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51.
Claudio Gustavo Stefanoff Rocío Hassan Ana Carolina Gonzalez Luiz Ant?nio B Andrade Daniel G Tabak Sérgio Romano Ilana R Zalcberg 《Diagnostic molecular pathology》2003,12(2):79-87
We herein present a technical strategy to optimize DNA isolation from paraffin-embedded tissue (PET). This includes the choice of adequate buffers for proteinase K digestion and multiplex PCR amplifications for assessing the appropriateness of DNA extracts for subsequent PCR assays for detecting clonality. We found that the association of proteinase K digestion in nonionic buffer and subsequent extract dilutions accounted for 79% of successful amplifications. A final efficiency of 88% was achieved by additional organic extractions and/or re-extractions. Comparisons were carried out with control DNA extracts from fresh samples to assess the efficiency of each clonality assay. Immunoglobulin CDRIII rearranged region amplification was more efficient for pregerminal center B-cell lymphomas in contrast to CDRII rearrangement detection, which was more effective for germinal and postgerminal lymphomas. T-cell clonality detection by TCRgamma PCR was less efficient in PET samples than in fresh tissues showing that DNA integrity is more critical for TCR than for IGH amplification. Two inconclusive cases without phenotypic markers and two other atypical lymphoproliferations masked by reactive T cells were diagnosed as plasmablastic lymphomas and as monoclonal B-proliferations, respectively, due to IGH rearrangements. 相似文献
52.
53.
Karina R B Bastos Renato Barboza Rosa M Elias Luiz R Sardinha Marcos G Grisotto Cláudio R F Marinho Gustavo P Amarante-Mendes José M Alvarez Maria Regina D'Império Lima 《Journal of interferon & cytokine research》2002,22(12):1191-1199
Aiming to clarify the role of endogenous interleukin-12 (IL-12) in protective immunity against blood stages of Plasmodium chabaudi chabaudi (AS), we evaluated the course of infection in IL-12p40 gene knockout (IL-12p40KO) and wild-type (WT) C57BL/6 mice, focusing (1) on the ability of T cells to develop adequate type 1 responses and (2) on the potentiality of macrophages to respond to parasites, interferon-gamma (IFN-gamma), or both. We observed that IL-12p40KO mice develop significantly higher parasitemias during the acute infection, although mice from both groups clear the parasites within a month and similarly eliminate a secondary challenge. Thus, fully protective immunity to P. c. chabaudi can be generated in the absence of IL-12. However, this cytokine may promote parasite control during the early phase of infection. The increased acute parasitemia of IL-12p40KO mice was associated with both impaired IFN-gamma and nitric oxide (NO) response by spleen cells. Because stimulation with recombinant IFN-gamma (rIFN-gamma) failed to improve the NO response in IL-12p40KO macrophages, we investigated whether these cells have an intrinsic defect. Analysis of peritoneal macrophages revealed that IL-12p40KO cells produce higher levels of transforming growth factor-beta1 (TGF-beta1) compared with WT cells and respond to infected erythrocytes or rIFN-gamma by releasing little NO. Moreover, IL-12p40KO macrophages had a severely impaired ability to internalize opsonized infected erythrocytes, suggesting that the low effector profile assumed by these cells may compromise antibody-mediated immunity. Taken together, our results support the idea that the absence of IL-12p40 not only affects IFN-gamma production but also has deep consequences in macrophage effector functions that may contribute to exacerbation of the early phase of P. c. chabaudi malaria. 相似文献
54.
1. The horizontal and vertical vestibuloocular reflex (HVOR and VVOR, respectively) was studied in four chronically instrumented pigeons. Eye movements were measured using the magnetic search-coil technique and were produced by rotation in the dark. During the rotation paradigms, the pigeons were either pharmacologically aroused (using amphetamine) or drug free (normal). The pigeon HVOR and VVOR were tested using step and sinusoidal rotational stimulation. The range of frequencies (0.03-6.0 Hz) and the magnitude of the sinusoidal stimuli were chosen to match those used in a previous study of the responses of semicircular canal primary afferents (SCPAs) in unanesthetized (i.e., normal) pigeons. 2. The gain of the HVOR and VVOR in both normal and aroused pigeons was independent of stimulus magnitude (6-30 degrees/s) over the frequency range tested. In aroused pigeons, the frequency independent gain (G) of the HVOR (G = 0.6) and VVOR (G = 0.9) was roughly twice that for normal pigeons. Pigeon VOR phase under all combinations of orientation and arousal level was independent of stimulus magnitude except at the lowest frequency tested (0.03 Hz). At this frequency, phase lead decreased as stimulus magnitude increased for the HVOR and VVOR in both normal and aroused pigeons. 3. The step and sinusoidal gains were greater for the VVOR than for the HVOR under the same level of arousal. Neither the gain nor the dominant time constant of the VOR (tau vor) differed for rotation direction (clockwise or counterclockwise) for the HVOR or VVOR in normal or aroused pigeons. 4. The mean value of tau vor was 4.0 +/- 0.5 (SE) s as estimated from frequency response data and 4.3 +/- 0.4 s as estimated from step response data for the HVOR and VVOR in both normal and aroused pigeons. In comparison, the mean value of the dominant or cupular time constant (tau c) of normal pigeon SCPAs was approximately 10 s as estimated from frequency response data. These results indicate that tau vor is shorter than tau c in the pigeon. 5. At higher frequencies, the pigeon HVOR and VVOR exhibit an increasing phase lag unaccompanied by a gain change--characteristics produced by a pure time delay. The value of this time delay was about 7 ms for both the HVOR and VVOR in both normal and aroused pigeons. The HVOR and VVOR in normal and aroused pigeons lacked the higher frequency lead characteristics present in the frequency responses of most pigeon SCPAs. 相似文献
55.
Prediction of postoperative morbidity after lung resection using an artificial neural network ensemble 总被引:4,自引:0,他引:4
OBJECTIVE: To propose an ensemble model of artificial neural networks (ANNs) to predict cardio-respiratory morbidity after pulmonary resection for non-small cell lung cancer (NSCLC). METHODS: Prospective clinical study was based on 489 NSCLC operated cases. An artificial neural network ensemble was developed using a training set of 348 patients undergoing lung resection between 1994 and 1999. Predictive variables used were: sex of the patient, age, body mass index, ischemic heart disease, cardiac arrhythmia, diabetes mellitus, induction chemotherapy, extent of resection, chest wall resection, perioperative blood transfusion, tumour staging, forced expiratory volume in 1s percent (FEV(1)%), and predicted postoperative FEV(1)% (ppoFEV(1)%). The analysed outcome was the occurrence of postoperative cardio-respiratory complications prospectively recorded and codified. The artificial neural network ensemble consists of 100 backpropagation networks combined via a simple averaging method. The probabilities of complication calculated by ensemble model were obtained to the actual occurrence of complications in 141 cases operated on between January 2000 and December 2001 and a receiver operating characteristic (ROC) curve for this method was constructed. RESULTS: The prevalence of cardio-respiratory morbidity was 0.25 in the training and 0.30 in the validation series. The accuracy for morbidity prediction (area under the ROC curve) was 0.98 by the ensemble model. CONCLUSION: In this series an artificial neural network ensemble offered a high performance to predict postoperative cardio-respiratory morbidity. 相似文献
56.
Gregory M. Acland Susan H. Blanton Bennett Hershfield Gustavo D. Aguirre 《American journal of medical genetics. Part A》1994,52(1):27-33
Breeding studies are reported of a previously undescribed hereditary retinal degeneration identified in the Siberian Husky breed of dog. This disorder clinically resembles the previously reported autosomal recessive canine hereditary retinal degenerations collectively termed progressive retinal atrophy (PRA). However, the pedigree of the propositus, a male Siberian Husky, exhibited an X-linked pattern of transmission. This dog was outcrossed to three phenotypically normal female laboratory Beagles and two of their F1 daughters were bred to a phenotypically normal male Beagle, producing affected males in the F2 generation. Subsequent inbreedings produced further affected males and affected females as well. X-linked transmission was established by exclusion of alternative modes of inheritance and, consequently, the disease has been termed X-linked progressive retinal atrophy (XLPRA). This is the first reported X-linked retinal degeneration in an animal. Because of the many similarities of PRA in dogs to retinitis pigmentosa (RP) in humans, this new disease may not only represent the first animal model of X-linked RP (XLRP) but may well be a true homolog of one of the XLRP loci (RP2, RP3, RP6). It is the first retinal degeneration in dogs that can be assigned to an identified canine chromosome, and the first for which linkage mapping offers a realistic approach to proceed by positional cloning towards identifying the responsible gene locus. © 1994 Wiley-Liss, Inc. 相似文献
57.
José Antonio López-Moreno María Scherma Fernando Rodríguez de Fonseca Gustavo González-Cuevas Walter Fratta Miguel Navarro 《Neuroscience letters》2008
Alcohol, nicotine, and cannabinoid acutely increase the activity of the mesolimbic dopamine (DA) pathway. Although polysubstance consumption is a common pattern of abuse in humans, little is known about dopamine release following pre-exposure to these drugs. The purpose of this study was to test whether alcohol-induced dopamine release into the nucleus accumbens (NAc) shell is modified by different pre-treatments: water (i.g.), alcohol (1 g/kg, i.g.), nicotine (0.4 mg/kg, s.c.), and WIN 55,212-2 (1 mg/kg, s.c.). Male Wistar rats were treated (i.g.) for 14 days with either water or alcohol. In the following 5 days rats were injected (s.c.) with vehicle, nicotine, or WIN 55,212-2. Finally, a cannula was surgically implanted into the NAc shell and alcohol-induced extracellular dopamine release was monitored in freely moving rats. Alcohol (1 g/kg; i.g.) only increased the release of dopamine when animals were previously treated with water. This DA increase was markedly inhibited by (subchronic) treatment (5 days) with nicotine or WIN 55-212-2 as well as by previous (chronic) exposure to alcohol (14 days). These data demonstrate that pre-treatment with nicotine and the cannabinoid agonist WIN 55,212-2 is able to change the sensitivity of the NAc shell in response to a moderate dose of alcohol. Therefore, cannabinoid and nicotine exposure may have important implications on the rewarding effects of alcohol, because these drugs lead to long-lasting changes in accumbal dopamine transmission. 相似文献
58.
Schewe C Goldmann T Grosser M Zink A Schlüns K Pahl S Ulrichs T Kaufmann SH Nerlich A Baretton GB Dietel M Vollmer E Petersen I 《Virchows Archiv : an international journal of pathology》2005,447(3):573-585
The present study is based on the initiative for quality assurance in pathology of the German Society of Pathology and the Professional Association of German Pathologists. Four panel laboratories with experience and expertise in polymerase chain reaction (PCR) detection of Mycobacterium tuberculosis were selected to establish the prerequisites for continuous external laboratory trials, in particular, by providing pre-tested specimens and evaluation criteria for participating institutes. In the first step, the four panel laboratories performed an internal trial to test their own reliability and reproducibility. Paraffin sections and DNA preparations from 34 tissues (25 clinical specimens and 9 controls) totalling to 66 samples were evaluated by each panel institute according to their own protocols. The methodologies differed and are described in detail. Despite these differences, a high degree of inter-laboratory reliability was achieved. In this report, we summarise our results including the correlation with the histology and provide recommendations for applying PCR-based methodology for the detection of mycobacterial DNA in surgical specimens. Supplementary data are available online at (rubric Forschung). Pre-tested specimens are now available for the external trial and can be ordered from the steering institute via Oligene (). All molecular pathology laboratories are invited to participate in this quality assurance initiative. 相似文献
59.
Regulation of hepatocyte activator inhibitor-1 expression by androgen and oncogenic transformation in the prostate
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Knudsen BS Lucas JM Fazli L Hawley S Falcon S Coleman IM Martin DB Xu C True LD Gleave ME Nelson PS Ayala GE 《The American journal of pathology》2005,167(1):255-266
Hepatocyte activator inhibitor-1 (HAI-1) is a transmembrane serine protease inhibitor that regulates the conversion of latent to active hepatocyte growth factor (HGF). Studies supporting a role for the HGF pathway in prostate carcinogenesis prompted an analysis of HAI-1 expression in the prostate. Here we analyze the regulation of HAI-1 expression by androgen, oncogenic transformation, and cancer progression. Immunohistochemical analysis revealed that HAI-1 expression was restricted to prostate epithelium, where staining occurred primarily in basal and atrophic luminal epithelial cells. Compared to normal glands, HAI-1 expression was significantly increased in localized prostate cancer and was present in most prostate cancer metastases. HAI-1 protein expression levels were sensitive to androgen in normal epithelium but not in cancer. Although androgen did not increase HAI-1 protein expression levels in LNCaP cells, it decreased HAI-1 surface expression, consistent with previous data from our group (Martin DB, Gifford DR, Wright ME, Keller A, Yi E, Goodlett DR, Aebersold R, Nelson PS: Quantitative proteomic analysis of proteins released by neoplastic prostate epithelium. Cancer Res 2004, 64:347-355). HAI-1 overexpression in cancer was predictive of prostate-specific antigen recurrence (relative risk, 1.24). These results suggest that HAI-1 regulates the HGF Met axis on prostate epithelial cells and influences HGF mediated tumor invasion and metastasis. 相似文献
60.
Luque-Reca Octavio Soriano-Maldonado Alberto Gavilán-Carrera Blanca Acosta-Manzano Pedro Ariza-Vega Patrocinio del Paso Gustavo A. Reyes Álvarez-Gallardo Inmaculada C. Estévez-López Fernando 《Quality of life research》2022,31(7):2059-2059
Quality of Life Research - 相似文献