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Journal articles on the topic 'Immunochemistry – Technique'
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Hurley, Patricia A., Maria Clarke, Jeremy M. Crook, Andrew K. Wise, and Robert K. Shepherd. "Cochlear immunochemistry—a new technique based on gelatin embedding." Journal of Neuroscience Methods 129, no. 1 (October 2003): 81–86. http://dx.doi.org/10.1016/s0165-0270(03)00211-5.
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Sekine, J., A. Irie, K. Sano, K. Hideshima, M. Uehara, and T. Inokuchi. "Application of the membrane filter technique to bromodeoxyuridine immunochemistry for exfoliative cytology." Biotechnic & Histochemistry 76, no. 3 (January 2001): 133–36. http://dx.doi.org/10.1080/bih.76.3.133.136.
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Sekine, J., A. Irie, K. Sano, K. Hideshima, M. Uehara, and T. Inokuchi. "Application of the membrane filter technique to bromodeoxyuridine immunochemistry for exfoliative cytology." Biotechnic and Histochemistry 76, no. 3 (May 1, 2001): 133–36. http://dx.doi.org/10.1080/714028141.
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FITZGERALD, A. M. P., J. J. R. KIRKPATRICK, I. T. H. FOO, and I. L. NAYLOR. "A Picropolychrome Staining Technique Applied to Dupuytren’s Tissue." Journal of Hand Surgery 20, no. 4 (August 1995): 519–24. http://dx.doi.org/10.1016/s0266-7681(05)80167-6.
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Although the histology of Dupuytren’s tissue is well-documented, conventional stains do not distinguish between the different types of collagen which biochemistry and immunochemistry suggest are present. Dupuytren’s specimens [nodules ( n = 26), cords ( n = 15) and dermofasciectomies ( n = 6)] were stained with haematoxylin and eosin, Van Gieson’s, Mallory’s, Masson’s, and Herovici’s picropolychrome stain, and examined for both cellularity and collagen staining characteristics. All stains illustrated the marked cellularity of the nodules, contrasting with a paucity of cells within the cords. The first four stains demonstrated uniformity of the collagen staining within the tissues. Herovici’s picropolychrome, however, showed distinct staining patterns for the dermis, nodules and cords, with both purple/red and blue areas. Other studies suggest that those fibres stained purple/red and blue are types I and III collagens respectively. These findings may shed further light on the tissue of origin of Dupuytren’s disease.
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Lucas, Anne D., Shirley J. Gee, Bruce D. Hammock, and James N. Seiber. "Integration of Immunochemical Methods with Other Analytical Techniques for Pesticide Residue Determination." Journal of AOAC INTERNATIONAL 78, no. 3 (May 1, 1995): 585–91. http://dx.doi.org/10.1093/jaoac/78.3.585.
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Abstract The growing volume of literature concerning immunoassay analysis for trace levels of agrochemicals and other low molecular weight contaminants in various matrixes is indicative of the tremendous interest in and utility of this analytical technique. Most immunoassay methods described in the literature analyze compounds directly, for example, a herbicide in water, or involve solvent exchange of an organic sample extract or dilution of an aqueous-based sample to minimize the matrix effect. As immunoassay for small molecules becomes widely accepted and applied, new challenges involving more complex chemicals in more difficult matrixes arise. The integration of “classical” analytical chemistry with immunochemistry can provide new techniques and approaches useful in discovering the movement, mode of action, and ultimate impact of certain chemicals on humans and the environment.
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Flórez, Luis M. M., Haline F. Ballestero, Anderson P. Duzanski, Paulo R. O. Bersano, João F. Lima, Fernanda L. Cruz, Ligia S. Mota, and Noeme S. Rocha. "Immunocytochemical characterization of primary cell culture in canine transmissible venereal tumor." Pesquisa Veterinária Brasileira 36, no. 9 (September 2016): 844–50. http://dx.doi.org/10.1590/s0100-736x2016000900009.
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Abstract: Immunochemistry with anti-vimentin, anti-lysozyme, anti-alpha 1 antitrypsin, anti-CD3 and anti-CD79α antibodies has been used for characterization of primary cell culture in the transmissible venereal tumor (TVT). Samples for primary cell culture and immunohistochemistry assays were taken from eight dogs with cytological and clinical diagnosis of TVT. To validate the immunochemical results in the primary cell culture of TVT, a chromosome count was performed. For the statistical analysis, the Mann-Whitney test with p<0.05 was used. TVT tissues and culture cells showed intense anti-vimentin immunoreactivity, lightly to moderate immunoreactivity for anti-lysozyme, and mild for anti-alpha-antitrypsin. No marking was achieved for CD3 and CD79α. All culture cells showed chromosomes variable number of 56 to 68. This is the first report on the use of immunocytochemical characterization in cell culture of TVT. Significant statistic difference between immunochemistry in tissue and culture cell was not established, what suggests that the use of this technique may provide greater certainty for the confirmation of tumors in the primary culture. This fact is particularly important because in vitro culture of tumor tissues has been increasingly used to provide quick access to drug efficacy and presents relevant information to identify potential response to anticancer medicine; so it is possible to understand the behavior of the tumor.
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del Agua, Celia, Araceli Rubio-Martinez, Francesc Filipo, Miguel A. Piris, and Pilar Giraldo. "Mycosis Fungoide: Immunochemistry Analysis of Lymphoid and Microenvironment Cells by Macrotissue Array." Blood 110, no. 11 (November 16, 2007): 4400. http://dx.doi.org/10.1182/blood.v110.11.4400.4400.
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Abstract Aims: Cutaneous lymphomas (CL) represent a unique group of lymphomas and are the second most frequent extranodal lymphomas. CL probably is the result of a multifactor and multistep process. Firstly an inflammatory reactive process is developing secondary to various chronic stimuli that may be genetic, environmental, infectious and immunologic. The consequences are the negative effects in cell regulation and deregulation of oncogenes and/or suppressor genes later promotes transition from pre-neoplastic conditions to neoplasia. Detailed molecular expression analysis of cutaneous T-cell lymphoma is not available. Some oncogenic alterations have been demonstrated, such as functional inactivation of the Fas receptor, constitutive activity of STAT 3, or the inactivation of the p16 gene via deletion or promoter hypermethylation. Objective: To study the expression of p16, c-myc, Ki67, bcl-2, CD1a, CD123, TCL1, CD68, STAT 3, STAT 4 and MAL1 in an homogeneous group of. CL diagnosed in a general Hospital in order to know more about the expression of these markers in this neoplastic process. Design: retrospective, analytical study in 30 CL diagnosed consecutively as Mycosis Fungoide (14 early and 16 advanced stages) since January 2005 to December 2006. A macrotissue array technique by incubation with monoclonal antibodies and immunohistochemistry protocol to visualize by avidin-biotin peroxidases was applied in all paraffin histological samples. A semi quantitative method was applied in order to categorize the positive cells. Results: In 28 samples (92%) we have observed p16 positive over expression, the two negative samples corresponding to early affectation. In 14 samples from early stages (48%) c-myc was negative. In 29 samples (96%) CD1a was positive in dermal and epidermal layer. CD123 (interleukine 3 receptor) was negative in 16 samples (52%) and TCL1 was positive in 12 cases (39%) in small cells with oval and cleaved nucleus and scarce cytoplasm. STAT3 and STAT4 were positive in 29 cases (97%), Over expression of MAL1 (48%) was observed in advanced patients with aggressive CL. Conclusions: In our study an over expression of p16 is observed in the majority of cases and high c-myc expression in advanced stages. Probably dendritic plasmacytic cells are involved in the pathogenesis of skin lymphoproliferative disorders with cutaneous T cell infiltration. MAL1 could be a predictor of aggressive CL but it is necessary more studies and more cases in order to confirm it.
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Hu, Xiaotang, Verronika Laguerre, Daniel Packert, Alice Nakasone, and Lynn Moscinski. "A Simple and Efficient Method for Preparing Cell Slides and Staining without Using Cytocentrifuge and Cytoclips." International Journal of Cell Biology 2015 (2015): 1–4. http://dx.doi.org/10.1155/2015/813216.
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Cell staining is a necessary and useful technique for visualizing cell morphology and structure under a microscope. This technique has been used in many areas such as cytology, hematology, oncology, histology, virology, serology, microbiology, cell biology, and immunochemistry. One of the key pieces of equipment for preparing a slide for cell staining is cytology centrifuge (cytocentrifuge) such as cytospin. However, many small labs do not have this expensive equipment and its accessory, cytoclips (also expensive relatively), which makes them difficult to study cell cytology. Here we present an alternative method for preparing a slide and cell staining in the absence of a cytocentrifuge (and cytoclips). This method is based on the principle that a regular cell centrifuge can be used to concentrate cells harvested from cell culture and then deposit the concentrated cell suspension to a slide evenly by using a cell spreader, followed by cell staining. The method presented is simple, rapid, economic, and efficient. This method may also avoid a possible change in cell morphology induced by cytocentrifuge.
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Ivanov, D. P., and A. M. Grabowska. "In Vitro Tissue Microarrays for Quick and Efficient Spheroid Characterization." SLAS DISCOVERY: Advancing the Science of Drug Discovery 23, no. 2 (October 26, 2017): 211–17. http://dx.doi.org/10.1177/2472555217740576.
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Three-dimensional (3D) in vitro microphysiological cultures, such as spheroids and organoids, promise increased patient relevance and therapeutic predictivity compared with reductionist cell monolayers. However, high-throughput characterization techniques for 3D models are currently limited to simplistic live/dead assays. By sectioning and staining in vitro microtissues, researchers can examine their structure; detect DNA, RNA, and protein targets; and visualize them at the level of single cells. The morphological examination and immunochemistry staining for in vitro cultures has historically been done in a laborious manner involving testing one set of cultures at a time. We have developed a technology to rapidly screen spheroid phenotype and protein expression by arranging 66 spheroids in a gel array for paraffin embedding, sectioning, and immunohistochemsitry. The process is quick, mostly automatable, and uses 11 times less reagents than conventional techniques. Here we showcase the capabilities of the technique in an array made up of 11 different cell lines stained in conventional hematoxylin and eosin (H&E) staining, as well as immunohistochemistry staining for estrogen (ER), progesterone (PR), and human epidermal growth factor (Her-2) receptors, and TP53. This new methodology can be used in optimizing stem cell–based models of disease and development, for tissue engineering, safety screening, and efficacy screens in cancer research.
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Wang, Hao, Guo-ming Shen, Wei-jian Liu, Shun Huang, and Meng-ting Zhang. "The Neural Mechanism by Which the Dorsal Vagal Complex Mediates the Regulation of the Gastric Motility by Weishu (RN12) and Zhongwan (BL21) Stimulation." Evidence-Based Complementary and Alternative Medicine 2013 (2013): 1–7. http://dx.doi.org/10.1155/2013/291764.
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A large number of studies have been conducted to explore the mechanism of Back-Shu and Front-Mu points. While several lines of evidence addressed the acupuncture information of Shu acupoints and Mu acupoints gathering in the spinal cord, whether the convergence is extended to the high centre still remains unclear. The study selected gastric Mu points (RN12) and gastric Shu points (BL21) regulating gastric motility and its central neural mechanisms as the breakthrough point, using the technique of immunochemistry, nuclei lesion, electrophysiology, and nerve transection. Here, we report that gastric motility regulation of gastric Shu and Mu acupoints and their synergistic effect and the signals induced by electroacupuncture (EA) stimulation of acupoints RN12 and RN12 gather in the dorsal vagal complex (DVC), increasing the levels of gastrointestinal hormones in the DVC to regulate gastric motility through the vagus. In sum, our data demonstrate an important role of DVC and vagus in the regulation of gastric motility by EA at gastric Shu and Mu points.
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Liu, Sian-Tai, Lin Tsung, Jiun-Lin Horng, and Li-Yih Lin. "Proton-facilitated ammonia excretion by ionocytes of medaka (Oryzias latipes) acclimated to seawater." American Journal of Physiology-Regulatory, Integrative and Comparative Physiology 305, no. 3 (August 1, 2013): R242—R251. http://dx.doi.org/10.1152/ajpregu.00047.2013.
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The proton-facilitated ammonia excretion is critical for a fish's ability to excrete ammonia in freshwater. However, it remains unclear whether that mechanism is also critical for ammonia excretion in seawater (SW). Using a scanning ion-selective electrode technique (SIET) to measure H+ gradients, an acidic boundary layer was detected at the yolk-sac surface of SW-acclimated medaka ( Oryzias latipes) larvae. The H+ gradient detected at the surface of ionocytes was higher than that of keratinocytes in the yolk sac. Treatment with Tricine buffer or EIPA (a NHE inhibitor) reduced the H+ gradient and ammonia excretion of larvae. In situ hybridization and immunochemistry showed that slc9a2 (NHE2) and slc9a3 (NHE3) were expressed in the same SW-type ionocytes. A real-time PCR analysis showed that transfer to SW downregulated branchial mRNA expressions of slc9a3 and Rhesus glycoproteins ( rhcg1, rhcg2, and rhbg) but upregulated that of slc9a2. However, slc9a3, rhcg1, rhcg2, and rhbg expressions were induced by high ammonia in SW. This study suggests that SW-type ionocytes play a role in acid and ammonia excretion and that the Na+/H+ exchanger and Rh glycoproteins are involved in the proton-facilitated ammonia excretion mechanism.
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Johnstone, Alan. "Analytical techniques in immunochemistry." Journal of Immunological Methods 158, no. 2 (February 1993): 281. http://dx.doi.org/10.1016/0022-1759(93)90225-v.
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Shen, Jingling, Zhendong Wang, Xinghui Shen, Zhong Zheng, Qinghua Zhang, Xiuqing Feng, Lili Hu, and Lei Lei. "Abnormal dynamic changes in β-tubulin in somatic nuclear transfer cloned mouse embryos." Zygote 23, no. 1 (December 18, 2013): 76–82. http://dx.doi.org/10.1017/s0967199413000634.
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SummaryThe efficiency of somatic cell nuclear transfer (SCNT) cloning remains low, thus limiting the applications of this technique. In this study, we used immunochemistry and confocal microscopy to detect the microtubule component, β-tubulin, in SCNT, parthenogenetic (PA), and intracytoplasmic sperm injection (ICSI) embryos before the first mitotic division. β-Tubulin is the component subunit of microtubule, which plays critical roles in regulating localization of cellular organelles, and the growth, maturation and fertilization of oocytes. Our results demonstrated similar changes of spindle patterns in PA and ICSI embryos. The second meiotic division resumed 1 h post-treatment, and the cytoplasmic asters (CAs) disappeared. After about 4–6 h of treatment, pronuclei formed with the midbodies connecting each other. Meanwhile, the CAs reappeared and a microtubule network developed in the cytoplasm. However, SCNT embryos showed abnormal multipolar spindles, and the pseudopronuclei that contained many nucleoli existed after 6 h of SrCl2 activation. Enucleated oocytes alone did not form spindle-like structures when they were artificially activated for 6 h, indicating that somatic cell chromosomes might be necessary for spindle formation in SCNT embryos. These results demonstrated abnormal changes of β-tubulin in mouse SCNT embryos, compared with PA and ICSI embryos.
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Leguerney, Ingrid, Ludovic de Rochefort, Marie Poirier-Quinot, Alexandre Ingels, Xavier Violas, Sandra Robin, Paule Opolon, et al. "Molecular Imaging to Predict Response to Targeted Therapies in Renal Cell Carcinoma." Contrast Media & Molecular Imaging 2017 (2017): 1–8. http://dx.doi.org/10.1155/2017/7498538.
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Molecular magnetic resonance imaging targeted to an endothelial integrin involved in neoangiogenesis was compared to DCE-US and immunochemistry to assess the early response of three different therapeutic agents in renal cell carcinoma. Human A498 renal cells carcinoma was subcutaneously inoculated into 24 nude mice. Mice received either phosphate-buffered saline solution, sunitinib, everolimus, or bevacizumab during 4 days. DCE-US and molecular MRI targetingαvβ3 were performed at baseline and 4 days after treatment initiation. PI, AUC, relaxation rate variationsΔR2⁎, and percentage of vessels area quantified on CD31-stained microvessels were compared. Significant decreases were observed for PI and AUC parameters measured by DCE-US for bevacizumab group as early as 4 days, whereas molecularαvβ3-targeted MRI was able to detect significant changes in both bevacizumab and everolimus groups. Percentage of CD31-stained microvessels was significantly correlated with DCE-US parameters, PI (R=0.87,p=0.0003) and AUC (R=0.81,p=0.0013). The percentage of vessel tissue area was significantly reduced (p<0.01) in both sunitinib and bevacizumab groups. We report an early detection of neoangiogenesis modification after induction of targeted therapies, using DCE-US orαvβ3-targeted MRI. We consider these outcomes should encourage clinical trial developments to further evaluate the potential of this molecular MRI technique.
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Diehl, David, Shaffer Mok, Harshit Khara, Amitpal Johal, H. Kirchner, and Fan Lin. "Heparin priming of EUS-FNA needles does not adversely affect tissue cytology or immunohistochemical staining." Endoscopy International Open 06, no. 03 (March 2018): E356—E362. http://dx.doi.org/10.1055/s-0043-121880.
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Abstract Background and study aims Endoscopic ultrasound (EUS)-guided fine-needle aspiration (FNA) or biopsy (FNB) is an indispensable diagnostic tool. Improvements in needling technique have led to increasing tissue yields. Blood clogging of the needle can cause difficulties with specimen handling and stylet passage, which improves when the needle is primed with heparin before use. However, the effect of heparin on cytology, histology or immunochemistry (IHC) of FNA and FNB specimens is unknown. The goal of the study was to evaluate heparin priming on cytologic/histologic appearance, IHC staining, ease of stylet passage, and specimen bloodiness. Patients and methods This was a retrospective study of patients undergoing EUS-FNA/FNB. Needle sizes were 25 gauge (g), 22 g, and 19 g. Heparin priming of the needle was done and the stylet replaced (“dry heparin”) or suction attached without replacing the stylet (“wet heparin”). Smears and cellblocks were examined by pathologists, and IHC staining were done as needed. Specimen bloodiness was compared with matched controls. Results Adequate tissue yields were obtained in all samples (37 heparin, 36 no heparin). Heparin priming did not exhibit negative effects on cytologic or histologic interpretation of the specimens, nor IHC. There was no difference in cellblock bloodiness between the heparin primed needle specimens and the non-heparin control group. Conclusions Heparin priming of EUS-FNA or FNB needles does not negatively affect cytologic or histologic interpretation, nor interfere with IHC. In addition, heparin priming does not increase specimen bloodiness. When the “wet suction” technique is used for EUS-FNA, heparin priming can be used instead of saline priming of the EUS needle.
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Hae Cho, Eun, Sang-Mi Lee, Hyeon-Seok Eom, In-Suk Kim, Gyeong-Won Lee, and Sun-Young Kong. "Fluorescent in Situ Hybridization Analysis at Bone Marrow Biopsy Section in Multiple Myeloma." Blood 114, no. 22 (November 20, 2009): 4898. http://dx.doi.org/10.1182/blood.v114.22.4898.4898.
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Abstract Abstract 4898 Introduction The technique of fluorescence immunophenotyping and interphase cytogenetics as a tool for the investigation of neoplasms has recently been introduced to detect molecular cytogenetic abnormalities in plasma cell myeloma of bone marrow (BM) aspirate. However, in case of sub-optimal BM aspirate or the focal distribution of myeloma in the BM, the plasma cells are significantly lower in the BM aspirate than those of biopsy section. Therefore, we have developed a sensitive fluorescence in situ hybridization (FISH) technique which is combined with immunochemistry and is applicable to BM biopsy section for molecular cytogenetic study of plasma cell neoplasms. Patients and Methods Conventional cytogenetic analysis and FISH results of BM samples of 35 multiple myeloma (MM) patients at the time of diagnosis have been evaluated. The probe for IgH rearrangement has been used for hybridization with myeloma cells coupled with CD138 immunostain at BM biopsy section. Results Nineteen patients (54.3%) had abnormal FISH IgH results in biopsy section, whereas seven (20%) cases had abnormal findings in BM aspirate. FISH IgH analysis at biopsy section revealed various signal patterns and proportions (range 6-87%) of cells with atypical signals out of CD138 positive cells. Among five cases with <10% of plasma cells at BM aspirate, four (80%) had abnormal FISH results at biopsy section, whereas one (20%) had abnormal signals at aspirate. There is no correlation between the proportions of cells with atypical signal corrected by the plasma cell count at BM aspirate and the proportions of cells with atypical signal at biopsy section. Conclusions FISH analysis combined with immunostain which is applied at biopsy section is a highly sensitive and convenient technique to detect and quantify monoclonal plasma cells. It could be used for molecular cytogenetic study in plasma cell neoplasms even though there are less than 10% of plasma cells at BM aspirate and the monitoring of residual disease. Disclosures Kong: National cancer center, Korea: Research Funding.
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Song, Wei, Guiyun Song, Can Zhao, Xiaoguang Li, Xiaojiao Pei, Wen Zhao, Yudan Gao, Jia-Sheng Rao, Hongmei Duan, and Zhaoyang Yang. "Testing Pathological Variation of White Matter Tract in Adult Rats after Severe Spinal Cord Injury with MRI." BioMed Research International 2018 (November 11, 2018): 1–13. http://dx.doi.org/10.1155/2018/4068156.
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The purpose of this study was to assess the pathological variation in white matter tracts in the adult severe thoracic contusion spinal cord injury (SCI) rat models combined with in vivo magnetic resonance imaging (MRI), as well as the effect of spared white matter (WM) quantity on hindlimb motor function recovery. 7.0T MRI was conducted for all experimental animals before SCI and 1, 3, 7, and 14 days after SCI. The variation in the white matter tract in different regions of the spinal cord after SCI was examined by luxol fast blue (LFB) staining, NF200 immunochemistry, and diffusion tensor imaging (DTI) parameters, including fraction anisotropy, mean diffusivity, axial diffusion, and radial diffusivity. Meanwhile, Basso-Beattie-Bresnahan (BBB) open-field scoring was performed to evaluate the behavior of the paraplegic hind limbs. The quantitative analysis showed that spared white matter measures assessed by LFB and MRI had a close correlation (R2 = 0.8508). The percentage of spared white matter area was closely correlated with BBB score (R2 = 0.8460). After SCI, spared white matter in the spinal cord, especially the ventral column WM, played a critical role in motor function restoration. The results suggest that the first three days provides a key time window for SCI protection and treatment; spared white matter, especially in the ventral column, plays a key role in motor function recovery in rats. Additionally, DTI may be an important noninvasive technique to diagnose acute SCI degree as well as a tool to evaluate functional prognosis. During the transition from nerve protection toward clinical treatment after SCI, in vivo DTI may serve as an emerging noninvasive technique to diagnose acute SCI degree and predict the degree of spontaneous functional recovery after SCI.
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Mücke, Marcus M., Dominik Bettenworth, Christiane Geyer, Katrin Schwegmann, Christopher Poremba, Michael Schäfers, Dirk Domagk, Carsten Höltke, and Philipp Lenz. "Targeting Mucosal Endothelin-A-Receptor Expression by Fluorescence Endoscopy is Feasible to Detect and Characterize Colitis-Associated Cancer in Mice." Inflammatory Bowel Diseases 24, no. 1 (December 19, 2017): 111–22. http://dx.doi.org/10.1093/ibd/izx032.
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Abstract Background To facilitate onsite decision-making during endoscopy, both accurate detection and in vivo characterization of preneoplasia are prerequisites. However, no endoscopy technique is available that meets both demands satisfactorily. We evaluated endothelin-receptor A (ETAR)-guided fluorescence endoscopy (FE) in vivo and fluorescence reflectance imaging (FRI) ex vivo for detection and characterization of early dysplastic colitis-associated colonic lesions. Methods Colorectal cancerogenesis was investigated in the inflammatory driven AOM-DSS model and spontaneous adenoma development in ApcMin mice. A Cy5.5-labeled nonpeptidic ETAR-specific imaging probe was injected intravenously to assess tumor development in vivo by white light endoscopy (WLE) and FE. Ex vivo tumors were evaluated by FRI, histological examination, and western blot analysis. In addition, tissue samples from patients with colitis-associated malignant and nonmalignant mucosal alterations were analyzed. Specificity experiments were performed using an unspecific Cy3.5-glycine tracer. Results Overall, 62 adenomas were observed. FE was able to detect and quantify ETAR expression targeting the ETAR-specific photoprobe. A significantly higher fluorescent contrast was detected in colonic adenomas compared to adjacent nonmalignant mucosa by FE (64.3 ± 7.9 vs. 56.6. ± 7.0; P < 0.001). These results were confirmed by FRI examination, immunochemistry, and western blot analysis. Additionally, ETAR expression in samples from human patients with colitis-associated cancer was highly elevated compared to nonmalignant alterations. Specificity experiments indicated a high binding-specificity of the applied ETAR photoprobe (1.4 ± 0.3 vs. 2.5 ± 0.7; P < 0.001). Conclusions We introduced ETAR guided FE in mice for successful in vivo detection and characterization of colorectal neoplasia on a molecular level.
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Tropea, Margaret, Bonnie Harper, Grace Graninger, Terry Phillips, Gabriela Ferreyra, Howard Mostowski, Robert Danner, Anthony Suffredini, and Michael Solomon. "Isolation of a circulating CD45-, CD34dim cell population and validation of their endothelial phenotype." Thrombosis and Haemostasis 112, no. 10 (2014): 770–80. http://dx.doi.org/10.1160/th14-01-0043.
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SummaryAccurately detecting circulating endothelial cells (CECs) is important since their enumeration has been proposed as a biomarker to measure injury to the vascular endothelium. However, there is no single methodology for determining CECs in blood, making comparison across studies difficult. Many methods for detecting CECs rely on characteristic cell surface markers and cell viability indicators, but lack secondary validation. Here, a CEC population in healthy adult human subjects was identified by flow cytometry as CD45-, CD34dim that is comparable to a previously described CD45-, CD31bright population. In addition, nuclear staining with 7-aminoactinomycin D (7-AAD) was employed as a standard technique to exclude dead cells. Unexpectedly, the CD45-, CD34dim, 7-AAD- CECs lacked surface detectable CD146, a commonly used marker of CECs. Furthermore, light microscopy revealed this cell population to be composed primarily of large cells without a clearly defined nucleus. Nevertheless, immunostains still demonstrated the presence of the lectin Ulex europaeus and von Willebrand factor. Ultramicro analytical immunochemistry assays for the endothelial cell proteins CD31, CD34, CD62E, CD105, CD141, CD144 and vWF indicated these cells possess an endothelial phenotype. However, only a small amount of RNA, which was mostly degraded, could be isolated from these cells. Thus the majority of CECs in healthy individuals as defined by CD45-, CD34dim, and 7-AAD- have shed their CD146 surface marker and are senescent cells without an identifiable nucleus and lacking RNA of sufficient quantity and quality for transcriptomal analysis. This study highlights the importance of secondary validation of CEC identification.
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Properzi, Francesca, Emmanuel Comoy, Mariantonia Logozzi, Stefano Fais, Ilaria Cristofaro, Luana Lugini, Massimo Venditti, et al. "Detection of exosomal prions in blood by immunochemistry techniques." Journal of General Virology 96, no. 7 (July 1, 2015): 1969–74. http://dx.doi.org/10.1099/vir.0.000117.
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Giotta, F., G. Simone, V. Fazio, S. Longo, S. Petroni, V. Rubini, M. Liuzzi, T. Addati, and G. Colucci. "Patterns of HER2/neu, hormonal receptor expression, and proliferative activity in primary and metastatic breast cancer." Journal of Clinical Oncology 27, no. 15_suppl (May 20, 2009): 1071. http://dx.doi.org/10.1200/jco.2009.27.15_suppl.1071.
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1071 Background: The modification of biological features of metastatic sites (MS) in breast cancer patients arise some debatable questions regarding clinically usefull information and safe/efficient methods to detect them. Fine needle aspiration (FNA) of visceral MS is an available tool to characterize tumoral lesion and liquid based citology technique provides usefull cell samples for immunocytochemical and/or molecular assays. Methods: The aim of this study was to compare prognostic and predictive factors obtained from primary tumors (PT) and corresponding MS. Fluorescent in situ hybridization (FISH) was performed for HER2/Neu determination, while ER, PgR, and MIB-1 were detected by immunochemistry using specific monoclonal antibodies on monolayered cell sample and in the corresponding cytoinclusion. FNA with a 21–23 G needle was performed in 20 consecutive breast cancer patients with distant metacronous MS. Results: In 8/20 patients, both ER and PgR were absent in PT and in MS, in 7 were both present, in 4 cases only ER was detected, and only 1 case was ER negative with a low PgR. About the proliferative activity (MIB-1 index: cut off value >20%) only 3 MS presented an higher value. With regard to HER2/Neu, 4/20 cases were amplified and no discrepancies were found between cytological and cytoinclusion specimens. No substantial changes were found about kinetic activity. HER2/Neu status as assessed in PT was confirmed in MS in 10/12 cases; a lung mestastasis showed amplification while primary was not amplified and a liver lesion lost the amplification which was detected in the PT. Conclusions: Our study strongly suggest the opportunity in using FNA for detection of prognostic and predictive factors in MS. Pectasides et al. (Anticancer Res. 2006) found in patients with altered or conserved HER2/Neu in PT and in MS different response rates between the two groups and a significant poorer prognosis in patients with altered Neu. We think that liquid-based cytology applied on FNA of distant metastases could help us to understand some more in this issue. No significant financial relationships to disclose.
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Olejnik, J. A., M. Herrid, R. Davey, K. Hutton, G. Hinch, and J. Hill. "263. The successful use of busulfan to deplete endogenous spermatogonia in ram testes." Reproduction, Fertility and Development 17, no. 9 (2005): 107. http://dx.doi.org/10.1071/srb05abs263.
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Our research into germ cell transfer builds upon murine research1 and is aimed at using this technique in livestock species. To increase the efficiency of colonization of transplanted germ cells, the recipient testes must be depleted of endogenous spermatogonia, without affecting the supporting cells. Three depletion methods were investigated, heat, cold and chemotherapy. Our first investigation looked at the direct cooling (0ºC) and heating (45ºC) of the testes of 4–6 week old ram lambs. Testes were collected 7 days post treatment. The second investigation involved the systemic injection of busulfan to ram lambs aged 3-4 months. Busulfan is used for preparing recipient mice for testes cell transfer.1 At doses affecting the stem cells of the testes, busulfan will result in mylosuppression. Therefore a preliminary dose response trial was conducted at dose rates 4, 8 and 16mg/kg to determine the most effective dose, without threatening the survival of the animal. Testes were recovered after 3 and 6 weeks. All testes sections followed routine histology and immunochemistry with PGP 9.5 (Table 1). For the heat and cold study, only gonocytes were present and there were no differences in testes weights, tubule diameters or gonocyte numbers in any of the treatment groups. For the busulfan study, dose rates of 8 and 16 mg/kg resulted in severe mylosuppression and euthanasia of 7 out of 8 animals between day 12 and 18, whereas animals in the 4mg/kg group showed only mild clinical effects, that were not life threatening. These results indicate that busulfan reduced endogenous spermatogonia in the pre-pubertal ram. This effect is observed at systemic doses of 4 mg/kg or higher; however, doses of 8 mg/kg and above are lethal to the survival of the animal. The use of direct heat (45ºC) or cold (0ºC) to the testes does not affect gonocyte numbers in ram lambs; however, effects on more mature stages was not studied. (1)Brinster RL, Zimmermann JW. (1994) Spermatogenesis following male germ-cell transplantation. Proc Natl Acad Sci USA 91, 11298–11302.
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Parkinson-Lawrence, Emma, Maria Fuller, John J. Hopwood, Peter J. Meikle, and Doug A. Brooks. "Immunochemistry of Lysosomal Storage Disorders." Clinical Chemistry 52, no. 9 (September 1, 2006): 1660–68. http://dx.doi.org/10.1373/clinchem.2005.064915.
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Abstract Background: Lysosomal storage disorders are a group of genetic diseases, each with a broad spectrum of clinical presentation that ranges from attenuated to severe. The immunochemical analysis of patient samples is aimed at several key aspects of patient management, including early detection of the disorder, prediction of clinical severity, determining the most appropriate therapeutic regimen, and monitoring of patients on therapy. Methods: In this study, we review the current and emerging technology available to achieve these assessments. Results: Immune assays have direct practical application for the early detection, diagnosis and prognosis of lysosomal storage disorder patients. Multiplexing of these assays may provide a platform to allow newborn screening for multiple lysosomal storage disorders. Conclusions: We have reviewed the immunochemical techniques available for the analysis of lysosomal storage disorder patient samples and advise that these may be used in conjunction with other technologies for effective patient management.
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Bonini, P. A., G. Banfi, M. Pazzagli, G. Messeri, and A. Roda. "Detection, signal processing, and calibration In lmmunoassay systems." Journal of Automatic Chemistry 13, no. 2 (1991): 45–47. http://dx.doi.org/10.1155/s146392469100010x.
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The new trends in immunochemistry related to the replacement of radioisotopic labels with non-radioactive labels are presented. Immunoenzymatic, fluorescent and chemiluminescent techniques are described in terms of their basic principles and their most common applications. The advantages of computer-controlled calibration are also discussed.
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Li, Ke, Yunhua Mao, Yiyuan Li, Ruji Wu, Dejuan Wang, and Jianguang Qiu. "The role of programmed death-ligand 1 and non-epithelial circulating tumor cells in locally advanced prostate cancer." Journal of Clinical Oncology 38, no. 15_suppl (May 20, 2020): e17557-e17557. http://dx.doi.org/10.1200/jco.2020.38.15_suppl.e17557.
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e17557 Background: Circulating tumor cells (CTCs) is considered as an effective prognostic biomarker in malignancies. It is recently recognized that detections of programmed death-ligand 1 (PD-L1) and mesenchymal markers in CTCs may improve the predictive value. Therefore, we aimed to determine the cut-off values of these CTCs biomarkers in prostate cancer (PCa), assess the rates of PD-L1 positive and non-epithelial (NE) subgroup in CTCs among patients with high-risk (HRPC) and locally advanced PCa (LAPC) before surgery, and evaluate the clinical values of these markers. Methods: Four PCa cell lines selected from Cancer Cell Line Encyclopedia were applied in spiking experiments to establish the cut-offs of PD-L1 positive and NE CTCs. 182 HRPC and 89 LAPC patients were enrolled from June 2016 to December 2018. All patients underwent radical prostatectomy after blood sample collection. CTCs enumeration and biomarkers identification were conducted by Canpatrol CTC enrichment system and the RNA in situ hybridization technique. Lastly, using the cut-offs,we evaluated the association between CTCs markers and other variables in terms of clinical data, disease outcome, tumor tissues immunochemistry (IHC) staining, and the response of immunotherapy. Results: The cut-offs were determined as the lowest rates of NE and PD-L1+ CTCs among the cell lines, 45% and 25%, respectively. In HRPC patients, a higher PSA level and more advanced T stage were observed in the NE CTCs positive group (≥45%), compared with the negative group ( P= 0.041 and P= 0.004). Multivariate analysis showed that the NE CTCs rate, instead of total CTCs number, was an independent risk predictor for progression-free survival (PFS) (HR = 3.85, P= 0.013). In LAPC patients, the PD-L1+ CTCs positive group (≥25%) presented a higher PSA level and higher percentage of lymph node metastasis than negative group ( P= 0.027 and P= 0.028). The median PFSs between the two groups were 16.6 mouths vs. 21.8 mouths, respectively. (HR = 2.87, P= 0.002 ). No significant correlation was found between PD-L1 expression in CTCs and in tumor tissue IHC staining. However, we observed a significant correlation between PD-L1+ CTCs rate and ERG or PTEN expression in tissues ( P= 0.010 and P= 0.009). In addition, patients in PD-L1+ CTCs positive group presented a better PSA response to Durvalumab treatment than those in negative group. Conclusions: PD-L1+ and NE CTCs were an effective prognosis predictors for HRPC and LAPC patients with radical prostatectomy. LAPC patients with PD-L1+ CTCs might benefit from adjuvant immunotherapy
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Dunbar, BS, S. Avery, V. Lee, S. Prasad, D. Schwahn, E. Schwoebel, S. Skinner, and B. Wilkins. "The mammalian zona pellucida: its biochemistry, immunochemistry, molecular biology, and developmental expression." Reproduction, Fertility and Development 6, no. 3 (1994): 331. http://dx.doi.org/10.1071/rd9940331.
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Many studies of the molecular and biochemical aspects of mammalian fertilization have focused on the interaction of the spermatozoa with the zona pellucida (ZP). The zona pellucida, a unique extracellular matrix surrounding the mammalian oocyte, is formed during ovarian follicular development. Following ovulation of the mature ovum, the spermatozoa must bind to and penetrate this matrix before the fertilization process is completed and the male and female genetic information combine. Although numerous models for this interaction have been proposed, the complete process has yet to be elucidated. The precise mechanisms by which these interactions occur also vary markedly among different mammalian species, making it more difficult to establish a unified model. To a great extent, the study of the molecules involved in these interactions have been limited because small numbers of female gametes are available for these studies. The recent development of techniques to isolate large numbers of zonae pellucidae as well as advances in immunological and molecular biology techniques have permitted the detailed characterization of ZP proteins. Although there is a paucity of information on the post-translational modification and extracellular processing of these molecules which result in matrix formation, a number of properties have been elucidated allowing better correlation between the structure and function of different ZP proteins among species. This review reflects these studies in relation to protein nomenclature and the molecular complexity of ZP antigens.
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Li, Xuefeng, Tao Li, Li Yang, and Long-Yun Li. "Protective roles of ketamine and xylazine against lightinduced retinal degeneration in rats." Tropical Journal of Pharmaceutical Research 18, no. 4 (May 18, 2021): 727–33. http://dx.doi.org/10.4314/tjpr.v18i4.7.
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Purpose: To study the protective effects of ketamine and xylazine against light exposure-induced retinal degeneration (RD) in rats.
Methods: Sprague Dawley rats were divided into three groups viz: light damage before anesthesia (LAE), light damage only (LDO), and control (CON) group which was kept in the dark for 12 - 18 h to habituate before light exposure. LDO group was exposed to light before anesthesia, while LAE group was maintained under anesthesia with ketamine and xylazine. The groups were kept for 120 min in darkness after anesthesia prior to light exposure and they were awakened prior to light damage. Functional assessment was carried out using electroretinography while morphological analysis was carried out using histology and immunochemistry techniques.
Results: Ketamine-xylazine combination preserved the function of the retina and protected against light-induced RD based on retinal imaging studies and immunochemistry analysis. Xylazine and ketamine anesthesia provided protection against light-induced retinal damage, and thus reduced photoreceptor cell death.
Conclusion: These results indicate that xylazine and ketamine anesthesia offer protection against lightinduced damage and photoreceptor cell death in rats, and therefore, can potentially be developed for use in humans.
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Narkhede, Mayur, Sadaf Qureshi, Maryam Yazdy, Roxanna Juarez, and Giuseppe Esposito. "Lack of Prognostic Significance of Pre-Treatment Total Metabolic Tumor Volume (TMTV) in Diffuse Large B-Cell Lymphoma (DLBCL)." Blood 132, Supplement 1 (November 29, 2018): 1720. http://dx.doi.org/10.1182/blood-2018-99-113791.
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Abstract Background DLBCL is the most common non-Hodgkin lymphoma (NHL), making up about 30%-40% of NHL in the U.S. PET-CT is recommended as the most accurate imaging technique in DLBCL for staging and response assessment. Pretreatment assessment of PET-CT scan derived metrics such as TMTV has been shown to correlate with PFS and/or overall survival (OS) in DLBCL (Sasanelli 2014) We attempted to replicate this finding using EFS at 24 months as a primary endpoint and compare it with pre-treatment TMTV, TLG and cell of origin (COO). Methods 47 pts with newly diagnosed DLBCL and treated with R-CHOP at our institution between 2014 to 2018 were identified from our electronic medical record system for retrospective analysis after IRB approval. All pts had a pretreatment PET-CT scan available for TMTV measurement. All pts had a pretreatment biopsy which were reviewed along with their clinical information regarding treatment outcome and follow up. Patients were classified as to germinal center B cell (GCB) and non-GCB based on immunochemistry using the Hahn's algorithm. PET-CT scans were reviewed by two nuclear medicine physicians using synovia software, and measurements for TMTV and TLG were recorded. TMTV was calculated using a threshold of 41% of the max pixel value (based on prior studies) to draw the volume of interest (VOI) for a lesion. Pooled t-test was performed to compare TMTV, TLG and COO with EFS at 24 mos. Chi-Square test compared TMTV with COO Results Median age of pts was 58 years, with a median duration of follow up of 26 months. There were 33% with limited stage (Stage I or II) and 67% were advanced stage (Stage III or IV). The mean pretreatment TMTV and pretreatment TLG was 295cm3 and 4519 units. 49% were GCB subtype and 47 % non-GCB. Amongst all patients 19.2 % had an event within 24 mos. When TMTV was compared to EFS at 24 months the mean TMTV was 304 for those who had an event versus 294 without (p=0.95). TLG compared to EFS at 24 months showed a mean TLG of 3391 for those who had an event versus 4914 without (P=0.40). GCB and non-GCB had mean TMTV of 264 and 339 respectively with p =0.59. COO when compared to TLG had means of 4365 and 4933 for GCB and non-GBB respectively with p=0.79.Whereas there was no correlation between stage and COO (p=0.4296) TMTV correlated with Ann Arbor staging (p=0.0002). Conclusion This retrospective study failed to demonstrate a correlation between pre-treatment TMTV, TLG, COO and EFS at 24 months revealing the lack of prognostic significance of pretreatment PET scan derived metrics in DLBCL. Prior studies with TMTV did not evaluate EFS at 24 months as an endpoint and therefore, longer follow up might be needed to demonstrate prognostic significance of pretreatment TMTV minimizing it clinical significance. The different subtypes of DLBCL based on COO as assessed by Hahns algorithm also did not differ in their disease burden as measured by TMTV. Disclosures No relevant conflicts of interest to declare.
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Clemetson, K. J., J. L. McGregor, R. P. McEver, Y. V. Jacques, D. F. Bainton, W. Domzig, and M. Baggiolini. "Absence of platelet membrane glycoproteins IIb/IIIa from monocytes." Journal of Experimental Medicine 161, no. 5 (May 1, 1985): 972–83. http://dx.doi.org/10.1084/jem.161.5.972.
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Two-dimensional gel electrophoresis, immunoprecipitation, and crossed immunoelectrophoresis were used in the investigation of glycoproteins IIb/IIIa in platelets, monocytes, and monocyte-derived macrophages from human blood. All techniques detected the glycoproteins in platelets but not in the mononuclear phagocytes. Similar results were obtained by immunochemistry using a monoclonal antibody against the platelet glycoproteins IIb/IIIa (revealed by a gold-labeled second antibody) which bound heavily to the platelet but not to the monocyte surface. The biochemical techniques used for the analysis of mononuclear phagocytes would have reliably detected the level of glycoproteins IIb/IIIa contributed by a 5% contamination with platelets, calculated on a per cell basis. We conclude that human monocytes and monocyte-derived macrophages lack glycoproteins IIb/IIIa. Our results further indicate that centrifugal elutriation yields monocyte preparations with minimal contamination by platelets. It seems likely that the positive results obtained by other authors were due to the presence of platelets or fragments on the monocytes.
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Fayon, Michael, Eric Dumas De La Roque, Patrick Berger, Hugues Begueret, Olga Ousova, Mathieu Molimard, and Roger Marthan. "Increased relaxation of immature airways to β2-adrenoceptor agonists is related to attenuated expression of postjunctional smooth muscle muscarinic M2 receptors." Journal of Applied Physiology 98, no. 4 (April 2005): 1526–33. http://dx.doi.org/10.1152/japplphysiol.00948.2004.
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Spontaneous or agonist-induced contraction of airway smooth muscle can be observed very early in fetal life, thus explaining the possible occurrence of bronchospasm in very low birth weight infants within the first days of life. In an attempt to better manage such bronchospasms, the aim of the present study was to investigate the age-specific modifications in airway smooth muscle relaxation to β2-agonists and muscarinic antagonists using a combination of functional and molecular techniques. In the rat, isometric relaxation to the β2-agonist salbutamol was examined in tracheae; we also examined muscarinic receptor expression (M2R and M3R mRNA levels) in airway smooth muscle by immunochemistry, Western blotting, and real-time PCR. Compared with adults, salbutamol-induced relaxation was twofold greater in immature rat isolated tracheae preconstricted by carbachol. This effect was associated with a lower expression of M2R in the smooth muscle of immature animals (sixfold and almost twofold as assessed by immunochemistry and Western blotting, respectively). Real-time PCR data indicate that changes in M2R expression according to age occurred at a posttranscriptional level. In adult airways, there was a significantly greater functional efficacy of M2R blockade by methoctramine compared with that shown in immature rats. Because of the limited availability of human neonate lung tissue, only the molecular part of the study was performed, and we observed a qualitatively similar effect, i.e., a lower M2R expression in the neonatal airway smooth muscle, although this was quantitatively smaller. We conclude that β2-agonist-induced relaxation is enhanced in immature compared with adult airways as a result of greater postjunctional M2R expression in adult airway smooth muscle. This finding may be of importance in the clinical management of bronchoconstriction in neonates.
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Klein, Reinhild, Douglas M. Templeton, and Michael Schwenk. "Applications of immunochemistry in human health: advances in vaccinology and antibody design (IUPAC Technical Report)." Pure and Applied Chemistry 86, no. 10 (October 21, 2014): 1573–617. http://dx.doi.org/10.1515/pac-2013-1028.
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Abstract This report discusses the history and mechanisms of vaccination of humans as well as the engineering of therapeutic antibodies. Deeper understanding of the molecular interactions involved in both acquired and innate immunity is allowing sophistication in design of modified and even synthetic vaccines. Recombinant DNA technologies are facilitating development of DNA-based vaccines, for example, with the recognition that unmethylated CpG sequences in plasmid DNA will target Toll-like receptors on antigen-presenting cells. Formulations of DNA vaccines with increased immunogenicity include engineering into plasmids with “genetic adjuvant” capability, incorporation into polymeric or magnetic nanoparticles, and formulation with cationic polymers and other polymeric and non-polymeric coatings. Newer methods of delivery, such as particle bombardment, DNA tattooing, electroporation, and magnetic delivery, are also improving the effectiveness of DNA vaccines. RNA-based vaccines and reverse vaccinology based on gene sequencing and bioinformatic approaches are also considered. Structural vaccinology is an approach in which the detailed molecular structure of viral epitopes is used to design synthetic antigenic peptides. Virus-like particles are being designed for vaccine deliveries that are based on structures of viral capsid proteins and other synthetic lipopeptide building blocks. A new generation of adjuvants is being developed to further enhance immunogenicity, based on squalene and other oil–water emulsions, saponins, muramyl dipeptide, immunostimulatory oligonucleotides, Toll-like receptor ligands, and lymphotoxins. Finally, current trends in engineering of therapeutic antibodies including improvements of antigen-binding properties, pharmacokinetic and pharmaceutical properties, and reduction of immunogenicity are discussed. Taken together, understanding the chemistry of vaccine design, delivery and immunostimulation, and knowledge of the techniques of antibody design are allowing targeted development for the treatment of chronic disorders characterized by continuing activation of the immune system, such as autoimmune disorders, cancer, or allergies that have long been refractory to conventional approaches.
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Seagull, Robert W., Peter E. Lee, and Monica Frosch. "Comparison of microtubules and microfilaments in Tipula iridescent virus-infected and uninfected cells." Canadian Journal of Biochemistry and Cell Biology 63, no. 6 (June 1, 1985): 543–52. http://dx.doi.org/10.1139/o85-073.
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Microfilaments and microtubules were detected in Estigmene acrea virus-infected cells using fluorescent immunochemistry and in sections by electron microscopy. Twelve hours following infection of cells with Tipula iridescent virus, large virus assembly sites developed in the cytoplasm. The majority of infected cells exhibit no detectable changes in the cytoskeleton during the initial stage of infection, when virus assembly sites are forming. Actin was localized either in cytoplasmic spikes or in patches at the cell surface. Microtubules were parallel to the long axis of elongate cells or randomly distributed in globular cells. Intermediate filaments were not detected using either immunofluorscent or electron microscopic techniques. In later stages of infection some cells exhibit a specific association between actin and the virus assembly site. The significance of this observation remains unclear since only a portion of the population exhibits this change. From this study, it does not appear that cytoskeletal elements are of importance in the formation or maintenance of the membrane-free cytoplasmic virus assembly sites.
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Gutierrez-Garcia, Gonzalo, Teresa Cardesa, Luis Colomo, Fina Climent, Santiago Mercadal, Jose Luis Mate, Juan Manuel Sancho, et al. "Applicability of Different Immunohistochemistry Algorithms to Assess Gene Expression Profile In Patients with Diffuse Large B-Cell Lymphoma." Blood 116, no. 21 (November 19, 2010): 4134. http://dx.doi.org/10.1182/blood.v116.21.4134.4134.
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Abstract Abstract 4134 Gene expression profile (GEP) allows to distinguish two groups with different origin in patients with diffuse large B-cell lymphoma (DLBCL): germinal-center (GC) and activated (ABC), with the latter having a significantly poorer outcome. However, GEP is a technique not available in current clinical practice. For this reason, attempts to reproduce GEP data by immunophenotyping algorithms have been made. The aim of this study was to apply the most popular algorithms in a series of patients with DLBCL homogeneously treated with immunochemotherapy, in order to assess the correlation with GEP data and their usefulness to predict response and outcome of the patients. One hundred fifty seven patients (80M/77F; median age 65 years) diagnosed with DLBCL in 5 institutions of the Grup per l'Estudi dels Limfomes de Catalunya I Balears (GELCAB) during a 5-year period, treated with Rituximab-containing regimens (in most cases, R-CHOP), in whom histological material to construct a tissue microarrays (TMA) was available, constituted the subjects of the present study. Four algorithms were applied: Colomo (Blood 2003, 101:78) using CD10, bcl-6 and MUM1/IRF4; Hans (Blood 2004, 103:275) using CD10, bcl-6 and MUM1/IRF4; Muris (J Pathol 2006, 208:714) using CD10 and MUM1/IRF4, and Choi (Clin Cancer Res 2009, 15:5494), using CD10, bcl-6, GCET1, FOXP1 and MUM1/IRF4. The thresholds used were those previously described. GEP studies were performed in 62 patients in whom fresh frozen material was available. Main clinical and evolutive data were recorded and analyzed. The proportion of positive cases for the different single antigens was as follows: CD10 26%, bcl-6 64%, GCET1 46%, FOXP1 78% and MUM1/IRF4 28%. The distribution of cases (GC vs. non-GC) according to the algorithms is detailed in the table. In 88 of 110 patients (80%) with all the antigens available, the patients were allocated in the same group (either GC or non-GC). When the immunochemistry was compared with GEP data, the sensitivity in the GC group was 59%, 52%, 70% and 40% for Colomo, Hans, Muris and Choi algorithms, respectively. The sensitivity in the non-GC group was 81%, 85%, 62% and 84%, respectively. On the other hand, the positive predictive value (PPV) in the GC group was 81%, 83%, 72% and 77%, respectively. In non-GC subset the PPV for the different algorithms was 59%, 55%, 72% and 52%, respectively. We observed a higher percentage of misclassified cases in the GC-phenotype subset than in the non-GC subgroup. None of the immunohistochemical algorithms showed a significant superiority as surrogate of GEP information among the others. The ability of GEP groups as well as of groups defined by the algorithms to predict complete response (CR) rate, progression-free survival (PFS) and overall survival (OS) of the patients is showed in the table. Thus, whereas the GEP groups showed significant prognostic value for CR rate, PFS and OS, none of the immunohistochemical algorithms were able to predict the outcome. In conclusion, in a homogeneous series of DLBCL patients treated with immunochemotherapy, the different immunohistochemical algorithms were not able to mimic the GEP information. The prognostic impact of the groups defined by immunohistochemistry (GC vs. non-GC) was particularly low. N (%) CR rate N (%) 5-year PFS (%) 5-year OS (%) Colomo algorithm GC 53 (44) 39 (74) 48 54 Non-GC 68 (56) 53 (78) 55 62 Hans algorithm GC 61 (41) 47 (77) 54 60 Non-GC 88 (59) 67 (76) 52 59 Muris algorithm GC 87 (57) 63 (72) 48 57 Non-GC 65 (43) 51 (78) 56 63 Choi algorithm GC 45 (33) 32 (71) 48 54 Non-GC 90 (67) 70 (78) 52 61 Gene expression profile 30 (58) 25 (83) 76* 80** GC Activated 22 (42) 17 (77) 31* 45** * p=0.005, ** p=0.03. Disclosures: No relevant conflicts of interest to declare.
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Fleurot, Isabelle, Marina Aigle, Renaud Fleurot, Claire Darrigo, Jacques-Antoine Hennekinne, Alexandra Gruss, Elise Borezée-Durant, and Agnès Delacroix-Buchet. "Following Pathogen Development and Gene Expression in a Food Ecosystem: the Case of a Staphylococcus aureus Isolate in Cheese." Applied and Environmental Microbiology 80, no. 16 (June 13, 2014): 5106–15. http://dx.doi.org/10.1128/aem.01042-14.
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ABSTRACTHuman intoxication or infection due to bacterial food contamination constitutes an economic challenge and a public health problem. Information on thein situdistribution and expression of pathogens responsible for this risk is to date lacking, largely because of technical bottlenecks in detecting signals from minority bacterial populations within a complex microbial and physicochemical ecosystem. We simulated the contamination of a real high-risk cheese with a natural food isolate ofStaphylococcus aureus, an enterotoxin-producing pathogen responsible for food poisoning. To overcome the problem of a detection limit in a solid matrix, we chose to work with a fluorescent reporter (superfolder green fluorescent protein) that would allow spatiotemporal monitoring ofS. aureuspopulations and targeted gene expression. The combination of complementary techniques revealed thatS. aureuslocalizes preferentially on the cheese surface during ripening. Immunochemistry and confocal laser scanning microscopy enabled us to visualize, in a single image, dairy bacteria and pathogen populations, virulence gene expression, and the toxin produced. This procedure is readily applicable to other genes of interest, other bacteria, and different types of food matrices.
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Naryzhny, S. N., N. L. Ronzhina, M. A. Mainskova, N. V. Belyakova, R. A. Pantina, and M. V. Filatov. "Development of barcode and proteome profiling of glioblastoma." Biomeditsinskaya Khimiya 60, no. 3 (2014): 308–21. http://dx.doi.org/10.18097/pbmc20146003308.
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High grade glioma (glioblastoma) is the most common brain tumor. Its malignancy makes it the fourth biggest cause of cancer death. In our experiments we used several glioblastoma cell lines generated in our laboratory to obtain proteomics information specific for this disease. This study starts our developing the complete 2DE map of glioblastoma proteins. 2DE separation with following imaging, immunochemistry, spot picking, and mass-spectrometry allowed us detecting and identifying more than 100 proteins. Several of them have prominent differences in their level between norm and cancer. Among them are alpha-enolase (ENOA_HUMAN), pyruvate kinase isozymes M1/M2 (KPYM_HUMAN), cofilin 1 (COF1_HUMAN), translationally-controlled tumor protein TCTP_HUMAN, annexin 1 (ANXA1_HUMAN), PCNA (PCNA_HUMAN), p53 (TP53_HUMAN) and others. Most interesting results were obtained with protein p53. In all glioblastoma cell lines, its level was dramatically up regulated and enriched by multiple additional isoforms. This distribution is well correlated with presence of these proteins inside of cells themselves. At this initial step we suggest the panel of specific brain tumor markers (signature) to help creating noninvasive techniques to diagnose disease. These preliminary data point to these proteins as promising markers of glioblastoma.
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Botto, Barbara, Domenico Novero, Annalisa Chiappella, Mattia Novo, Alessia Castellino, Chiara Ciochetto, Giovanni Cametti, et al. "The Prognostic Value of MYC, BCL2 and BCL6 Overexpression Evaluated By Immunohistochemistry (IHC) in De-Novo Diffuse Large B Cell Lymphoma (DLBCL) Treated with Rituximab-CHOP." Blood 124, no. 21 (December 6, 2014): 2964. http://dx.doi.org/10.1182/blood.v124.21.2964.2964.
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Abstract Introduction : MYC, BCL2 and BCL6 overexpression, assessed by IHC, with the latter conferring a better prognosis, have been reported to be a prognostic factor in DLBCL, but data are not consistent and sometimes contradictory. The aim of the present study was to assess the prognostic impact of overexpression of MYC, BCL2, and BCL6 in a retrospective cohort of de-novo DLBCL, selected for an high proliferation index (MIB1 ≥70%), treated consecutively with R-CHOP regimen. Methods: Patients with de-novo DLBCL diagnosed between January 2010 and December 2013 were included into the study. Inclusion criteria were: high proliferation index MIB1 ≥ 70% and a full course of R-CHOP regimen. Paraffin-embedded tumor samples were collected and investigated using immunohistochemistry (IHC) for MYC, BCL2 and BCL6. Fluorescence in situ hybridization (FISH) is ongoing. MYC/BCL2+ or MYC/BCL6+ double expression cases were identified if they had rearrangements of MYC and BCL2 or BCL6. MYC immunochemistry was done on TMA sections using the antibody clone Y69. BCL2 and BCL6 staining had been evaluated previously at diagnosis. Tumor cells were defined positive for MYC and BCL2 or BCL6 protein expression by immunostaining if >40%, >40% and >25% of cells showed positive expression, respectively. Progression free survival curves (PFS) were estimated using the Kaplan-Meier method and compared between groups using the log-rank test and Cox models. Results : One hundred and sixty seven patients are evaluable for clinical characteristics and 69/167 had paraffin embedded tumor samples available for immunohistochemistry at the time of present analysis. Clinical characteristics of the 69 cases were: median age 66 years (IQR 57;73), 45 (65%) male, 47 (68%) stage III-IV, 35 (54%) with elevated LDH levels and 46 (67%) at International Prognostic Index (IPI) high intermediate or high risk. Overexpression of MYC was detected in 28 cases (41%), 50 (72%) and 38 (55%) showed BCL2 and BCL6 overexpression respectively. Nineteen (28%) cases showed MYC/BCL2+ and 17 (25%) MYC/BCL6+ double expression. With a median follow up of 26 months, the median 2-years PFS was 59%. Overexpression of MYC and BCL2 proteins and low expression of BCL6 were associated with an inferior 2-years PFS in univariate analysis: MYC- vs MYC+ 64% vs 55%; BCL2- vs BCL2+ 71% vs 56%; BCL6+ vs BCL6- 61% vs 54%. In a Cox multivariate regression model adjusted for IPI and age, MYC overexpression, BCL2 positivity and BCL6 negativity showed prognostic relevance as significant independent indicators with different risk (Hazard ratio 2.53 for MYC+, 2.08 for BCL2+ and 1.62 for BCL6-). Established that the three variable contributed with different risk in the multivariate analysis, an IHC sum additive score of 0-5 was calculated proportionally to the coefficient estimated (coefficient [Log hazard ratio] 0.92 for MYC+, 0.73 for BCL2+ and 0.48 for BCL6-), assigning an individual risk of 2 points for MYC or BCL2 positivity and 1 point for BCL6 negativity. Two years-PFS was significantly different between all separate groups (Hazard ratio for unit increase 1.57 95% CI 1.11-2.22, p=0.01). After pooling scores 0-1 (with or without BCL6), 2 (presence of MYC or BCL2 only), and 3-4-5 (MYC+/BCL6-, BCL2+/BCL6-, MYC+/BCL2+, MYC+/BCL2+/BCL6-) 2-yrs PFS rates were different across the three groups: 100% vs 64% vs 50% (log rank p= 0.04) (figure 1). Conclusion: Our data showed, with the limits of a small sample size, that MYC overexpression alone or with high expression of BCL2 and/or low expression of BCL6 correlates with a worse prognosis independently by IPI score in a cohort of DLBCL selected for high proliferation index and treated with R-CHOP. Assessment of MYC, BCL2 and BCL6 expression by IHC represents a rapid, inexpensive, and reproducible technique. These results need to be confirmed in our complete series of 167 patients (analysis ongoing) and validated prospectively in a larger cohort, using standardized staining and scoring methodologies. Thus, MYC and BCL2 represent relevant biomarkers that should be tested in future clinical trials using novel effective and targeted agents in order to improve the prognosis of DLBCL. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.
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Skoupilova, Hana, Vladimir Rak, Jiri Pinkas, Jindrich Karban, and Roman Hrstka. "The Cytotoxic Effect of Newly Synthesized Ferrocenes against Cervical Carcinoma Cells Alone and in Combination with Radiotherapy." Applied Sciences 10, no. 11 (May 28, 2020): 3728. http://dx.doi.org/10.3390/app10113728.
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Cervical cancer is one of the most common types of cancer in women, with approximately 500,000 new cases and 250,000 deaths every year. Radiotherapy combined with chemotherapy represents the treatment of choice for advanced cervical carcinomas. The role of the chemotherapy is to increase the sensitivity of the cancer cells to irradiation. Cisplatin, the most commonly used drug for this purpose, has its limitations. Thus, we used a family of ferrocene derivatives (in addition, one new species was prepared using standard Schlenk techniques) and studied their effects on cervical cancer cells alone and in combination with irradiation. We applied colorimetric assay to determine the cytotoxicity of the compounds; flow cytometry to analyze the production of reactive oxygen species (ROS), cell cycle, and mitochondrial membrane potential (MMP); immunochemistry to study protein expression; and colony forming assay to evaluate changes in radiosensitivity. Treatment with ferrocenes exhibited significant cytotoxicity against cervical cancer cells, associated with increasing ROS production and MMP changes, suggesting the induction of apoptosis. The combined activity of ferrocenes and ionizing radiation highlighted ferrocenes as potential radiosensitizing drugs, while their higher single-agent toxicity in comparison with routinely used cisplatin could also be promising. Our results demonstrate antitumor activity of several tested ferrocenes both alone and in combination with radiotherapy.
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Wilkins, T. A., G. Brouwers, J. C. Mareschal, and C. L. Cambiaso. "High sensitivity, homogeneous particle-based immunoassay for thyrotropin (Multipact)." Clinical Chemistry 34, no. 9 (September 1, 1988): 1749–52. http://dx.doi.org/10.1093/clinchem/34.9.1745.
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Abstract We describe the first homogeneous, nonradioactive, high-sensitivity assay for human thyrotropin (TSH). The assay is based on particle immunoassay techniques, wherein 800-nm particles form the basis for the immunochemistry, delivery, and the detection technologies, respectively. Our assay also is the first to involve the use of fragmented monoclonal antibodies (to eliminate serum interferences) covalently coupled to particles without loss of their binding properties. Assays are performed in a semiautomated mode with use of a new modular system (Multipact). Equilibrium is reached in less than 2 h. Precision profile, sensitivity, and clinical studies indicate that the assay is accurate, has good precision at low concentrations, and that detection-limit characteristics compare well with those of a leading commercial high-sensitivity immunoradiometric assay (IRMA) for TSH. Dilution characteristics were satisfactory down to the assay's detection limit for a range of clinical samples. Correlation studies vs a reference IRMA method yielded the regression equation, present method = 0.976 (IRMA) + 0.002 milli-int. unit/L (r = 0.98), for 223 samples with TSH concentrations in the range 0 to 30 milli-int. units/L. For 40 samples with TSH less than or equal to 1.0 milli-int. unit/L it was: present method = 0.94 (IRMA) + 0.005 milli-int. unit/L (r = 0.96).
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Jing, Zhen, Changzheng Shi, Lihui Zhu, Yonghui Xiang, Peihao Chen, Zhilin Xiong, Wenxian Li, Yiwen Ruan, and Li'an Huang. "Chronic Cerebral Hypoperfusion Induces Vascular Plasticity and Hemodynamics but Also Neuronal Degeneration and Cognitive Impairment." Journal of Cerebral Blood Flow & Metabolism 35, no. 8 (April 8, 2015): 1249–59. http://dx.doi.org/10.1038/jcbfm.2015.55.
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Chronic cerebral hypoperfusion (CCH) induces cognitive impairment, but the compensative mechanism of cerebral blood flow (CBF) is not fully understood. The present study mainly investigated dynamic changes in CBF, angiogenesis, and cellular pathology in the cortex, the striatum, and the cerebellum, and also studied cognitive impairment of rats induced by bilateral common carotid artery occlusion (BCCAO). Magnetic resonance imaging (MRI) techniques, immunochemistry, and Morris water maze were employed to the study. The CBF of the cortex, striatum, and cerebellum dramatically decreased after right common carotid artery occlusion (RCCAO), and remained lower level at 2 weeks after BCCAO. It returned to the sham level from 3 to 6 weeks companied by the dilation of vertebral arteries after BCCAO. The number of microvessels declined at 2, 3, and 4 weeks but increased at 6 weeks after BCCAO. Neuronal degeneration occurred in the cortex and striatum from 2 to 6 weeks, but the number of glial cells dramatically increased at 4 weeks after BCCAO. Cognitive impairment of ischemic rats was directly related to ischemic duration. Our results suggest that CCH induces a compensative mechanism attempting to maintain optimal CBF to the brain. However, this limited compensation cannot prevent neuronal loss and cognitive impairment after permanent ischemia.
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Emerson, Jane F., Gilda Ngo, and Scott S. Emerson. "Screening for Interference in Immunoassays." Clinical Chemistry 49, no. 7 (July 1, 2003): 1163–69. http://dx.doi.org/10.1373/49.7.1163.
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Abstract Background: The presence of interfering substances in patient samples submitted for immunoassay cannot be reliably anticipated. We therefore evaluated three interference screening techniques and estimated the prevalence of interfering substances as defined by positive outcomes with these protocols. Methods: We evaluated 160 samples for the presence of substances that may interfere with four immunoassays (40 samples for each): thyroid-stimulating hormone, prostate-specific antigen, β-human chorionic gonadotropin, and cortisol. Interference was defined by nonlinear responses with serial dilution, discrepant results after pretreatment with heterophile blocking reagent (HBR), and positive reactions on a mouse-antibody-negative control reaction (Tandem ICON® ImmunoConcentration HCG). Criteria for declaring significant discrepant results were based on a Z-score computed using the assay CV. The McNemar test was used to compare the prevalence of discrepancies across the three screening techniques. The association between type of immunoassay and prevalence of discrepant results was determined by a modified Pearson χ2 statistic. Results: Five of the 160 samples [3.1%; 95% confidence interval (CI), 1.0–7.1%] screened positive with the ICON. Seventy-two of the 148 samples with informative serial dilutions (48.6%; 95% CI, 40.4–57.0%) had at least one discrepant result at higher dilutions. After pretreatment with HBR, 53 of the 140 samples (38%; 95% CI, 29.8–46%) were discrepant. Only 48 of the 140 samples with informative measurements for all three screening techniques (34%; 95% CI, 26–43%) were negative by all three. The prevalence of positive screens varied significantly by type of immunoassay (P <0.0001) for both HBR and serial dilution. Only 3% (0.8–7%) of the samples tested with HBR showed a change from normal to abnormal or the reverse after treatment. Conclusions: Introducing a protocol based on any of these three techniques into the immunochemistry laboratory to prescreen for interfering substances is not warranted. The evaluation of specimens for the presence of interfering anti-animal antibodies should be reserved for cases in which clinical history or suspicious results indicate the need.
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Mandel, Irwin D. "A Contemporary View of Salivary Research." Critical Reviews in Oral Biology & Medicine 4, no. 3 (April 1993): 599–604. http://dx.doi.org/10.1177/10454411930040034701.
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The past 50 years of salivary research has been marked by a series of changing perceptions as new techniques and technologies have illuminated the complexities of the secretory mechanism, salivary composition, and function. The modem era began with the innovations of electrophoresis, chromatography, histochemistry, immunochemistry, electron microscopy, and microphysiology. The idea of saliva as primarily a digestive fluid composed of salts, amylase, and mucin was rapidly broadened to encompass a wide spectrum of protective proteins with the dual responsibility of protecting both hard and soft tissues. Characterization of the secretory IgA and nonimmunological antibacterial systems and the proteins responsible for the regulation of calcium and phosphate levels dominated the research in the 1960s and 1970s. An appreciation of the nature, formation, and role of the salivary pellicle and the interplay between bacterial adherence and agglutination provided a clinical thrust. Morphologists and physiologists redefined the secretory process on a molecular level. The 1980s saw the union of structure and function, both in terms of synthesis and release of the secretory products and their specific roles in the oral cavity in health and disease. The excitement of the 1990s is in the genetic control of processes and products, elucidating the mechanisms, and using the information to improve on nature: an era of great expectations and hubris. This article is essentially a personal guided tour through the past 50 years of salivary research.
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Wieczorek, D. F., M. Periasamy, G. S. Butler-Browne, R. G. Whalen, and B. Nadal-Ginard. "Co-expression of multiple myosin heavy chain genes, in addition to a tissue-specific one, in extraocular musculature." Journal of Cell Biology 101, no. 2 (August 1, 1985): 618–29. http://dx.doi.org/10.1083/jcb.101.2.618.
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We have investigated the developmental transitions of myosin heavy chain (MHC) gene expression in the rat extraocular musculature (EOM) at the mRNA level using S1-nuclease mapping techniques and at the protein level by polypeptide mapping and immunochemistry. We have isolated a genomic clone, designated lambda 10B3, corresponding to an MHC gene which is expressed in the EOM fibers (recti and oblique muscles) of the adult rat but not in hind limb muscles. Using cDNA and genomic probes for MHC genes expressed in skeletal (embryonic, neonatal, fast oxidative, fast glycolytic, and slow/cardiac beta-MHC), cardiac (alpha-MHC), and EOM (lambda 10B3) muscles, we demonstrate the concomitant expression at the mRNA level of at least six different MHC genes in adult EOM. Protein and immunochemical analyses confirm the presence of at least four different MHC types in EOM. Immunocytochemistry demonstrates that different myosin isozymes tend to segregate into individual myofibers, although some fibers seem to contain more than one MHC type. The results also show that the EOM fibers exhibit multiple patterns of MHC gene regulation. One set of fibers undergoes a sequence of isoform transitions similar to the one described for limb skeletal muscles, whereas other EOM myofiber populations arrest the MHC transition at the embryonic, neonatal/adult, or adult EOM-specific stage. Thus, the MHC gene family is not under the control of a strict developmental clock, but the individual genes can modify their expression by tissue-specific and/or environmental factors.
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Ivovic, Miomira, Vladan Zivaljevic, Svetlana Vujovic, Ljiljana Marina, Milina Tancic-Gajic, Dusan Dundjerovic, Marija Barac, and Dragan Micic. "Small bowel adenocarcinoma mimicking a large adrenal tumor." Srpski arhiv za celokupno lekarstvo 141, no. 3-4 (2013): 232–36. http://dx.doi.org/10.2298/sarh1304232i.
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Introduction. Adenocarcinoma of the small bowel is a rare gastrointestinal neoplasm usually affecting the distal duodenum and proximal jejunum. Because of their rarity and poorly defined abdominal symptoms, a correct diagnosis is often delayed. Case Outline. We present a 43-year-old woman admitted at the Clinic for Endocrinology due to a large tumor (over 7 cm) of the left adrenal gland. The tumor was detected by ultrasound and confirmed by CT scan. The patient complained of abdominal pain in the left upper quadrant, fatigue and septic fever. Normal urinary catecholamines excluded pheochromocytoma. The endocrine evaluations revealed laboratory signs of subclinical hypercorticism: midnight cortisol 235 nmol/L, post 1 mg - overnight Dexamethasone suppression test for cortisol 95.5 nmol/L and basal ACTH 4.2 pg/mL. Plasma rennin activity and aldosterone were within the normal range. Surgery was performed. Intraoperative findings showed signs of acute peritonitis and a small ulceration of the jejunum below at 70 cm on the anal side from the Treitz?s ligament. Adrenal glands were not enlarged. Patohistology and immunochemistry identified adenocarcinoma of the jejunum without infiltration of the lymphatic nodules. The extensive jejunal resection and lavage of the peritoneum were performed. Due to complications of massive peritonitis, the patient died seven days after surgery. Conclusion. Poorly defined symptoms and a low incidence make the diagnosis of small bowel carcinoma, particularly of the jejunal region, very difficult in spite of the new endoscopic techniques.
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Grogan, TM, BG Durie, C. Lomen, C. Spier, DP Wirt, R. Nagle, GS Wilson, L. Richter, E. Vela, and V. Maxey. "Delineation of a novel pre-B cell component in plasma cell myeloma: immunochemical, immunophenotypic, genotypic, cytologic, cell culture, and kinetic features." Blood 70, no. 4 (October 1, 1987): 932–42. http://dx.doi.org/10.1182/blood.v70.4.932.932.
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Abstract A novel pre-B cell component in direct and cultured myeloma bone marrow material has been delineated by using immunochemistry and flow cytometry techniques. Our phenotypic studies suggest a novel hybrid expression of pre-B and plasma cell antigens with coexpression of cytoplasmic mu, common acute lymphoblastic leukemia antigen, terminal deoxynucleotidyl transferase, and plasma cell antigens (PCA-1 and PC- 1). This suggests that myeloma pre-B-like cells are aberrant malignant cells and not normal pre-B lymphocytic counterparts. With the advantage of a pure and stable source of these cells from M3 culture to allow molecular characterization, we performed one- and two-dimensional gel electrophoresis and Western blotting. We found that the cytoplasmic mu in myeloma pre-B-like cells has a molecular weight of 74,000 daltons and an isoelectric point of 6.3 and that it is strikingly homogeneous and discrete in size and charge compared with standard secretory mu, which suggests an aberrant, mutant, or monoclonal form of mu. Monoclonality was further evidenced by heavy- and light-chain immunoglobulin gene rearrangements demonstrated with JH and C kappa probes. We also established that this novel myeloma pre-B component is a major proliferative element as determined by double-labeling experiments with phenotype coupled to labeling/proliferative indexes. Our stimulatory studies indicate some capacity of these cells to mature on exposure to phorbol esters. These myeloma pre-B cells may represent the stem cell or self-renewal component in myeloma. Our establishment of these cells in long-term culture offers a considerable asset in studying the immature cells, which may be critical to the immortalization of myeloma.
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Grogan, TM, BG Durie, C. Lomen, C. Spier, DP Wirt, R. Nagle, GS Wilson, L. Richter, E. Vela, and V. Maxey. "Delineation of a novel pre-B cell component in plasma cell myeloma: immunochemical, immunophenotypic, genotypic, cytologic, cell culture, and kinetic features." Blood 70, no. 4 (October 1, 1987): 932–42. http://dx.doi.org/10.1182/blood.v70.4.932.bloodjournal704932.
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A novel pre-B cell component in direct and cultured myeloma bone marrow material has been delineated by using immunochemistry and flow cytometry techniques. Our phenotypic studies suggest a novel hybrid expression of pre-B and plasma cell antigens with coexpression of cytoplasmic mu, common acute lymphoblastic leukemia antigen, terminal deoxynucleotidyl transferase, and plasma cell antigens (PCA-1 and PC- 1). This suggests that myeloma pre-B-like cells are aberrant malignant cells and not normal pre-B lymphocytic counterparts. With the advantage of a pure and stable source of these cells from M3 culture to allow molecular characterization, we performed one- and two-dimensional gel electrophoresis and Western blotting. We found that the cytoplasmic mu in myeloma pre-B-like cells has a molecular weight of 74,000 daltons and an isoelectric point of 6.3 and that it is strikingly homogeneous and discrete in size and charge compared with standard secretory mu, which suggests an aberrant, mutant, or monoclonal form of mu. Monoclonality was further evidenced by heavy- and light-chain immunoglobulin gene rearrangements demonstrated with JH and C kappa probes. We also established that this novel myeloma pre-B component is a major proliferative element as determined by double-labeling experiments with phenotype coupled to labeling/proliferative indexes. Our stimulatory studies indicate some capacity of these cells to mature on exposure to phorbol esters. These myeloma pre-B cells may represent the stem cell or self-renewal component in myeloma. Our establishment of these cells in long-term culture offers a considerable asset in studying the immature cells, which may be critical to the immortalization of myeloma.
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Giraldo, Pilar, Paz Latre, Esther Franco-García, Ramiro Alvarez, Mar Pascual, Tomas Castilla, Milena Sants, and Carmen Martos. "HAEMACARE Project: Reclassifing Hematologic Neoplasias According Morphological Data." Blood 110, no. 11 (November 16, 2007): 4392. http://dx.doi.org/10.1182/blood.v110.11.4392.4392.
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Abstract Aims: The European Project HAEMACARE (VI Frame Research Program), is working in the reclassification of hematological neoplasias included in population based Cancer Registries of 13 European countries with the objective to improve and standardize morphological data validation. Patients and methods: a multidisciplinary research group of the I+CS integrated by epidemiologists, hematologists, pathologists is working in the reclassification of Hodgkin disease (HD) included in the population based Cancer Registry of Zaragoza (CRZ) in order to reclassify according to WHO and ICD-O classifications and identify the non other specification (NOS) cases. From January 1995 to December 2000, a total of 141 patients diagnosed as Hodgkin disease are included. The sources of data were: Zaragoza population based Cancer Registry, Aragon hematological malignancies Registry (FEHHA) and from the records of public Hospitals and the National Deaths Index. All cases were identified from the CRZ and 35 cases (24.8%) NOS were re-studied by the hematologists and pathologists and reclassified according histological and morphological subtypes of WHO classification. Diagnostic criteria were set for LRCHD: scattered Hodgkin-Reed-Sternberg cells with a classical immunophenotype in a background of small lymphocytes without admixture of eosinophils and neutrophils and without sclerosis nodular lymphocyte predominant HD (NLPHD), mixed cellular (MCD) and lymphocyte-rich classical HD (LRCHD). In 24.8% of cases were necessary to process the samples again or review smears and applied immunochemistry techniques in order to establish accurate diagnosis according the new sub-classification in NLPHD or CHD and distinguish some cases of T-cell rich large B-cell lymphoma. In addition we have calculated the estimated survival according specific morphological subtypes. Results: Hodgkin Disease. Histological distribution ICD-0-3 No % males females mean age(y) mean surv(y) No deaths (%) *non other specification NOS* 35 24.8 15 20 43.7 5.8 13 (34.3) CHD 64 45.4 39 25 39.3 7.6 19 (29.7) MCD 21 14.9 15 6 46.3 7.7 4 (19.0) LRCHD 11 7.8 9 2 51.1 6.7 5 (45.5) NLPHD 10 7.0 6 4 51.2 8.3 2 (20.0) Total 141 84 57 43.2 7.1 43 (30.5) The estimated survival analysis showed a mean of 7.6 years (95% CI: 6.7–8.4) for all cases, median not reached. Comments: In our study has been necessary to reclassify the 24.8% of cases according WHO criteria. The incidence peaks are classical. The overall survival is similar to describe previously.
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Rivas-Vera, Silvia, Mabel Oropeza-Borges, and Pedro Sobrevilla-Calvo. "T- Cell Non-Hodgkin's Lymphoma, Data From a Mexican Tertiary Health Center.." Blood 114, no. 22 (November 20, 2009): 5030. http://dx.doi.org/10.1182/blood.v114.22.5030.5030.
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Abstract Abstract 5030 Introduction T cell Non/Hodgkin's lymphomas (TCNHL) are a group of lymphomas characterized by an aggressive clinical course and resistance to the usual chemotherapy agents. Due to the limited availability of immunochemistry techniques the frequency and clinical presentation of these lymphomas are not well described in underdeveloped countries. Here we describe the experience with these diseases in a tertiary referral health center. Patients and methods We reviewed 520 cases with diagnosis of Non-Hodgkin's Lymphoma from the records of the Pathology Department of the Hospital General de Mexico, seen from January 2002 to July 2006. Results We found 80 cases with TCNHL (15.3%). In 62 cases the clinical information was available for review. We found a 1.45:1 male/female ratio, median age 32 years (range 17 to 83). Symptoms were present for a median of 6 months (range 1 to 120 months) before diagnosis. Regarding the pathological classification the Unspecified Peripheral T-cell lymphomas were more common (56%), followed by the anaplastic lymphoma (23%), T/NK (16%), Cutaneous lymphoma (3%) and angioimmunoblastic lymphoma (2%). In 56% the initial presentation was nodal and in 44% extranodal. The extranodal sites were: Nasal (34%), bone marrow (22%), pleura and lung (14%), parotid (5%), colon (5%), and liver (5%). Sixty seven percent of the patients had advanced clinical stages and 73% had at least 3 B symptoms. One quarter of patients had bulky disease at their first visit to our Hospital; the most frequent site was retroperitoneal (19%). The lactic dehydrogenase (LDH) was elevated in 89% of cases (range:97 U/L -3017 IU L, median 418 U/L); patients with anaplastic NHL had the highest values and NHL-TNK patients the lowest. HIV serology was performed in 41 of 62 patients. Ninety four percent of the patients had an ECOG between 1 and 3. We calculated the International Prognostic Index (IPI); it was low in 23 patients (37%), low-intermediate in 18 (29%), intermediate-high in 16 (26%) and high in just 5 patients (8%). When we classified the patients with the Prognostic Index for non-specific peripheral T-NHL (PIT or PTCL-L), we found that 79.4% of cases were in the high-risk group in contrast with 73.6% low group according to IPI. All patients were treated with CHOP chemotherapy. A complete response (CR) was achieved in 10 patients (16%); 13 patients (21%) were still on treatment at the end of the study, 4 patients (6.45%) did not respond to treatment and 14 patients (24.1%) had progressive disease. Ten patients discontinued treatment (16%) and 5 died (8.06%). At the end of the observation period 8 patients were alive without tumor activity, 2 patients were lost without tumor activity, 7 patients were alive with disease, 24 patients were missing with tumor activity, 9 patients were on treatment and 12 patients had died. Conclusions This study shows that in Mexico as in other populations, the TCNHL are less frequent than the B-cell lymphomas. It also confirms the poor response of the T-Cell lymphomas to CHOP. The population that attends our hospital is of a low socioeconomic stratum, and this fact explains the large abandonment of medical care. It is desirable the development of new drugs for these neoplasia. Disclosures No relevant conflicts of interest to declare.
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Kanniah, Umamaheswari, Unnimaya Varghese, V. S. Sruthi, and Mathew Jacob. "Immunofluorescence in Oral Dermatological Disorders- No Shiny Matter." Journal of Academy of Dental Education, May 29, 2017, 24. http://dx.doi.org/10.18311/jade/0/15951.
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Dermatological diseases with underlying immunological pathosis exhibiting similar clinical manifestations have challenged even the veteran dermatologists in rendering diagnosis clinically, necessitating study of the skin lesions with immunofluroscence. The technique is similar to immunochemistry where instead of enzymes antibodies are labelled with a fluorescent dye4. It probes the study of the cell and molecular biology. Immunofluorescence permit early diagnosis, treatment and subsequent monitoring of disease activity in patient with potentially life threatening disease. It is widely used in the field of vesiculo-bullous lesions and other related oral dermatological disorders by demonstrating antibodies either in biopsy tissue or in the serum separated from blood collected from the patients.
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Lesieur, Emmanuelle, Julia Torrents, Frédéric Fina, Christine Zandotti, Julie Blanc, Sophie Collardeau-Frachon, Céline Gazin, et al. "Congenital infection of SARS-CoV-2 with intrauterine foetal death: a clinicopathological study with molecular analysis." Clinical Infectious Diseases, September 23, 2021. http://dx.doi.org/10.1093/cid/ciab840.
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Abstract Observations of vertical transmission of SARS-CoV-2 infection from mother to foetus have recently been described in the literature. However, the consequences of such transmission, whether foetal or neonatal, are poorly understood. From a case of in utero foetal death at 24 +2 weeks of gestation that occurred seven days after the diagnosis of symptomatic SARS-CoV-2 infection in the mother, we isolated the incriminating virus by immunochemistry and molecular techniques in several foetal tissues, with a variant analysis of the SARS-CoV-2 genome. Moreover, the foetal demise could be explained by the presence of placental histological lesions, such as histiocytic intervillositis and trophoblastic necrosis, in addition to foetal tissue damage. We observed mild foetal growth retardation and visceral damage to the liver, causing hepatocellular damage and haemosiderosis. To the best of our knowledge, this is the first report in the literature of foetal demise secondary to maternal-foetal transmission of SARS-CoV-2 with a congenital infection and a pathological description of placental and foetal tissue damage. SARS-CoV-2 was identified in both specimens by three independent techniques (immunochemistry, RT-qPCR and RT-dPCR). Furthermore, the incriminating variant has been identified.
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"Nano-TiO2 Photodynamic Treatment on Cancer Cells Combining with Immunochemistry and Electroporation Techniques." ECS Meeting Abstracts, 2007. http://dx.doi.org/10.1149/ma2007-02/12/810.
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