ABSTRACT
Cryptococcosis is the second most common invasive fungal infection reported in renal transplant recipients. Tissue granulomatous inflammation is necessary to contain Cryptococcus infection. This study aims to analyze the granuloma patterns and in situ expression of regulatory T (Treg) immune response in tissue samples from 12 renal transplant recipients with cryptococcosis. Fungal isolates were molecularly identified as Cryptococcus neoformans species complex. A detailed characterization of granulomas in tissue samples from 12 kidney transplant recipients with cryptococcosis was described by checking six lung and six skin biopsies by conventional histology and for immunohistochemical detection of CD4 and Treg markers: forkhead box P3 (FoxP3), interleukin (IL)-10 and transforming-growth factor (TGF)-ß. Granulomas were classified as compact, loose or mixed. Patients with mixed (n = 4) and compact (n = 3) granulomatous inflammation patterns were associated with a better prognosis and presented a higher number of CD4+FoxP3+T cells compared to the group of patients with loose granulomas. In counterpart, three out of five patients with loose granulomas died with cryptococcosis. We suggest that Treg may have a protective role in the tissue response to Cryptococcus infection given its association with compact and mixed granulomas in patients with better clinical outcomes.
Subject(s)
Cryptococcosis/etiology , Cryptococcosis/mortality , Kidney Transplantation/adverse effects , Cryptococcosis/diagnosis , Cryptococcosis/epidemiology , Cryptococcus , Cryptococcus neoformans/immunology , Disease Susceptibility/immunology , Humans , Immunocompromised Host , Kidney Transplantation/methods , T-Lymphocytes, Regulatory/immunology , T-Lymphocytes, Regulatory/metabolismABSTRACT
INTRODUCTION: There is scarce data on the clinical profile of adult Brazilian patients with nephrotic syndrome caused by minimal change disease (MCD) and focal segmental glomerulosclerosis. OBJECTIVE: We evaluated the clinical characteristics and response to treatment in adult patients with nephrotic syndrome having a histological diagnosis of MCD or FSGS. METHODS: This is a retrospective analysis of 50 patients with MCD and 120 with FSGS. All patients were initially treated with steroids. The study outcomes were: steroid responsiveness, prevalence of total remission, progression to chronic renal failure and need of renal replacement therapy due to end-stage renal disease (ESRD). RESULTS: Initial serum creatinine level was 24% higher among patients with FSGS (p = 0.02), and proteinuria levels were 36% higher in MCD (p < 0.001). Patients with MCD were sensitive to steroid therapy in 80% of the cases, with total remission in 74%, while patients with FSGS were sensitive in 58% (p = 0.01), with total remission in 30% (p = 0.002). Patients with FSGS had an acute renal failure prevalence of 39% (vs. 12%, p = 0.013) and ESRD of 10% (vs. 0%, p < 0.001). Steroid responsiveness reduced in 83% the risk of ESRD (p < 0.001), while total remission was associated to a reduction in risk of 89% (p < 0.001). CONCLUSION: A positive response to steroid therapy was the most important factor related with preservation of renal function and FSGS was related with less steroid responsiveness.
Subject(s)
Glomerulosclerosis, Focal Segmental/drug therapy , Nephrosis, Lipoid/drug therapy , Nephrotic Syndrome/etiology , Steroids/therapeutic use , Adult , Brazil , Creatinine/blood , Female , Glomerulosclerosis, Focal Segmental/complications , Humans , Longitudinal Studies , Male , Nephrosis, Lipoid/complications , Nephrotic Syndrome/drug therapy , Proteinuria/diagnosis , Retrospective Studies , Young AdultABSTRACT
Resumo Introdução: O perfil clínico de pacientes brasileiros adultos com síndrome nefrótica por doença de lesões mínimas (LM) e glomeruloesclerose segmentar e focal (GESF) é pouco conhecido. Objetivo: Avaliamos as características clínico-laboratoriais e resposta a tratamento em pacientes adultos com síndrome nefrótica e diagnósticos histológicos de LM ou GESF. Métodos: Fez-se a análise retrospectiva de 50 pacientes adultos com LM e 120 com GESF. Todos os pacientes foram inicialmente tratados com corticosteroide. Os desfechos do estudo foram: resposta a corticosteroide, prevalência de remissão total, progressão para doença renal crônica estágio 5 (DRC5) e necessidade de terapia de substituição renal por DRC5. Resultados: Níveis iniciais de creatinina sérica foram 24% mais elevados entre pacientes com GESF (p = 0,02) e os de proteinúria foram 36% mais altos em LM (p < 0,001). Pacientes com LM foram córtico-sensíveis em 80% dos casos, com remissão total em 74%, e os pacientes com GESF em 58% (p = 0,01), com remissão total em 30% (p = 0,002). A prevalência de insuficiência renal aguda em pacientes com GESF foi de 39% (vs. 12%, p = 0,013) e DRC5 de 10% (vs. 0%, p < 0,001). Remissão completa ou parcial com o uso de corticosteroide reduziu em 83% o risco de DRC5 (p < 0,001) e remissão total associou-se a redução no risco de DRC5 de 89% (p < 0,001). Conclusão: A resposta positiva à corticoterapia foi o fator mais importante relacionado à preservação da função renal ao longo de mais de uma década de seguimento, e GESF relacionou-se a menor índice de resposta a corticosteroide.
Abstract Introduction: There is scarce data on the clinical profile of adult Brazilian patients with nephrotic syndrome caused by minimal change disease (MCD) and focal segmental glomerulosclerosis. Objective: We evaluated the clinical characteristics and response to treatment in adult patients with nephrotic syndrome having a histological diagnosis of MCD or FSGS. Methods: This is a retrospective analysis of 50 patients with MCD and 120 with FSGS. All patients were initially treated with steroids. The study outcomes were: steroid responsiveness, prevalence of total remission, progression to chronic renal failure and need of renal replacement therapy due to end-stage renal disease (ESRD). Results: Initial serum creatinine level was 24% higher among patients with FSGS (p = 0.02), and proteinuria levels were 36% higher in MCD (p < 0.001). Patients with MCD were sensitive to steroid therapy in 80% of the cases, with total remission in 74%, while patients with FSGS were sensitive in 58% (p = 0.01), with total remission in 30% (p = 0.002). Patients with FSGS had an acute renal failure prevalence of 39% (vs. 12%, p = 0.013) and ESRD of 10% (vs. 0%, p < 0.001). Steroid responsiveness reduced in 83% the risk of ESRD (p < 0.001), while total remission was associated to a reduction in risk of 89% (p < 0.001). Conclusion: A positive response to steroid therapy was the most important factor related with preservation of renal function and FSGS was related with less steroid responsiveness.
Subject(s)
Humans , Male , Female , Adult , Young Adult , Steroids/therapeutic use , Glomerulosclerosis, Focal Segmental/drug therapy , Nephrosis, Lipoid/drug therapy , Nephrotic Syndrome/etiology , Proteinuria/diagnosis , Brazil , Glomerulosclerosis, Focal Segmental/complications , Retrospective Studies , Longitudinal Studies , Creatinine/blood , Nephrosis, Lipoid/complications , Nephrotic Syndrome/drug therapyABSTRACT
BACKGROUND: The purpose of this study was to understand the role of lymphomononuclear inflammation (nephritis) in the renal allograft medulla of transplant recipients with acute dysfunction, by comparing the immunophenotype of inflammatory cells present in the medulla and cortex of kidney graft biopsies. METHOD: This is a retrospective study of 113 renal allograft needle biopsies, presenting with medullary nephritis, divided into two groups according to the main location of nephritis: in cortical and medullary regions (corticomedullary nephritis) or exclusively in the medullary region (medullary nephritis). We performed immunohistochemistry (IHC) of the cells composing the inflammatory foci, using anti-CD4, CD8, CD20, CD68, and CD138 antibodies, respectively for T-helper cells, cytotoxic T cells, B lymphocytes, macrophages and plasmocytes. The clinical follow-up of the patients was correlated with the morphological findings. RESULTS: The nephritis was corticomedullary in 66 of the 113 cases (58.4%) and exclusively medullary in the remaining 47 cases (41.6%). The immunophenotype of the inflammatory cells was similar in the cortical and medullary compartments and were mainly: cytotoxic T lymphocytes (CD8) and macrophages CD68. The immunosuppressive therapeutic response to acute cellular rejection (ACR), based on decreasing of serum creatinine values, was 81.8% in the patients of the corticomedullary nephritis group and 63.6% in those of the medullary nephritis group. CONCLUSION: Medullary nephritis in renal allograft biopsies may indicate ACR, as could be noted by the immunophenotype, which presented the same cellular mediators of rejection seen in the allograft cortex, and by the positive immunosuppressive therapeutic response observed in most patients.
Subject(s)
Graft Rejection/diagnosis , Kidney Medulla/pathology , Kidney Transplantation , Nephritis/diagnosis , Adult , Allografts , Biopsy, Needle , Female , Graft Rejection/immunology , Humans , Immunohistochemistry , Immunophenotyping , Kidney Medulla/immunology , Male , Nephritis/immunology , Retrospective StudiesABSTRACT
BACKGROUND: Late acute rejection (LAR) has been associated with inferior kidney allograft outcomes. METHODS: We retrospectively evaluated 355 episodes of biopsy-confirmed LAR in a cohort of 5758 kidney transplants performed between 1998 and 2008. Estimated glomerular filtration rate was obtained before, at, and after each LAR episode as well as histology and treatment. Associations of LAR with subsequent death or graft loss were estimated with Cox proportional regression analysis. RESULTS: A total of 215 patients had 1 episode, 57 had 2 episodes, and 13 had 3 episodes of LAR. Rates of LAR-free survival were 97.4% at 1 year and 93.7% at 5 years. Estimated glomerular filtration rate decreased after each episode of LAR (56±21 vs. 44±18 vs. 36±11 mL/min/1.73 m, P<0.01). The majority of rejections were Banff IA or less, but the chronicity scores as well as plasma cell infiltrates increased after each LAR. All patients requiring dialysis lost their grafts. In a multivariable analysis, the severity of histological score (risk ratio [RR], 3.5; 95% confidence interval [CI], 1.58-7.87; P<0.001), the need for dialysis at LAR (RR, 3.31; 95% CI, 1.44-7.59; P<0.001), and treatment with methylprednisolone (RR, 2.31; 95% CI, 1.07-4.94; P=0.03) were independently associated with graft loss at 5 years, whereas tacrolimus and mycophenolate use was associated with reduced risk (RR, 0.46; 95% CI, 0.25-0.87; P<0.001). CONCLUSIONS: The prevalence and recurrence of LAR are considerable and associated with increased incidence of graft loss. Patients who need dialysis during LAR should be carefully evaluated owing to the high prevalence of graft failure.
Subject(s)
Graft Rejection/pathology , Kidney Transplantation/adverse effects , Kidney/pathology , Acute Disease , Adult , Female , Glomerular Filtration Rate , Humans , Male , Middle Aged , Proportional Hazards Models , Retrospective Studies , Treatment OutcomeABSTRACT
The purpose of this study was to evaluate the association of post-transplant soluble CD30 (sCD30) levels, isolated or in combination with of anti-HLA class II antibodies and of serum creatinine levels, with kidney graft loss due to chronic allograft nephropathy (CAN), and type of lesions in graft biopsies for cause. The study comprised 511 first kidney graft recipients, transplanted at a single center, with a graft functioning for at least 2.8 years. A single blood sample was collected from each patient. sCD30 levels were determined by ELISA, and HLA antibodies by Luminex assay. The minimum follow-up after testing was 9.3 years. High sCD30 levels, set at sCD30 ≥ 34.15 ng/mL, the presence of HLA class II antibodies, and serum creatinine ≥ 1.9 mg/dL were independently associated with CAN-graft loss (P values <0.0001, 0.05, <0.0001, respectively), and the combined hazard ratio for CAN-graft loss was 20.2. Analyses of 166 biopsies for cause showed that high sCD30 levels and creatinine were independently associated with interstitial lesions. Post-transplant sCD30 serum levels, especially in conjunction with information regarding HLA class II antibodies and serum creatinine levels, provide valuable information regarding graft outcome and could be useful for the management of kidney transplant recipients.
Subject(s)
Graft Rejection/blood , Isoantibodies/blood , Ki-1 Antigen/blood , Kidney Diseases/blood , Kidney Transplantation , Adolescent , Adult , Aged , Child , Child, Preschool , Creatinine/blood , Creatinine/immunology , Female , Follow-Up Studies , Graft Rejection/immunology , Histocompatibility Antigens Class II/immunology , Histocompatibility Antigens Class II/metabolism , Humans , Isoantibodies/immunology , Ki-1 Antigen/immunology , Kidney Diseases/etiology , Kidney Diseases/immunology , Kidney Diseases/prevention & control , Male , Middle Aged , Retrospective StudiesABSTRACT
The goal of this study was to investigate the expression of some metalloendopeptidases in squamous cell carcinomas of the oropharynx as well as its relation to histological differentiation, staging of disease, and prognosis. Paraffin blocks from 21 primary tumors were obtained from archives of the Department of Pathology, Paulista Medical School, Federal University of Sao Paulo, UNIFESP/EPM. Immunohistochemistry was used to detect the expression of EP24.15 and EP24.16 by means of tissue microarrays. Expression of EP24.15 or EP24.16 was not correlated with the stage of disease, histopathological grading or recurrence in squamous cell carcinomas of the oropharynx. In summary, our results support the notion that EP24.15 and EP24.16 are expressed in carcinoma of the oropharynx; however, these do not appear to be suitable biomarkers for histological grading, disease stage or recurrence as depicted by tissue microarrays and immunohistochemistry.
Subject(s)
Carcinoma, Squamous Cell/enzymology , Carcinoma, Squamous Cell/pathology , Metalloendopeptidases/analysis , Oropharyngeal Neoplasms/enzymology , Oropharyngeal Neoplasms/pathology , Adult , Aged , Biomarkers, Tumor/analysis , Female , Humans , Male , Middle Aged , Neoplasm Grading , Neoplasm Staging , Oropharynx/pathology , Prognosis , Tissue Array AnalysisABSTRACT
We studied p38 phosphorylation and its intracellular localization during p53 and Puma (a p53 upregulated modulator of apoptosis) apoptotic signaling pathway in bone marrow granulocytes in mice irradiated in vivo and the role of the radioprotector amifostine in ameliorating these responses. Sixty-four C57BL mice were randomly assigned in two non-irradiated (Ami-/rad- and Ami+/rad-) and two irradiated (Ami-/rad+ and Ami+/rad+) groups. Animals received 400mg/kg of amifostine i.p. 30 min prior to a single whole body radiation dose of 7Gy. The experiments were performed using immunohistochemistry for caspase-3, cleaved caspase-3, p53, p-p53 (Ser 15), Puma, p38 and p-p38 (Thr 180/Tyr 182) protein expression. In addition transmission electron microscopy was used for ultrastructural characterization of apoptosis. Data showed that: (i) amifostine significantly reduced the number of apoptotic cells, (ii) p-p53 and Puma proteins were strongly immunostained in granulocytes after irradiation (Ami-/rad+), (iii) amifostine decreased the immunostaining of the proteins (Ami+/rad+), (iv) p38 was immunolocalized in physiological conditions in the nucleus and cytoplasm of granulocytes and neither radiation nor amifostine changed the protein immunostaining or its subcellular distribution, but influenced its activation, (v) radiation-induced p38 phosphorylation and its cytoplasmic accumulation during apoptosis signaling in granulocytes after whole body high radiation dose and amifostine markedly reduced these effects.
Subject(s)
Amifostine/pharmacology , Apoptosis/physiology , Granulocytes/drug effects , Granulocytes/metabolism , Radiation-Protective Agents/pharmacology , p38 Mitogen-Activated Protein Kinases/metabolism , Animals , Apoptosis/drug effects , Granulocytes/cytology , Granulocytes/radiation effects , Immunohistochemistry , Male , Mice , Mice, Inbred C57BL , Microscopy, Electron, Transmission , Phosphorylation/drug effects , Protein Transport/drug effects , Signal Transduction/drug effectsABSTRACT
BACKGROUND: There are few reports of glomerulonephritis (GN) with crescents and a rapidly progressive course that lead to a diagnosis of a previously unsuspected B-cell dyscrasia. CASE PRESENTATION: We report a case of rapidly progressive GN: the patient showed no evidence of etiology at the time of biopsy and was diagnosed as IgA multiple myeloma (MM) during investigation based on a renal biopsy. He presented diffuse proliferative and exudative GN and marked plasma cell infiltration of the kidney. CONCLUSION: The present case raises the possibility that proliferative GN with crescents may be a rare mode of presentation of MM.
ABSTRACT
AIM: The aim of this study was to investigate the expressions of cell cycle regulatory proteins such as p53, p16, p21, and Rb in squamous cell carcinoma of the oropharynx and their relation to histological differentiation, staging of disease, and prognosis. PATIENTS AND METHODS: Paraffin blocks from 21 primary tumors were obtained from archives of the Department of Pathology, Paulista Medical School, Federal University of Sao Paulo, UNIFESP/EPM. Immunohistochemistry was used to detect the expression of p53, p16, p21, and Rb by means of tissue microarrays. RESULTS: Expression of p53, p21, p16 and Rb was not correlated with the stage of disease, histopathological grading or recurrence in squamous cell carcinoma of the oropharynx. CONCLUSION: Taken together, our results suggest that p53, p16, p21 and Rb are not reliable biomarkers for prognosis of the tumor severity or recurrence in squamous cell carcinoma of the oropharynx as depicted by tissue microarrays and immunohistochemistry.
Subject(s)
Carcinoma, Squamous Cell/metabolism , Carcinoma, Squamous Cell/pathology , Cell Cycle Proteins/metabolism , Oropharyngeal Neoplasms/metabolism , Oropharyngeal Neoplasms/pathology , Protein Array Analysis , Biomarkers, Tumor/metabolism , Cyclin-Dependent Kinase Inhibitor p16/metabolism , Cyclin-Dependent Kinase Inhibitor p21/metabolism , Humans , Immunohistochemistry , Prognosis , Retinoblastoma Protein/metabolism , Severity of Illness Index , Tumor Suppressor Protein p53/metabolismABSTRACT
AIM: We aimed to evaluate the amount and quality of the RNA obtained from lymph nodes of non-Hodgkin lymphomas (NHLs) patients using fine-needle aspiration cytology (FNAC), and to develop strategies to overcome eventual technical drawbacks. MATERIALS AND METHODS: Twenty-six patients with NHL and 10 tonsils from children submitted to tonsillectomy underwent FNAC. The aspirates were performed using both cytoaspirator (sample A) and syringe and needle (sample B). The RNA was extracted using Trizol reagent and transcribed with the Superscript kit (Invitrogen). The quality of RNA was verified through the amplification of a beta-actin 155-bp fragment. RESULTS: Fifty-two NHL and 20 tonsil samples were analyzed. The total amount of RNA in the tonsil samples varied from <1.0 to 6.2 microg with cytoaspirator (A) and from <1.0 to 4.7 microg with syringe and needle (B). The total amount of RNA obtained from NHL varied from <1.0 to 6.5 microg with cytoaspirator (A) and <1.0 to 5.5 microg with syringe and needle. In an attempt to increase the amounts of RNA in each sample, we standardized the polyAPCR technique, which increased by 10 times the amount of cDNA in most of the test and control samples. The efficiency of the reaction was verified through the amplification of beta-actin, in which 100% of the test and control samples were amplified. When polyAPCR cDNA and nonamplified cDNA samples were paired to be evaluated by real-time PCR, using glyceraldehyde-3-phosphate dehydrogenase as the constitutive gene and nuclear factor-kappa B and NFkappaBIA as target genes, there was equivalence in the amplifications of 100% of the 15 evaluated samples. CONCLUSIONS: Our results showed that FNAC, obtained either by cytoaspirator or syringe and needle, is a good source of small amounts of RNA. The polyAPCR technique significantly increased the amount of genomic material, which might be a cDNA source for future gene expression studies.
Subject(s)
Gene Expression Profiling/methods , Lymph Nodes/pathology , Lymphocytes , Lymphoma, Non-Hodgkin/pathology , Pathology, Molecular/methods , RNA/isolation & purification , Actins/genetics , Biopsy, Fine-Needle , Child , Child, Preschool , DNA, Complementary/genetics , Gene Expression Profiling/standards , Humans , Pathology, Molecular/standards , Polymerase Chain Reaction/methods , RNA/geneticsABSTRACT
Renal transplant patients with stable graft function and proximal tubular dysfunction (PTD) have an increased risk for chronic allograft nephropathy (CAN). In this study, we investigated the histologic pattern associated with PTD and its correlation with graft outcome. Forty-nine transplant patients with stable graft function were submitted to a biopsy. Simultaneously, urinary retinol-binding protein (uRBP) was measured and creatinine clearance was also determined. Banff's score and semi-quantitative histologic analyses were performed to assess tubulointerstitial alterations. Patients were followed for 24.0 + or - 7.8 months. At biopsy time, mean serum creatinine was 1.43 + or - 0.33 mg/dl. Twelve patients (24.5%) had uRBP > or = 1 mg/l, indicating PTD and 67% of biopsies had some degree of tubulointerstitial injury. At the end of the study period, 18 (36.7%) patients had lost renal function. uRBP levels were not associated with morphologic findings of interstitial fibrosis and tubular atrophy (IF/TA), interstitial fibrosis measured by Sirius red or tubulointerstitial damage. However, in multivariate analysis, the only variable associated with the loss of renal function was uRBP level > or = 1 mg/l, determining a risk of 5.290 of loss of renal function (P = 0.003). Renal transplant patients who present PTD have functional alteration, which is not associated with morphologic alteration. This functional alteration is associated to progressive decrease in renal function.
Subject(s)
Kidney Transplantation/methods , Kidney Tubules/pathology , Transplantation, Homologous/methods , Adult , Biopsy , Female , Fibrosis , Graft Survival , Humans , Kidney/metabolism , Male , Middle Aged , Prognosis , Retinol-Binding Proteins, Cellular/metabolism , Treatment OutcomeABSTRACT
This study was undertaken to investigate the immunoexpression of Bcl-2 family proteins (Bcl-2, Bcl-x, Bax, Bak and Bad) and to evaluate the correlation between the immunoexpression of these proteins and that of cleaved caspase 3, Ki-67 and p53. A tissue microarray (TMA) paraffin block was constructed using gastric carcinoma tissue (test group) and adjacent normal gastric mucosa (control group) from 87 patients who had not previously undergone radiotherapy or chemotherapy. Sections from the TMA block (4 µm) were subjected to immunohistochemistry for Bcl-2, Bcl-x, Bax, Bak, Bad, p53, Ki-67 and cleaved caspase-3. The slides were evaluated by the semi-quantitative method, and the scores obtained (intensity vs. percentage of staining) were correlated with one another and with the apoptotic index, cellular proliferation and data regarding patient survival. The studied proteins were present in the tumor tissue and in the normal gastric mucosa, but at different intensities and with differences in the number of positive cells. There was an association between tumor size and p53 expression, and intestinal type adenocarcinoma was positively correlated with the expression of Bax, Bad and Ki-67. The immunoexpression of Bcl-x, Bak, Bad, p53 and Ki-67 showed statistically significant differences between the tumor tissue and the adjacent normal gastric mucosa. There was an association between the expression of Bax, Bak and Bad in the normal gastric mucosa. No correlation between patient survival and the expression of these proteins was observed. Overexpression of the Bcl-x protein in the adenocarcinomas and the difference in Bcl-x expression between the test and the control group may be related to the anti-apoptotic effect of this protein. The reduced expression of Bak and Bad and the increased expression of p53 and Ki-67 in the adenocarcinomas demonstrate the imbalance between apoptosis and cellular proliferation, which results in uncontrolled tumor cell proliferation.
ABSTRACT
B lymphocyte infiltration in renal acute allograft rejection has been associated with steroid resistance and poor outcomes. We aimed to measure CD20 mRNA in urine of renal transplant patients with graft dysfunction and correlate with the histological diagnosis and immunohistochemical (IH) staining for CD20. A total of 48 urine samples were analyzed (21 with acute rejection, 10 with chronic allograft nephropathy, 11 with unspecific tubular lesions, 3 with acute pyelonephritis and 3 with polyomavirus nephropathy). Higher urinary CD20 levels associated with a positive IH staining for CD20 (>50 positive cells/HPF) in renal tissue (p=0.04), with a sensitivity of 83.3% and a specificity of 51.6%. Within the acute rejection group, a positive staining for CD20 was not associated with graft loss, steroid resistance or lack of return to basal creatinine after treatment, but was associated with higher serum creatinine at 3 and 6 months, 1 and 2 years after the acute episode (p<0.05). In conclusion, we showed that urinary levels of CD20 detected by RT-PCR had a high sensitivity for CD20+ staining in the corresponding renal tissue, but with a low specificity. Patients with clusters of CD20+ cells >50/HPF had higher serum creatinine after 2 years of follow up.
Subject(s)
Antigens, CD20/biosynthesis , B-Lymphocytes/immunology , Graft Rejection/urine , Kidney Transplantation/immunology , RNA, Messenger/urine , Acute Disease , Adult , B-Lymphocytes/metabolism , Biomarkers/urine , Chronic Disease , Creatinine/blood , Creatinine/urine , Female , Graft Rejection/pathology , Humans , Kidney Transplantation/pathology , Male , Middle Aged , Sensitivity and Specificity , Transplantation, Homologous/immunology , Transplantation, Homologous/pathologyABSTRACT
Development and standardization of reliable methods for detection of Mycobacterium tuberculosis in clinical samples is an important goal in laboratories throughout the world. In this work, lung and spleen fragments from a patient who died with the diagnosis of miliary tuberculosis were used to evaluate the influence of the type of fixative as well as the fixation and paraffin inclusion protocols on PCR performance in paraffin embedded specimens. Tissue fragments were fixed for four h to 48 h, using either 10% non-buffered or 10% buffered formalin, and embedded in pure paraffin or paraffin mixed with bee wax. Specimens were submitted to PCR for amplification of the human beta-actin gene and separately for amplification of the insertion sequence IS6110, specific from the M. tuberculosis complex. Amplification of the beta-actin gene was positive in all samples. No amplicons were generated by PCR-IS6110 when lung tissue fragments were fixed using 10% non-buffered formalin and were embedded in paraffin containing bee wax. In conclusion, combined inhibitory factors interfere in the detection of M. tuberculosis in stored material. It is important to control these inhibitory factors in order to implement molecular diagnosis in pathology laboratories.
Subject(s)
Lung/microbiology , Mycobacterium tuberculosis/genetics , Polymerase Chain Reaction/methods , Spleen/microbiology , Tuberculosis/diagnosis , Animals , Fixatives , Formaldehyde , Humans , Mycobacterium tuberculosis/isolation & purification , Paraffin Embedding , Tissue Fixation/methodsABSTRACT
Development and standardization of reliable methods for detection of Mycobacterium tuberculosis in clinical samples is an important goal in laboratories throughout the world. In this work, lung and spleen fragments from a patient who died with the diagnosis of miliary tuberculosis were used to evaluate the influence of the type of fixative as well as the fixation and paraffin inclusion protocols on PCR performance in paraffin embedded specimens. Tissue fragments were fixed for four h to 48 h, using either 10 percent non-buffered or 10 percent buffered formalin, and embedded in pure paraffin or paraffin mixed with bee wax. Specimens were submitted to PCR for amplification of the human beta-actin gene and separately for amplification of the insertion sequence IS6110, specific from the M. tuberculosis complex. Amplification of the beta-actin gene was positive in all samples. No amplicons were generated by PCR-IS6110 when lung tissue fragments were fixed using 10 percent non-buffered formalin and were embedded in paraffin containing bee wax. In conclusion, combined inhibitory factors interfere in the detection of M. tuberculosis in stored material. It is important to control these inhibitory factors in order to implement molecular diagnosis in pathology laboratories.
O desenvolvimento e a padronização de métodos confiáveis para a detecção de Mycobacterium tuberculosis em amostras clínicas é um objetivo importante nos laboratórios de todo o mundo. Neste trabalho, fragmentos de pulmão e baço de paciente que morreu com o diagnóstico de tuberculose miliar foram usados para avaliar a influência do tipo de fixador e dos protocolos de fixação e inclusão em parafina na performance da PCR. Fragmentos de tecido foram fixados por quatro h a 48 h, usando formalina não tamponada a 10 por cento ou formalina tamponada a 10 por cento e incluídos em parafina pura ou misturada a cera de abelha. As amostras foram submetidas a PCR para amplificação do gene da beta-actina humana e, separadamente, para amplificação da sequência de inserção IS6110, específica do complexo M. tuberculosis. O resultado da amplificação do gene da beta-actina foi positivo em todas as amostras. Não foram gerados amplicons na PCR-IS6110 em amostras de tecido pulmonar fixadas usando formalina não tamponada a 10 por cento e incluídas em parafina com cera de abelha. Em conclusão, fatores inibitórios combinados interferiram na detecção de M. tuberculosis em material de arquivo. É importante controlar estes fatores inibitórios para poder implementar o diagnóstico molecular em laboratórios de patologia.
Subject(s)
Animals , Humans , Lung/microbiology , Mycobacterium tuberculosis/genetics , Polymerase Chain Reaction/methods , Spleen/microbiology , Tuberculosis/diagnosis , Fixatives , Formaldehyde , Mycobacterium tuberculosis/isolation & purification , Paraffin Embedding , Tissue Fixation/methodsABSTRACT
BACKGROUND: Glomerular diseases are an important cause of end-stage renal disease, especially among young adults. However, clinical and epidemiological surveys involving adolescent populations are scarce. AIM: To determine the pattern of glomerulopathies (GP) in adolescents submitted to renal biopsy. METHODS: A retrospective study of patients' records of the Glomerulopathy Section, UNIFESP (Brazil), was performed RESULTS: Among 72 adolescents (12-18 years) with GP, 15.6 +/- 1.5 years, 58.3% females, the most frequent clinical manifestation was nephrotic syndrome (NS, 71%) and focal segmental glomerulosclerosis (FSGS) was the main histological pattern (24%), followed by minimal change disease (MCD, 19.5%). After comparing the main causes of NS in adolescents with those of adults, we found no statistically significant differences in clinical presentation or outcome. Renal failure-free survival of 1 and 5 years for all GP corresponded to 87.9 and 73.6%, respectively (88.5 and 76.3% for NS). CONCLUSIONS: NS was the main manifestation; FSGS and MCD were the most common histological diagnoses. Our data suggest the GP and particularly the NS pattern in adolescents is similar to that of adults, pointing to the need for an adaptation in diagnostic and treatment protocols for this age group, a pattern which corresponds more closely to that of adults.
Subject(s)
Glomerulonephritis/diagnosis , Glomerulonephritis/therapy , Adolescent , Age Factors , Child , Female , Follow-Up Studies , Glomerulonephritis/mortality , Glomerulonephritis/physiopathology , Humans , Male , Retrospective Studies , Survival Rate/trendsABSTRACT
UNLABELLED: Mycophenolate mofetil (MMF) and sirolimus (SRL) are effective immunosuppressive drugs with distinct safety profile. METHODS: Kidney transplant recipients receiving tacrolimus (TAC)-based immunosuppressive regimen were randomized to receive fixed daily doses of MMF (2 g/d, n = 50) or SRL (one loading dose of 15 mg, 5 mg/d till day 7 and 2 mg/d thereafter, n = 50) without induction therapy. RESULTS: No differences were observed in the incidence of the composite (biopsy-confirmed acute rejection, graft loss or death) end-point (18% vs. 16%, p = 1.000), biopsy confirmed acute rejection (12% vs. 14%, p = 1.000), one-yr patient (94% vs. 98%, p = 0.308), graft (92% vs. 98%, p = 0.168), and death-censored graft survival (98% vs. 100%, p = 0.317) comparing patients receiving MMF or SRL respectively. Patients receiving SRL showed worse safety outcomes, higher mean creatinine (1.6 +/- 0.5 mg/dL vs. 1.4 +/- 0.3 mg/dL, p = 0.007), higher proportion of patients with proteinuria (52.0% vs. 10.7%, p = 0.041), higher mean urinary protein concentrations (0.3 +/- 0.5 g/L vs. 0.1 +/- 0.2 g/L, p = 0.012), higher mean cholesterol concentration (217 mg/dL vs. 190 mg/dL, p = 0.030), and higher proportion of patients prematurely discontinued from randomized therapy (26% vs. 8%, p = 0.031). CONCLUSION: In patients receiving TAC, MMF produced similar efficacy but superior safety profile compared with SRL.