercondylar notch was obtained when the patients underwent surgical procedures to stabilize the stifle joint. The samples were processed and evaluated for histological differences using haematoxylin and eosin and safranin-O-fast green-iron haematoxylin stains and were graded according to a Mankin scale (graded 0-14) for osteoarthritis.The histopathological findings were variable and included loss of matrix staining, irregularities to the cartilage surface, matrix fragmentation, chondrocyte clustering, matrix degeneration and vascular and synovial invasion (pannus) and were not specific to either group. There was not any significant difference between study groups (p = 0.2721) according to the Wilcoxan rank sum test.
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Bertone, Alicia L., C. Wayne McIlwraith, Robert L. Jones, Robert W. Norrdin, M. Judith Radin, and Jack L. Lebel. "Comparison of various treatments for experimentally induced equine infectious arthritis." American Journal of Veterinary Research 48, no. 3 (1987): 519–29. https://doi.org/10.2460/ajvr.1987.48.03.519.
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SUMMARY To evaluate the effects of 5 treatments on clinical responses, synovial fluid analysis, radiographic changes, bacteriologic culture results of the synovial fluid and synovial membrane, microscopic characteristics of the synovial membrane, and articular cartilage histochemistry, Staphylococcus aureus organisms (1.6 × 106 colony-forming units) were inoculated into the tarsocrural joints of 12 horses (n = 24 joints; 2 joints/horse). Each horse was given phenylbutazone (2 g) orally, every 24 hours, beginning 24 hours after inoculation. Two horses (ie, 4 joints) were not given other treatment (controls; group 1). All other horses (ie, 20 joints) were given a trimethoprim-sulfadiazine combination orally, once daily (30 mg/kg; 8 joints) or twice daily (30 mg/kg q 12 hr; 12 joints). Each of these 20 joints were assigned to 1 of 5 treatment groups (4 joints/group) in a balanced incomplete block design. Group 2 (4 joints) was given only the antibiotics once daily. Twelve joints were treated by through-and-through joint lavage on day 1 (group 3), days 1 and 3 (group 4), or days 1, 3, and 6 (group 5). Joints in group 6 had an arthrotomy performed on day 1, with subsequent lavage via an indwelling drain every 12 hours for 4 days. In groups 3 through 6, 1 joint in each group was treated with antibiotics once daily, and 3 joints were treated with antibiotics twice daily. All horses were clinically assessed each day. Complete blood count was performed on days 3, 6,10, and 21. Before inoculation and on days 0, 1, 3, 6, 10, and 21, synovial fluid specimens were collected and analyzed for color, clarity, total protein concentration, wbc count, differential count, and mucin clot-forming ability. Synovial fluid specimens were cultured bacteriologically before inoculation and on days 0 and 21. Horses in group 1 (controls) were euthanatized before day 6. All other horses were euthanatized on day 21. Tarsocrural joints were opened and examined. Synovial membrane specimens were bacteriologically cultured. Synovial membrane specimens were examined histologically (hemotoxylin and eosin stain) and articular cartilage specimens were (safranin O fast green stain) evaluated histochemicallv. Synovial membrane specimens were histologically graded into 5 categories. Intensity of articular cartilage intercellular staining with safranin 0 was graded for superficial, outer intermediate, inner intermediate, and deep zones. Two-way analysis of variance was performed to evaluate differences among groups and across time for the determinants evaluated. A 1-way analysis of variance was performed to evaluate differences in the peripheral clinicopathologic determinants across time and to evaluate histologic and histochemic differences among groups. To determine significance, the probability of a type-I error was < 0.05. Horses with joints assigned to group 2 (administered antibiotics once daily) had intermittent pyrexia. Differences in other clinical determinants were not detected among treatment groups. All horses had a low pcv, high plasma fibrinogen concentrations, and high peripheral wbc counts between days 3 and 10. The tarsocrural joints did not have radiographic evidence of bone erosion or new bone formation. In joints in which an arthrotomy was performed, less soft tissue swelling was apparent on day 10, compared with other treatment groups (P = 0.1). Synovial fluid was flocculent after day 3 in all groups, except group 6 (arthrotomy). On day 21, joints in group 6 had a higher (P = 0.01) synovial fluid protein concentration than did joints in groups 2 or 3. Differences in synovial fluid, wbc count, differential count, or mucin clot-forming ability were not detected among the groups. At the termination of the study, Staphylococcus aureus was isolated from the synovial fluid and from the synovial membrane specimens of 87% of the joints treated with once-daily administration of antibiotics and of 42% of the joints treated with twice-daily administration of antibiotics (P < 0.05). Joint drainage (groups 3 through 6) did not affect bacteriologic isolation rates. Synovial membrane specimens from joints in which an arthrotomy was performed had significantly less histologic evidence of inflammation (P < 0.05), compared with other treatment groups. Articular cartilage specimens from joints treated with antibiotics once daily and for which joint drainage was not performed (group 2) had less intense intercellular staining with safranin O (ie, greater loss of glycosaminoglycans) than did other treatment groups. Systemic administration of antibiotics (even in low doses, ie, administered once daily) resulted in clinical improvement in appetite, comfort, and other clinical determinants measured, compared with control horses. Treatment with higher doses of antibiotics (ie, administered twice daily) decreased the rate of bactériologie isolation, and, with joint drainage, resulted in less loss of articular cartilage glycosaminoglycans (ie, significantly greater staining intensity with safranin O) than joints treated with antibiotics once daily. Arthrotomy with lavage resulted in less synovitis and less fibrin formation than treatment by through-and-through lavage performed 1 to 3 times. Delayed healing, fibrosis, and excessive granulation tissue complicated healing of all arthrotomy incisions. Rates of bacterial isolation were identical from synovial fluid specimens and from synovial membrane specimens.
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Kincaid, S. A., R. V. Allhands, and G. J. Pijanowski. "Chondrolysis associated with cartilage canals of the epiphyseal cartilage of the distal humerus of growing pigs." American Journal of Veterinary Research 46, no. 3 (1985): 726–32. https://doi.org/10.2460/ajvr.1985.46.03.726.
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SUMMARY The articular-epiphyseal (a-e) cartilage of the distal humeri of 7 pigs weighing 13.1 to 18.2 kg and of 3 pigs weighing 36.4 to 40.9 kg was studied. Frozen samples of a-e cartilage were stained for the presence of reduced nicotinamide adenine dinucleotide dehydrogenase, lactate dehydrogenase, isocitrate dehydrogenase, and uridine diphosphate galactose-4-epimerase. Additional frozen sections and paraffin-processed sections were stained using the Alcian blue-critical electrolyte concentration method, safranin 0-fast green, and hematoxylin and eosin. An area of grossly visible, opaque a-e cartilage of the medial condyle corresponded to regions of chondrolysis of the epiphyseal cartilage. The chondrolytic regions contained chondrocytes that did not stain for enzymes, had reduced staining for proteoglycans in the matrix, and were located at the site where the a-e cartilage increased in thickness. Cartilage canals were associated with the chondrolytic areas. Cartilage canals in both groups of pigs were commonly in various stages of chondrification, some of which were associated with degenerative cartilage. The regions of chondrolysis may indicate sites of biomechanical weakness in the a-e cartilage during the transformation of the epiphyseal cartilage into bone.
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Nisa Dewi Setyawati, Handoko Santoso, and Anak Agung Oka. "UJI KETAHANAN PREPARAT JARINGAN TUMBUHAN MENGGUNAKAN PEWARNA ALAMI BUAH NAGA (Hylocereus polyrhizus) dan KUNYIT (Curcuma domestica Val) DENGAN VARIASI WAKTU SEBAGAI SUMBER BELAJAR BIOLOGI DALAM BENTUK LKPD." EDUBIOLOCK 5, no. 2 (2024): 23–32. https://doi.org/10.24127/edubiolock.v5i2.6495.
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The aims of this study were 1) to determine the effect of time variation on preparation endurance, 2) to determine the best time variation on preparation endurance, 3) to determine whether LKPD (Student Activity Sheets) were appropriate as learning resources at school. This type of research is descriptive qualitative experimental research. This study used 4 treatments with 4 replications using a solution of dragon fruit (Hylocereus polyrhizus) and turmeric (Curcuma domestica Val) and 2 positive and negative controls. The positive control used a synthetic dye, namely safranin, and the negative control used no dye. Data analysis in this study was presented in the form of non-statistical qualitative descriptive analysis. Based on the results of the study, the durability of preparations for 0 days showed good tissue staining results, the durability of preparations for 2 days showed results of tissue staining that were still clearly visible, the durability of preparations for 3 days showed results of tissue staining that were no longer clear. The results of this study were declared "feasible" to be used as a medium for learning biology in class XI semester 1 senior high school for exploring cells.
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Lima, Marcus Vinícius Viégas, Abner de Oliveira Freire, Emerson Lucas Frazão Sousa, et al. "Therapeutic Use of Scoparia dulcis Reduces the Progression of Experimental Osteoarthritis." Molecules 24, no. 19 (2019): 3474. http://dx.doi.org/10.3390/molecules24193474.
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Pain is recognized as one of the main symptoms in knee osteoarthritis and is the main reason why patients seek medical attention. Scoparia dulcis has been popularly used to relieve discomfort caused by various painful conditions. The objective of the study is to evaluate the analgesic and anti-inflammatory effect of the crude extract of S. dulcis, in an experimental model of osteoarthritis. The experiment was performed with Wistar rats divided into 4 groups with 5 animals each: healthy, saline, crude extract, and meloxicam groups. Knee osteoarthritis was induced by intra-articular injection of sodium mono-iodoacetate. First, clinical parameters of pain were assessed at days 0, 5, 10, 15, and 20 after induction. Second, the potential cyclooxygenase inhibition was evaluated, and the cytokines of the synovial fluid were quantified. An in silico test and Molecular Docking tests were performed. A histopathological evaluation was made on articular cartilage with safranin O staining. The results showed that a 15-day treatment with crude extract reduced edema, spontaneous pain, peripheral nociceptive activity, and proinflammatory cytokines in the synovial fluid. The highest inhibition of cyclooxygenase 2 in the crude extract occurred at 50 µg/mL. The crude extract of S. dulcis presents therapeutic potential for the treatment of osteoarthritis due to its anti-inflammatory and anti-nociceptive action.
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Lüderitz, Luise, Tilo Dehne, Michael Sittinger, and Jochen Ringe. "Dose-Dependent Effect of Mesenchymal Stromal Cell Recruiting Chemokine CCL25 on Porcine Tissue-Engineered Healthy and Osteoarthritic Cartilage." International Journal of Molecular Sciences 20, no. 1 (2018): 52. http://dx.doi.org/10.3390/ijms20010052.
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Thymus-expressed chemokine (CCL25) is a potent cell attractant for mesenchymal stromal cells, and therefore it is a candidate for in situ cartilage repair approaches focusing on the recruitment of endogenous repair cells. However, the influence of CCL25 on cartilage is unknown. Accordingly, in this study, we investigated the effect of CCL25 on tissue-engineered healthy and osteoarthritic cartilage. Porcine chondrocytes were cultured in a three-dimensional (3D) micromass model that has been proven to mimic key-aspects of human cartilage and osteoarthritic alterations upon stimulation with tumor necrosis factor-α (TNF-α). Micromass cultures were stimulated with CCL25 (0, 0.05, 0.5, 5, 50, 500 nmol/L) alone or in combination with 0.6 nmol/L TNF-α for seven days. Effects were evaluated by life/dead staining, safranin O staining, histomorphometrical analysis of glycosaminoglycans (GAGs), collagen type II (COL2A1) real-time RT-PCR and Porcine Genome Array analysis. 500 nmol/L CCL25 led to a significant reduction of GAGs and COL2A1 expression and induced the expression of matrix metallopeptidases (MMP) 1, MMP3, early growth response protein 1 (EGR1), and superoxide dismutase 2 (SOD2). In concentrations lower than 500 nmol/L, CCL25 seems to be a candidate for in situ cartilage repair therapy approaches.
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Chiu, K. Y., J. Xie, W. K. Ngai, K. S. E. Cheah, T. Kuffner, and S. P. Chow. "NEOCHONDROGENESIS FOR ARTICULAR BONE-CARTILAGE DEFECT: The use of free periosteal graft, porous metal mould and continuous passive motion in a rabbit model." Hand Surgery 01, no. 01 (1996): 79–88. http://dx.doi.org/10.1142/s0218810496000142.
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Full thickness articular bone-cartilage defects were created in the acetabulae of 35 adult rabbits. A double-layered titanium mesh was used in each hip so as to substitute for the bone defect. Free periosteal grafting was then sutured over the mesh, and the hip was subjected to continuous passive motion for 1 week in each rabbit. Under light microscopy, islands of chondroid tissues were shown to be present from 2 weeks onwards, and the dominant reparative tissue was hyaline-like cartilage after 6 months. The normal degree of metachromasia of the matrix by Safranin-0 staining was achieved in most but not all the specimens that were hyaline-like under the microscope. Analysis of the collagen types synthesised by the grafts revealed a combination of both type II and type I collagens. Immunohistochemical staining showed intense positive staining around the chondrocyte lacunae when stained with anti-type II collagen antibodies with the one-year group of rabbits. Although metachromasia of the matrix and collagen typing suggested that fibrocartilage was formed in addition to the hyaline cartilage, the gross appearance and nature of reparative tissues formed were quite promising. Periosteal grafting over a metallic, non-biological surface that provided the shape of the osseous defect in a massive articular defect was therefore a possible alternative.
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Faluti, Arman. "Pemanfaatan Asam Nitrat Sebagai Larutan Pelunak Organ Tumbuhan pada Metode Parafin." Indonesian Journal of Laboratory 5, no. 3 (2022): 98. http://dx.doi.org/10.22146/ijl.v5i3.78678.
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Metode parafin merupakan cara pembuatan preparat permanen dengan menggunakan parafin sebagai media embedding pada organ yang relatif berukuran kecil dan lunak yang diiris dengan ketebalan kurang lebih 6-8 mikron. Sampel yang keras seperti ranting perlu dilakukan proses pelunakan terlebih dahulu agar bagian organ tersebut dapat dipotong dengan mikrotom putar. Asam nitrat sudah digunakan pada beberapa penelitian diantaranya untuk melunakkan tulang pipa tikus. Organ ranting Bougainvillea yang keras direndam dalam asam nitrat selama 24 jam pada berbagai konsentrasi. Tujuan penelitian menguji kemampuan asam nitrat pada berbagai konsentrasi dalam melunakkan organ ranting Bougainvillea serta mengevaluasi irisan dan hasil preparat. Tahapan dalam pembuatan preparat awetan metode parafin adalah: pengambilan organ, fiksasi, perlakuan perendaman asam nitrat selama 24 jam, pencucian (washing), dehidrasi, penjernihan (clearing), infiltrasi, embedding, pengirisan (sectioning), penempelan (affixing) dan pewarnaan (staining). Pita parafin pada perlakuan asam nitrat 0% hampir seluruh bagian preparat pecah, diikuti pada perlakuan 10% dan 15% masing-masing kurang dari setengah sampai seperempat bagian pecah, sedangkan pada perlakuan 20% preparat relatif utuh. Preparat yang sudah diwarnai dengan safranin 1% pada perlakuan perendaman asam nitrat 20% relatif utuh, hanya sebagian kecil (kurang dari 5%) pada bagian kulit kayu dan empulur tidak utuh. Kesimpulan dari penelitian ini penggunaan asam nitrat 20% dapat digunakan untuk melunakkan ranting Bougainvillea pada metode parafin
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Utama, Cahya Setya, Sri Sumarsih, and Marikati Nababan. "Kandungan Mikrobiologis Litter Broiler pada Lama Fermentasi yang Berbeda." Jurnal Agripet 22, no. 1 (2022): 79–87. http://dx.doi.org/10.17969/agripet.v22i1.21501.
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ABSTRACT. Tujuan penelitian adalah mengkaji pengaruh lama fermentasi yang berbeda terhadap bakteri asam laktat, bakteri gram positif/negatif, Salmonella dan Escherichia coli litter broiler. Materi penelitian adalah litter broiler 1 kg, mineral mix, starter mix culture, garam, urea, molases masing-masing 60 gram, NaCl fisiologis 0,85%, alkohol 96%, media MRS, SSA, EMBA, aquades, kristal violet, iodine, dan safranin. Rancangan penelitian menggunakan rancangan acak lengkap (RAL) dengan 4 perlakuan dan 4 ulangan, dengan perlakuan litter broiler lama fermentasi yang berbeda T0 (0 hari), T1 (21 hari), T2 (42 hari) dan T3 (63 hari). Parameter penelitian yaitu total bakteri asam laktat (BAL), bakteri gram positif dan negatif, Salmonella, dan Escherichia coli (E. coli). Analisis data menggunakan uji ANOVA, dan jika terdapat perbedaan dilanjutkan dengan uji DMRT, dengan taraf signifikasi 5%. Hasil penelitian menunjukkan bahwa lama fermentasi yang berbeda memengaruhi total bakteri asam laktat (BAL) litter broiler fermentasi. Semakin lama durasi fermentasi, semakin tinggi total BAL litter broiler. Lama fermentasi yang berbeda tidak memengaruhi skor bakteri gram positif dan negatif litter broiler. Bakteri yang tumbuh pada litter broiler fermentasi berasal dari famili Staphylococcaceae (13,95%), Bacillaceae (32,57%), Streptococcaceae (23,26%), Saccharomycetaceae (6,98%), dan Pseudomonadaceae (23,26%). Bakteri gram positif litter broiler fermentasi berbentuk batang, tidak berspora, soliter, duplococcus, sedangkan bakteri gram negatif berbentuk batang dan soliter. Tidak ditemukan bakteri Salmonella sp. dan E. coli pada litter broiler fermentasi. Lama fermentasi yang berbeda mampu meningkatkan kualitas litter broiler, ditinjau dari total BAL. Litter broiler fermentasi berpotensi dijadikan sebagai alternatif bahan pakan, mengandung 1–3 gram positif dan 0 - 1 gram negatif, serta tidak ditemukan bakteri Salmonella sp. dan E. coli. Perlakuan yang direkomendasikan yaitu litter broiler dengan lama fermentasi 42 hari, dengan jumlah bakteri asam laktat sebanyak 2,4 log CFU/g. (Microbiological content of broiler litter at different times fermentation) ABSTRAK. The aim of the study was to examine the effect of different fermentation times on lactic acid bacteria, gram positive/negative bacteria, Salmonella and Escherichia coli litter broilers. The research material is broiler litter 1 kg, mineral mix, starter mix culture, salt, urea, molasses 60 grams each, 0.85% physiological NaCl, 96% alcohol, MRS media, SSA, EMBA, aquades, crystal violet, iodine , and safranin. The study design used a completely randomized design (CRD) with 4 treatments and 4 replications, with broiler litter treatments with different fermentation times T0 (0 days), T1 (21 days), T2 (42 days) and T3 (63 days). The research parameters were total lactic acid bacteria (LAB), gram positive and negative bacteria, Salmonella, and Escherichia coli (E. coli). Data analysis used the ANOVA test, and if there were differences, it was continued with the DMRT test, with a significance level of 5%. The results showed that different fermentation time affected the total lactic acid bacteria (LAB) in fermented broiler litter. The longer the duration of fermentation, the higher the total LAB of broiler litter. Different fermentation time did not affect the score of gram positive and negative bacteria in broiler litter. The bacteria growing in fermented broiler litter came from the family Staphylococcaceae (13.95%), Bacillaceae (32.57%), Streptococcaceae (23.26%), Saccharomycetaceae (6.98%), and Pseudomonadaceae (23.26%). Gram-positive bacteria fermented broiler litter are rod-shaped, non-sporing, solitary, duplococcus, while gram-negative bacteria are rod-shaped and solitary. No bacteria Salmonella sp and E. coli were found in fermented broiler litter. Different fermentation time can improve broiler litter quality, in terms of total LAB. Fermented broiler litter has the potential to be used as an alternative feed ingredient, containing 1-3 grams positive and 0-1 gram negative, and no Salmonella sp. and E. coli. The recommended treatment is broiler litter with a fermentation time of 42 days, with the number of lactic acid bacteria as much as 2.4 log CFU/g.
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Naka, Yoshifumi, Yusuke Hashimoto, Hiroaki Nakamura, and Kunio Takaoka. "REPAIR OF A MENISCUS DEFECT THROUGH CARTILAGINOUS METAPLASIA OF THE AUTOGENOUS TENDON GRAFT BY INJECTING RECOMBINANT HUMAN BONE MORPHOGENETIC PROTEIN-2." Journal of Musculoskeletal Research 14, no. 01 (2011): 1150001. http://dx.doi.org/10.1142/s0218957711500011.
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Recombinant human bone morphogenetic protein-2 (rhBMP-2) (0, 0.1, 1, or 5 μg) was injected into the autogeneous semitendinosus tendon, and the tendon was transplanted to the region of the medial meniscus defect in a rabbit model to repair the defect. Cartilaginous transformation of the tendon by rhBMP-2 was expected under the less-vascularized intra-articular environment. At four and eight weeks after surgery, the left knee joints were harvested, and the morphological changes of the graft were examined by radiological, histological, and immunohistochemical methods. Cartilaginous tissue within the graft was detected by Safranin O staining and immunostaining of Type-II collagen. At four weeks, fibrocartilagenous tissue, together with small ossicles, was consistently noted in tendon autografts that were injected with 1 or 5 μg of rhBMP-2. At eight weeks, the cartilage located at the basal part of the graft adjacent to the joint capsule was partially replaced with ectopic bone in the 1-μg or 5-μg groups. The ossicles might have been formed by vascular invasion into the rhBMP-2-induced cartilage, but the cartilageous structure remained at the peripheral part of the graft and filled the meniscus defect. The experimental results indicate the potential use of rhBMP-2 in regenerating the cartilaginous meniscus if additional methods to suppress vascular invasion into the rhBMP-2–induced cartilage also inhibit ossification.
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Rodrigues, Renata Dias, Lara Reis Gomes, Rafael Rocha de Souza, and Fernando Cristino Barbosa. "COMPARAÇÃO DA EFICIÊNCIA DAS COLORAÇÕES DE ZIEHL-NEELSEN MODIFICADO E SAFRANINA MODIFICADA NA DETECÇÃO DE OOCISTOS DE Cryptosporidium spp. (EUCOCCIDIORIDA, CRYPTOSPORIDIIDAE) A PARTIR DE AMOSTRAS FECAIS DE BEZERROS DE 0 A 3 MESES." Ciência Animal Brasileira 17, no. 1 (2016): 119–25. http://dx.doi.org/10.1590/1089-6891v17i131267.
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Resumo A criptosporidiose bovina é causada principalmente por quatro espécies distintas: Cryptosporidium parvum, Cryptosporidium bovis, Cryptosporidium ryanae e Cryptosporidium andersoni. A espécie Cryptosporidium parvum (Ordem: Eucoccidiorida, Família: Cryptosporidiidae) é considerada de alto potencial zoonótico, podendo infectar humanos por intermédio da eliminação de oocistos tanto pelos bovinos quanto pelo próprio humano. O objetivo desta pesquisa foi verificar a ocorrência de oocistos de Cryptosporidium spp. em amostras fecais de bezerros (75 machos e 77 fêmeas), tendo sido coletadas 152 amostras de fezes de animais do nascimento até os três meses de idade. O material foi submetido às técnicas de coloração de Ziehl-Neelsen modificado e Safranina modificada, as lâminas foram observadas em toda sua extensão ao microscópio óptico para a verificação da presença de oocistos desta enteroparasitose. Os resultados demonstraram 17,1% (26/152) de positividade no total das amostras examinadas e a análise estatística revelou não haver diferença entre o sexo e as técnicas de coloração utilizadas neste estudo. Conclui-se que a infecção por Cryptosporidium spp. esta presente nas propriedades avaliadas, porém são necessários mais estudos para que o risco de infecção seja mensurado adequadamente e medidas profiláticas implementadas.
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Vivacqua, Thiago Alberto, Rafael Dantas Prinz, Naasson Cavanellas, João Maurício Barretto, Eduardo Branco de Sousa, and Diego Pinheiro Aguiar. "Protocolo para captação, transporte e preservação de tecido osteocondral humano." Revista Brasileira de Ortopedia 55, no. 02 (2020): 163–69. http://dx.doi.org/10.1055/s-0039-3400522.
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Resumo Objetivo Elaborar um protocolo para a captação, transporte e preservação de tecido osteocondral humano para utilização em banco de tecidos (BT). Métodos Foram analisados fragmentos osteocondrais com dimensão de 2 cm3 de 5 doadores cadáveres com idades entre 15 e 45 anos. As amostras foram armazenadas em meio de preservação celular contendo: albumina humana, Iscove's e vancomicina preservados à temperatura de 4°C. A concentração de proteoglicanos no meio extracelular foi quantificada pelo uso de Safranina-O, enquanto a análise estrutural do tecido foi avaliada através de estudo histológico com lâminas coradas em hematoxilina-eosina. As imagens obtidas foram analisadas segundo os escore histológicos de Mankin e o escore proposto pela OsteoArthritis Research Society International. As amostras foram analisadas com 0, 15, 30 e 45 dias de preservação. Resultados Os fragmentos osteocondrais estudados apresentaram diminuição progressiva na concentração de proteoglicanos com o aumento do tempo de preservação. Após 30 dias de preservação, foram identificadas alterações estruturais com descontinuidade da camada superficial da cartilagem. Segundo os resultados obtidos pelo escore de Mankin, houve diferença com significância estatística entre 15 e 30 dias de preservação do tecido. Conclusão O protocolo descrito definiu o transporte de joelho em bloco imerso em Ringer Lactato em temperatura controlada a 10°C até sua chegada ao BT. Após o processamento, a solução de preservação foi composta por meio de cultura celular sem soro Iscove's suplementado com albumina humana a 10% e vancomicina 100 µg/mL. O tecido foi preservado à temperatura de 4°C até o momento do transplante caracterizando a preservação a fresco.
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Zhang, Ying, Zhengzheng Li, Ying He, Yinan Liu, Ge Mi та Jinghong Chen. "T-2 toxin induces articular cartilage damage by increasing the expression of MMP-13 via the TGF-β receptor pathway". Human & Experimental Toxicology 41 (січень 2022): 096032712210755. http://dx.doi.org/10.1177/09603271221075555.
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T-2 toxin pre-disposes individuals to osteoarthritis, Kashin–Beck disease (KBD). The major pathological change associated with KBD is the degradation of the articular cartilage matrix. Herein, we investigated the key molecules that regulate T-2 toxin-mediated cartilage degradation. Potential KBD treatments were also investigated. Sprague Dawley rats were divided into the T-2 toxin group and the control group. The T-2 toxin group received 100 ng/g BW/day, whereas the control group received a similar dose of PBS. The expression of matrix metalloproteinase-13 (MMP-13) and TGF-β receptor I/II (TGF-βRI/II) was analyzed using immunohistochemical staining. C28/I2 chondrocytes were exposed to TGF-βRI/II binding inhibitor (GW788388) for 24 h before incubation in different T-2 toxin concentrations (0, 6, 12, and 24 ng/mL for 72 h). The expression of mRNA for TGF-βRI/II, MMP-13 and proteins for MMP-13, and Smad-2 in chondrocytes were analyzed using RT-PCR and western blot, respectively. Safranin O staining revealed that T-2 toxin treatment modulated the expression of articular cartilage matrix. On the other hand, T-2 toxin treatment sharply increased the expression of MMP-13, TGF-βRI, and TGF-βRII in the rat cartilages. Interestingly, blocking the TGF-βRs-smad 2 signaling pathway using GW788388 abrogated the effect of T-2 toxin on upregulating MMP-13 expression. The expression of MMP-13 in chondrocytes induced with T-2 toxin is regulated via the TGF-βRs signaling pathway. As such, inhibiting the expression of TGF-βRs is a potential KBD treatment.
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Husen, Fajar, Nuniek Ina Ratnaningtyas, Siti Samiyarsih, Juni Safitri Muljowati, and Nur Fitrianto. "Soybean Selection Against Cercospora Leaf Blight Disease Caused By Cercospora kikuchii Based on Anatomical Resistance." Biosaintifika: Journal of Biology & Biology Education 14, no. 1 (2022): 90–102. http://dx.doi.org/10.15294/biosaintifika.v14i1.34865.
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Soybean (Glycine max L. Merr.) is the third food crop commodity after rice and maize in Indonesia. This plant is also known as the most important source of vegetable protein, which is relatively inexpensive, but a decrease in soybean productivity can occur due to infection with disease-causing pathogens, one of is Cercospora kikuchii which causes Cercospora leaf blight (CLB). The research objectives were to determine the anatomical resistance and disease severity of soybean cultivars against CLB. The method was an experiment with a completely randomized design (CRD) factorial pattern; factor 1 being soybean cultivars (Dering, Slamet, Grobogan, Wilis) and factor 2, namely pathogen inoculation (0 conidiospores/mL and 105 conidiospores/mL). Anatomical method preparations using paraffin, staining with 1% safranin. Disease criteria are based on the council of scientific and industrial research (CSIR) assessment method. Data were analysis used analysis of variance (p0.05) and the least significance difference (LSD). The results showed that Dering and Slamet cultivars had the largest cuticle, epidermis, and palisade ratios and the smallest stomata length and width with the largest number of stomata and trichomes compared to Grobogan and Wilis. The disease severity (DS) of the cultivars Dering 14.6%, Slamet 24.64%, Grobogan 24.80% (classified as a resistant with low infection), while Wilis cultivar was 31.08% as a moderately susceptible cultivar with moderate infection. The novelty of soybean cultivar selection against CLB is important and its effectiveness for increasing soybean productivity. Dering, Slamet and Grobogan are likely to be further developed with their resistance to CLB disease.
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Zhao, Zhenzhen, Fengwu Chen, and Yi Wu. "Thiol isomerase ERp72 enhances the development of antibody-induced arthritis." Journal of Immunology 198, no. 1_Supplement (2017): 217.12. http://dx.doi.org/10.4049/jimmunol.198.supp.217.12.
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Abstract Thiol isomerases control labile disulfide bonds that are important in cell activation, cytokine signaling and in a variety of diseases. However, their role in autoimmune diseases such as arthritis remains unknown. ERp72, a thiol isomerases which contains three highly conserved CGHC active motifs, is capable of oxidizing, reducing and isomerizing disulfide bonds. In this study, we generated a new knockout mouse strain deficient of ERp72 (ERp72−/−). Using K/BxN serum-transfer-induced arthritis model (STIA), we determined the role of ERp72 in the pathogenesis of arthritis. To induce STIA, ERp72+/+and ERp72−/− mice received intraperitoneal injection of 200μl K/BxN serum twice on day 0 and day 2. Severity of arthritis was evaluated by daily measurements of joint diameter and clinical scores. Compared with ERp72+/+ mice, ERp72−/− mice developed much more severe arthritis, their joint swelling diameter was significantly increased. Clinical scoring of hind paw edema and hyperemia showed similar results. As indicated by H&E and Safranin O staining, the joint tissue from ERp72−/− mice exhibits more severe synovial hyperplasia, inflammatory cell infiltration and bone erosion on day 12. To determine the role of ERp72 in regulation of cytokine production, we measured the profiles of proinflammatory cytokines. In the joint tissue of ERp72−/− mice bearing STIA, both IL-1β and IL-6 levels were markedly increased by more than 50% than that of ERp72+/+ mice. Interestingly, the basal level of proinflammatory cytokine TNF-α in ERp72−/− mice was significantly higher in that of ERp72−/− mice. Taken together, our observation demonstrates that ERp72 is a negative regulator of innate immunity and plays an important role in the development of arthritis.
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Vinceviča, Amanda Anna, G. R. Devesh Krishnan, and Zanna Martinsone. "THE EFFECT OF HAND DRYERS ON RESTROOM INDOOR AIR QUALITY." ENVIRONMENT. TECHNOLOGY. RESOURCES. Proceedings of the International Scientific and Practical Conference 1 (June 11, 2025): 595–600. https://doi.org/10.17770/etr2025vol1.8678.
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The widespread use of hand-dryers in public restrooms raises concerns about hygiene and indoor air quality. Hand-dryers facilitate microbial proliferation and aerosolization, increasing infection risk and degrading indoor air quality by dispersing pathogens and allergens. This study investigates microbial growth and air contamination, advancing the field by exploring factors such as humidity, airflow mechanics, and particle count concerning various hand-dryer types. Different models of 8 warm air dryers and 2 jet dryers in ten restrooms were sampled (4 female, 4 male and 2 accessible) in an academic institution. The study involved air sampling from the hand-dryer outlet for 30 seconds directly onto agar plates and surrounding air sampling using the “SAS SUPER ISO 180” device. Microbiological samples were then cultivated on different media, manually counted and identified. Fungi were identified by native smears and safranin staining, bacteria using VITEK. Particle count was measured before and during hand dryer use with “TSI P-TRAK”, while other variables such as room temperature, humidity, CO2 levels, hand dryer air flow velocity, temperature were also recorded. The data was processed using IBM SPSS. Statistically significant correlations were found: airflow temperature negatively correlated with fungal dispersal on Sabouraud agar (r=-0.747, p<0.05), CO₂ positively correlated with bacterial dispersal on Trypticase soy agar (TSA) (r=0.661, p<0.05), and humidity showed a significant positive correlation with TSA CFU/min (r=0.636, p<0.05). Microbial contamination was detected in all restrooms. Warm air dryers consistently emitted higher bacterial loads than jet dryers across all tested media. Hand-dryer air ranged from 0 to 1360 CFU/min, while restroom air ranged from 0 to 1424 CFU/m3. Most fungi identified were molds (Mucor spp., Penicillium spp., Aspergillus spp.), with 22 yeast colonies. Analysis identified various opportunistic and pathogenic bacteria, including Acinetobacter baumannii, Escherichia coli, Staphylococcus and other species. Average particles count before use was 7831, while during use 11668. The findings indicate that while hand dryers may contribute to bacterial and fungal dispersal, factors such as airflow temperature, humidity, and CO2 levels also play significant roles in microbial behavior.
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Allen, Nicholas B., Bijan Abar, Richard M. Danilkowicz, Virginia B. Kraus, Steven A. Olson, and Samuel B. Adams. "Intra-Articular Synovial Fluid With Hematoma After Ankle Fracture Promotes Cartilage Damage In Vitro Partially Attenuated by Anti-Inflammatory Agents." Foot & Ankle International 43, no. 3 (2021): 426–38. http://dx.doi.org/10.1177/10711007211046952.
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Background: Intra-articular ankle fracture (IAF) causes posttraumatic osteoarthritis (PTOA), but the exact mechanism is unknown. Proinflammatory mediators have been shown to be present in the synovial fluid fracture hematoma (SFFH) but have not been linked to cartilage damage. The purpose of this study was to determine if the SFFH causes cartilage damage and whether this damage can be attenuated by commercially available therapeutic agents. Methods: Synovial fluid was obtained from 54 IAFs and cultured with cartilage discs from the dome of fresh allograft human tali and randomly assigned to one of the following groups: (A) control—media only, (B) SFFH from days 0 to 2 after fracture, (C) SFFH from days 3 to 9, (D) SFFH from days 10 to 14, (E) group B + interleukin 1 receptor antagonist (IL-1Ra), and (F) group B + doxycycline. The cartilage discs underwent histological evaluation for cell survival and cartilage matrix components. The spent media were analyzed for inflammatory mediators. Results: Cartilage discs cultured with SFFH in groups B, C, and D demonstrated significantly increased production of cytokines, metalloproteinases (MMPs), and extracellular matrix breakdown products. Safranin O staining was significantly decreased in group B. The negative effects on cartilage were partially attenuated with the addition of either IL-1RA or doxycycline. There was no difference in chondrocyte survival among the groups. Conclusion: Exposure of uninjured cartilage to IAF SFFH caused activation of cartilage damage pathways evident through cartilage disc secretion of inflammatory cytokines, MMPs, and cartilage matrix fragments. The addition of IL-1Ra or doxycycline to SFFH culture partially attenuated this response. Clinical Relevance: IAFs create an adverse intra-articular environment consisting of significantly increased levels of inflammatory cytokines and MMPs able to damage cartilage throughout the joint. These data suggest that the acute addition of specific inflammatory inhibitors may decrease the levels of these proinflammatory mediators.
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Bonasia, Davide Edoardo, James A. Martin, Antongiulio Marmotti, et al. "Cocultures of Adult and Juvenile Chondrocytes Compared With Adult and Juvenile Chondral Fragments." American Journal of Sports Medicine 39, no. 11 (2011): 2355–61. http://dx.doi.org/10.1177/0363546511417172.
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Background: The use of allogenic juvenile chondrocytes or autologous chondral fragments has shown promising laboratory results for the repair of chondral lesions. Hypothesis: Juvenile chondrocytes would not affect matrix production when mixed with adult chondrocytes or cartilage fragments. Study Design: Controlled laboratory study. Methods: Cartilage sources consisted of 3 adult and 3 juvenile (human) donors. In part 1, per each donor, juvenile chondrocytes were mixed with adult chondrocytes in 5 different proportions: 100%, 50%, 25%, 12.5%, and 0%. Three-dimensional cultures in low-melt agarose were performed. At 6 weeks, biochemical and histologic analyses were performed. In part 2, isolated adult, isolated juvenile, and mixed 3-dimensional cultures (1:1) were performed with chondral fragments (<1 mm), both with low-melt agarose and a hyaluronic acid scaffold. At 2 and 6 weeks, cultures were evaluated with biochemical and histologic analyses. Results: Part 1: Biochemical and histologic analyses showed that isolated juvenile cultures performed significantly better than mixed and isolated adult cultures. No significant differences were noted between mixed cultures (1:1) and isolated adult cultures. Part 2: Biochemical and histologic results at 6 weeks showed that mixed cartilage fragment cultures performed better than isolated adult cultures in terms of proteoglycans/DNA ratio ( P = .014), percentage of safranin O–positive cells ( P = .012), Bern score ( P = .001), and collagen type II. No statistically significant difference was noted between juvenile and mixed cultures. Conclusion: Extracellular matrix production of juvenile chondrocytes is inhibited by adult chondrocytes. The addition of juvenile cartilage fragments to adult fragments improves matrix production, with a positive interaction between the 2 sources. Clinical Relevance: Even if the underlying mechanisms are still unknown, this study describes the behavior of juvenile/adult cocultures using both chondrocytes and cartilage fragments, with potential for new research and clinical applications.
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Adams, Samuel B., Nicholas B. Allen, and Bijan Abar. "The Intra-Articular Hematoma Immediately after Ankle Fracture Causes Cartilage Damage That is Partially Attenuated by Anti-Inflammatory Agents." Foot & Ankle Orthopaedics 5, no. 2 (2020): 2473011420S0000. http://dx.doi.org/10.1177/2473011420s00002.
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Category: Ankle Arthritis; Basic Sciences/Biologics Introduction/Purpose: Post-traumatic osteoarthritis (PTOA) is a frequent cause of disability. The most common predisposing factor for ankle PTOA is intra-articular ankle fracture. It has been hypothesized that an early inflammatory response after intra- articular injury could lead to cartilage damage. Our group recently demonstrated significantly elevated inflammatory cytokines in the synovial fluid fracture hematoma (SFFH) immediately after intra-articular ankle fracture. The negative effect that this inflammatory environment has on cartilage is unknown but may be the reason for PTOA development. The purpose of this study was to determine if cartilage occurred when exposed to intra-articular SFFH and if anti-inflammatory agents could attenuate this damage. Methods: SFFH was obtained from 52 intra-articular ankle fractures at the time of surgery. Samples were divided into three groups based on time from fracture: acute (0-2 days), intermediate (3-9 days), late (>=10 days). A biopsy punch was used to create 5mm cartilage discs from the medial and lateral shoulders of fresh human talus cartilage. The cartilage discs were weighed and cultured in standard chondrocyte media (SCM) for 3 days. Next, the discs were randomly assigned to one of 4 groups (n=10 each) and cultured as follows for 6 days: A=control, SCM only; Group B=acute SFFH+SCM; Group C=intermediate SFFH+SCM; Group D=late SFFH+SCM. After 6 days of culture, the discs were rinsed and transferred to new wells and cultured for 3 more days in SCM only, to determine inflammatory cytokine and MMP release from the cartilage discs (cartilage damage). The post SFFH spent culture media was collected. The initial and post SFFH culture spent media were analyzed for inflammatory cytokines, MMPs, CTXII (cartilage breakdown marker), and sGAG. Safranin-O staining was then performed on the discs. The initial and post SFFH media from groups A, B, C, and D were compared to determine if there was a difference in damage caused by SFFH based on time from fracture. Based on the results, Group B (early) was felt to cause the most damage. Therefore, the following treatment groups were created and cultured exactly as before: Group E=same as Group B + IL-1Ra treatment; Group F=same as Group B + doxycycline treatment. Groups B, E, and F were compared to determine if the addition of IL-1Ra or doxycycline (known MMP inhibitor) attenuated the damage. Results: Early, intermediate, and late SFFH all caused some degree of cartilage damage in the form of significantly elevated inflammatory cytokine and MMP production from the cartilage discs ( Figure 1 ). The early SFFH (Group B) was noted to cause significantly elevated production of IL-6, IL-8, CTXII, and decreased safranin-O staining. Therefore, it was chosen to compare with the therapeutic anti-inflammatory agents. The addition of IL-1Ra or doxycycline significantly reduced the production of IL-6, IL-8, CTXII, and MMPs 3, 9, and 10 ( Figure 2 ). Conclusion: This is the first study to show that intra-articular hematoma immediately after ankle fracture causes cartilage damage that can be partially attenuated by current clinically available therapeutic agents. Additionally, further research should be performed regarding the safety and efficacy of anti-inflammatory agents to prevent ankle PTOA.
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Bertone, Alicia L., C. Wayne McIlwraith, Robert L. Jones, Robert W. Norrdin, and M. Judith Radin. "Povidone-iodine lavage treatment of experimentally induced equine infectious arthritis." American Journal of Veterinary Research 48, no. 4 (1987): 712–15. https://doi.org/10.2460/ajvr.1987.48.04.712.
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SUMMARY Both tarsocrural joints of 4 horses were inoculated with 1.5 × 105 colony-forming units of Staphylococcus aureus. On days 1, 3, and 6, each horse had one tarsocrural joint lavaged with a balanced electrolyte solution and had the contralateral tarsocrural joint lavaged with 0.1% povidone-iodine solution. All horses were orally administered trimethoprim (5 mg/kg)/sufadiazine (25 mg/kg) combination twice daily and phenylbutazone (2 g) once daily for the duration of the study (21 days). On days 0, 1, 3, 6, 9, 14, and 21, synovial fluid specimens were collected and analyzed for color, clarity, total protein concentration, wbc count and differential, and mucin clot-forming ability. Synovial fluid specimens collected on days 1, 3, 6, 9, 14, and 21 were bacteriologically cultured. On day 21, all horses were euthanatized, the tarsocrural joints were opened and examined, synovial membrane specimens were collected, bacteriologically cultured, and histologically evaluated, and articular cartilage specimens were histochemically evaluated. Repeated measures analysis of variance were used to evaluate differences between lavage solutions and among days for objective measurements. A paired t test was used to evaluate differences between solutions for the indices of synovial membrane inflammation and articular cartilage staining intensity with safranin-O-fast green. To be considered significant, the probability of a type-I error was < 0.05. Significant differences were not found between joints lavaged with electrolyte solution vs povidone-iodine solution for synovial total protein concentration, wbc count, results of synovial fluid and membrane bacteriologie culture, synovial membrane inflammation, or articular cartilage glycosaminoglycan concentration. Generally, more synovial membrane surface fibrin was found in joints lavaged with balanced electrolyte solution than in joints lavaged with povidone-iodine solution. Therefore, use of 0.1% povidone-iodine did not have an advantage over use of balanced electrolyte solution as an irrigation solution in through-and-through joint lavage for the treatment of infecious arthritis in the horse.
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Bertone, Alicia L., D. Michael Davis, Hollis U. Cox, et al. "Arthrotomy versus arthroscopy and partial synovectomy for treatment of experimentally induced infectious arthritis in horses." American Journal of Veterinary Research 53, no. 4 (1992): 585–91. http://dx.doi.org/10.2460/ajvr.1991.53.04.585.
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Summary To evaluate the clinical, laboratory, and histologic effects of 2 methods of treatment for infectious arthritis in horses, Staphylococcus aureus (3.4 to 3.9 × 103 colony-forming units) was inoculated into the tarsocrural joints of 8 horses on day 0. Each horse was treated with phenylbutazone (2 g, po, q 24 h) and gentamicin sulfate (2.2 mg/kg of body weight, iv, q 8 h) for 14 days. On day 2, general anesthesia was induced, and each horse had 1 tarsocrural joint treated by arthrotomy, with removal of accessible fibrin and lavage with 3 L of sterile balanced electrolyte solution. An indwelling plastic drain was placed in the standing horse to provide a means for lavage with 3 L of balanced electrolyte solution twice daily for 72 hours. The contralateral tarsocrural joint was treated via arthroscopic debridement, synovectomy, and lavage with 3 L of balanced electrolyte solution. Arthrotomy and arthroscopic portals were allowed to heal by second intention. Lameness and thermographic examinations, analysis and bacteriologic culture of synovia, cbc, and wbc differential count were performed prior to inoculation and on days 1, 3, 6, 8, and 13. On day 14, each horse was euthanatized, and the joints were measured, opened, and photographed. Synovium and articular cartilage were obtained for semiquantitative histologic (H&E stain) and histochemical (safranin O fast green stain) evaluation. Lameness and joint circumference were significantly (P < 0.05) greater in limbs treated by arthroscopy, synovectomy, and lavage. Arthrotomy with lavage eliminated the S aureus infection significantly (P < 0.05) earlier than arthroscopy, synovectomy, and lavage; however, both treatments eliminated the infection in all but a single joint. Contamination with other organisms (Streptococcus spp and Enterobacter spp) developed significantly (P < 0.05) more often in joints treated by arthrotomy. These results suggested that arthrotomy with lavage was more effective in eliminating joint infection by providing better drainage than arthroscopy, synovectomy, and lavage; however, arthrotomy had a higher risk of ascending bacterial contamination of the joint.
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Talbi, Abderrahmane, Slimane Merouani, Aissa Dehane, Hana Bouchoucha, Ala Abdessemed, and Mohamed S. O. Belahmadi. "Thermo-Catalytic Persulfate Activation in Tubular Microreactors for Advanced Oxidation of Safranin O: Insights into Process Benefits and Limitations." Processes 13, no. 5 (2025): 1494. https://doi.org/10.3390/pr13051494.
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This study examines the use of a1 mm-diameter tubular microreactor submerged in a temperature-controlled water bath to activate potassium persulfate (KPS) via thermal, Fe2+-catalyzed, and combined thermo-catalytic processes for degrading the persistent textile dye Safranin O (SO). The efficiency of these methods was evaluated under varying conditions, including KPS, dye, and Fe2⁺ flow rates, solution pH, reactor length, and water matrix quality (deionized water, tap water, seawater, and secondary effluent from a wastewater treatment plant (SEWWTP)) across bath temperatures of 30–80 °C. Total organic carbon (TOC) analysis validated the results. Maximum dye conversion (up to 89%) occurred at 70 °C, with no improvement beyond this temperature, mainly due to radical-radical recombination. Longer reactors (2–6 m) enhanced conversion, though this effect diminished at higher temperatures due to efficient thermal activation. Increasing dye flow rates reduced removal efficiency, particularly above 50 °C, highlighting kinetic and mass transfer limitations. Persulfate flow rate increases improved conversion, but a plateau emerged at 80 °C. At lower temperatures (30–40 °C), Fe2+ addition significantly boosted SO conversion in deionized water. Between 40 and 50 °C, conversion rose from 30.27% (0 mM Fe2+) to 85.91% (0.2 mM Fe2+) at 50 °C. At higher temperatures (60–80 °C), conversion peaked at 70 °C for lower Fe2+ concentrations (100% for 0.01–0.05 mM Fe2+), but higher Fe2+ levels (0.1–0.2 mM) caused a decline above 60 °C, dropping to 68.44% for 0.2 mM Fe2+ at 80 °C. Deionized, tap, and mineral water showed similar performance, while river water, secondary effluent, and seawater inhibited SO conversion at lower temperatures (30–60 °C). At 70–80 °C, all matrices achieved efficiencies comparable to deionized water for both thermal and thermo-catalytic activation. The thermo-catalytic system achieved >50% TOC reduction, indicating significant organic matter mineralization. The results were comprehensively analyzed in relation to thermal and kinetic factors influencing the performance of continuous-flow reactors.
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Carter, B. G., A. L. Bertone, S. E. Weisbrode, M. Q. Bailey, J. M. Andrews, and J. L. Palmer. "Influence of methylprednisolone acetate on osteochondral healing in exercised tarsocrural joints of horses." American Journal of Veterinary Research 57, no. 6 (1996): 914–22. http://dx.doi.org/10.2460/ajvr.1996.57.06.914.
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Abstract Objective To evaluate joint function and healing of surgically created full-thickness articular cartilage defects in exercised horses after intra-articular administration of methylprednisolone acetate (MPA; 120 mg) and sterile saline solution in the contralateral limb. Design Experimental investigation. Sample Population 12 healthy, sound, radiographically normal horses with induced full-thickness osteochondral lesions on the medial and lateral trochlear ridges of the tali. Procedure Two 8.4-mm-diameter full-thickness articular cartilage lesions were created in each tarsocrural joint (12 horses [24 tarsocrural joints]); 1 was in a weight-bearing (WB) position and the other in a less weight-bearing (LWB) position. Each horse was maintained on a standardized exercise protocol (stall rest, days 0-6; walking, days 7-12; and treadmill, days 13-42) and evaluated throughout the study for changes in joint circumferences, synovial fluid, radiographs, lameness, and scintigraphy. 6 horses were euthanatized on day 42, and 6 on day 180. Gross morphometric assessment was performed, using an image analysis system on a projected color slide of the defect. The type of repair tissue, based on gross appearance, was expressed as a percentage of the total defect for each osteochondral defect. Histochemical assessment was performed, using safranin-O staining for proteoglycans and an image analysis system to express the area of stain uptake. Histomorphometric assessment was performed on H&E-stamed sections, using an image analysis system. The repair tissue filling the defect was categorized as to tissue type and expressed as a percentage of the total defect area. Synovial membrane specimens were assessed semiquantitatively on H&E-stained sections for changes in character. Significance was established at P < 0.05. Results Joint circumference was significantly increased in the saline, compared with the MPA-treated, limbs on days 7, 12, and 42. Synovial fluid WBC counts were significantly increased in the MPA-treated limbs on day 42. Gross osteochondral defects had a greater percentage of mature repair tissue in saline-treated joints (30.8% LWB, 23% WB), compared with MPA-treated joints (0% LWB, 0% WB) at 42 days. Histomorphometric assessment of the repair tissue indicated significant differences with regard to the quality of repair in the saline-treated (34.% fibrous tissue LWB, 19.4% fibrous tissue WB) versus MPA-treated (2.5% fibrous tissue in LWB and WB) joints at 42 days. Microscopically, the percentage of fibrocartilage in the LWB (MPA, 23.7%; saline, 24.8%) was significantly greater than that in the WB (MPA, 14.6%; saline, 15.4%) site at day 180. The MPA-treated limbs had greater villous hyperplasia, edema, and extent of inflammation within the synovial membrane than did saline-treated limbs (days 42 and 180). Conclusion MPA inhibits the development and maturation of repair tissue at 42 days and incites potential long-term (180 days) detrimental synovial membrane inflammation. Furthermore, a single dose of MPA does not cause long-term detrimental effects (180 days) in quality of repair tissue. (Am J Vet Res 1996;57:914–922)
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Pascual-Garrido, Cecilia, Francisco Rodriguez-Fontan, Masahiko Haneda, et al. "Mesenchymal Stem Cells Delivered in a Novel Cartilage Mimetic Hydrogel for the Treatment of Focal Chondral Lesions in an Equine Animal Model." Orthopaedic Journal of Sports Medicine 7, no. 7_suppl5 (2019): 2325967119S0028. http://dx.doi.org/10.1177/2325967119s00288.
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Objectives: A degradable biomaterial has been developed that resembles the native cartilage biochemical properties, in which stem cells can be seeded, differentiate and develop cartilaginous tissue. The purposes of this study were: 1) to determine if mesenchymal stem cells (MSCs) embedded in this cartilage mimetic hydrogel display in vitro chondrogenesis; 2) to demonstrate that the proposed hydrogel can be delivered in situ; and 3) to determine if the hydrogel ± MSCs supports in vivo chondrogenesis. Methods: A photopolymerizable hydrogel consisting of polyethylene glycol, CVPLSLYSGC, chondroitin sulfate (ChS), CRGDS and TGF-β3 was used. Equine bone marrow-derived MSCs were encapsulated in the hydrogel and cultured for 9 weeks. Compressive modulus was evaluated at day 1 and at weeks 3, 6 and 9. Chondrogenic differentiation was investigated via qPCR, Safranin-O staining and immunofluorescence. Three female horses were used. Two 15-mm width x 5-mm depth osteochondral defects were created bilaterally in the medial femoral condyle of each stifle joint. Five groups were established: Hydrogel (n=3), Hydrogel + MSCs (n=3), Microfracture (MFX, n=1), MFX + Hydrogel (n=3), and MFX + Hydrogel + MSCs (n=2). Repair tissue was evaluated at 6 months post intervention with the following cartilage repair scoring systems: macroscopically, International Cartilage Repair Society (ICRS); and histologically, the Modified O’Driscoll scoring (MODS) and ICRS II (Overall assessment 0%, fibrous -100%, hyaline cartilage).The ICRS parameter is scored using a 100-mm VAS, a score of 0 was assigned for properties considered indicative of poor quality and 100 for good quality. Results: In vitro, there was a significant increase in compressive modulus, collagen II and ChS as confirmation of chondrogenesis and hydrogel degradation. (Figure 1) In vivo, the hydrogel was readily photopolimerized in the defect. Cartilage repair was evident in all groups. As shown in Table 1, red indicates best quality score, blue means a poor quality score, but there was no statistical difference. According to the macroscopic ICRS, the hydrogel + MSCs performed better (P= 0.47). However, the MFX + Hydrogel + MSCs tended to perform better per the MODS (P= 0.61); and ICRS-Overall assessment (P= 0.9). Particularly, MFX showed the lowest score for subchondral bone(SCB) abnormalities (0% = abnormal, P= 0.09) but no inflammation was evident (100% = absent, P= 0.53), whereas the Hydrogel had the highest basal integration (100% = complete integration, P= 0.38) but presented moderate inflammation (Figure 2A). MFX showed SCB abnormalities and vascularization (Figure 2 B). Interestingly, a defect treated with MFX + Hydrogel presented more GAGs, less inflammation (vs Hydrogel) and less SCB abnormalities (vs MFX) (Figure 2C). Overall, the group performing better was MFX + Hydrogel + MSCs. Conclusion: This pilot study provides the first evidence of the ability to photopolymerize this novel hydrogel in situ and assess its ability to provide chondrogenic cues for cartilage repair in a large animal model. The presence of all three balanced factors (MFX, Hydrogel, MSCs) had higher scores per MODS summation and ICRS Overall assessment. Strengths of this study include: comparison of standard MFX therapy of osteochondral defects with a novel cartilage mimetic therapy; and use of a large animal that resembles the human knee biomechanically and anatomically. [Figure: see text][Figure: see text][Table: see text]