José Antonio Tesser Poloni

Anti-inflammatory effects of red pepper (Capsicum baccatum) on carrageenan- and antigen-induced inflammation.

Inflammation is a pivotal component of a variety of diseases, such as atherosclerosis and tumour progression. Various naturally occurring phytochemicals exhibit anti‐inflammatory activity and are considered to be potential drug candidates against inflammation‐related pathological processes. Capsicum baccatum L. var. pendulum (Willd.) Eshbaugh (Solanaceae) is the most consumed species in Brazil, and its compounds, such as capsaicinoids, have been found to inhibit the inflammatory process. However, the anti‐inflammatory effects of C. baccatum have not been characterized. Thus, this study was designed to evaluate the effects of C. baccatum juice in animal models of acute inflammation induced by carrageenan and immune inflammation induced by methylated bovine serum albumin. Pretreatment (30 min) of rats with pepper juice (0.25–2.0 g kg−1) significantly decreased leucocyte and neutrophil migration, exudate volume and protein and LDH concentration in pleural exudates of a pleurisy model. This juice also inhibited neutrophil migration and reduced the vascular permeability on carrageenan‐induced peritonitis in mice. C. baccatum juice also reduced neutrophil recruitment and exudate levels of pro‐inflammatory cytokines TNF‐α and IL‐1β in mouse inflammatory immune peritonitis. Furthermore, we demonstrated that the main constituent of C. baccatum juice, as extracted with chloroform, is capsaicin. In agreement with this, capsaicin was able to inhibit the neutrophil migration towards the inflammatory focus. To our knowledge, this is the first demonstration of the anti‐inflammatory effect of C. baccatum juice and our data suggest that this effect may be induced by capsaicin. Moreover, the anti‐inflammatory effect induced by red pepper may be by inhibition of pro‐inflammatory cytokine production at the inflammatory site.

Evaluation of different urine protocols and DNA extraction methods for quantitative detection of BK viruria in kidney transplant patients

Most transplant centers screen kidney transplant recipients for BK virus (BKV) infection using molecular techniques for the virus load determination. However, there is no consensus about the pre-analytical methods involved in the viral detection. In this study BK viral load was compared by the means of two urine treatment protocols (pelleted vs. whole urine) and two commercial DNA extraction kits for a quantitative PCR (qPCR) experiment. Ten patients who presented decoy cells in their urine sediment were selected for the study. Viral load was considerable higher (>1.5 log) for pelleted urine, in comparison to whole urine but no significant difference was observed between the extraction kits. PCR inhibition did not occur by using pelleted urine. In order to increase test sensitivity to detect BK viruria, pelleted urine should be the preferred urine compartment for qPCR experiments.

Urinary red blood cells: not only glomerular or nonglomerular.

Two main types of red blood cells, isomorphic and dysmorphic, are found in the urine sediment, indicating nonglomerular and glomerular hematuria, respectively. Occasionally, however, other types of red blood cells such as sickle cells, anisocytes, poikilocytes, elliptocytes and dacryocytes can be seen in the urine sediment of patients with hematuria. This paper describes such cases reported in the literature in which such unusual urinary red blood cells have been found and the experience of the authors on this subject.

Cryptococcus within a urinary cast.

A 13-year-old girl with AIDS was admitted to the hospital with neck pain and vomiting. Routine urinalysis revealed an unusual cast containing two encapsulated budding yeast forms entrapped within the matrix (Figure 1). The urine sediment was stained with China ink and revealed free encapsulated budding yeasts (Figure 2). The morphology of the yeast was consistent with Cryptococcus sp. The fungus was also isolated in the cerebrospinal fluid (CSF) and in liver and bone marrow biopsy, characterizing disseminated cryptococcosis. The latex agglutination test was positive in the serum (titer 1:4096). The patient is currently on treatment for the cryptococcal infection with amphotericin B. The diagnosis of disseminated cryptococcosis is usually made by CSF analysis. Urine is an easy-to-obtain sample that may be useful in a search for Cryptococcus sp. Although reports describing cryptococcuria exist, this is the first report of Cryptococcus sp. being entrapped within a urinary cast. Urine microscopy and culture could provide important clues to the presence of cryptococcal infection. n e p h r o l o g y i m a g e http://www.kidney-international.org

Screening for BK virus nephropathy in kidney transplant recipients:comparison of diagnostic tests

Urine cytology and qPCR in blood and urine are commonly used to screen renal transplant recipients for polyomavirus-associated nephropathy (PVAN). Few studies, however, have directly compared these two diagnostic tests, in terms of their performance to predict PVAN. This was a systematic review in which adult (≥ 18 years old) renal transplant recipients were studied. A structured Pubmed search was used to identify studies comparing urine cytology and/or qPCR in urine and plasma samples for detecting PVAN with renal biopsy as the gold standard for diagnosis. From 707 potential papers, there were only twelve articles that matched the inclusion criteria and were analyzed in detail. Among 1694 renal transplant recipients that were included in the review, there were 115 (6.8%) patients with presumptive PVAN and 57 (3.4%) PVAN confirmed. In this systematic review, the qPCR in plasma had better performance for PVAN compared to urine cytopathology. Resumo A citologia urinária e a reação da cadeia da polimerase em tempo real (qPCR) em amostras de sangue e/ou urina são comumente utilizados para rastrear nefropatia associada ao polyomavirus (PVAN), em pacientes transplantados renais. Entretanto, poucos estudos comparam diretamente esses testes diagnósticos quanto ao desempenho para predizer esta complicação. Aqui realizamos uma revisão sistemática na qual foram estudados pacientes transplantados renais adultos (≥ 18 anos). Uma pesquisa estruturada Pubmed foi utilizada para identificar estudos comparando citologia urinária e/ou qPCR em amostras de urina e plasma para detectar PVAN, utilizando a biópsia renal como padrão-ouro para o diagnóstico. Dentre os 707 artigos em potencial, apenas 12 atendiam aos critérios de inclusão e foram analisados em maior detalhe. Foram incluídos 1694 pacientes transplantados renais, entre os quais 115 (6,8%) classificados com PVAN presuntivo e 57 (3,4%) PVAN confirmado. Nessa revisão sistemática, o qPCR no plasma tive melhor desempenho para PVAN em comparação com citopatologia urinária.

Lipoprotein glomerulopathy: a case report of a rare disease in a brazilian child

Lipoprotein glomerulopathy (LPG) is a rare autosomal recessive glomerulopathy associated with the deposition of lipoprotein thrombi in the capillary lumina due to apoE gene mutations. Abnormal plasma lipoprotein profile and marked increase in serum apoliprotein E (apoE) are characteristic clinical data. The compromised patients can present nephrotic syndrome, hematuria, and progressive renal failure. Herein, the authors present the first described case of LPG in a Brazilian male patient, 11 years, who presented with a steroid-resistant nephrotic syndrome. Renal function was normal. Kidney biopsy showed markedly enlarged glomerulus, with dilated capillary loops and weak eosinophilic lipoprotein thrombi in the capillary lumina. Interstitium, tubules, arteries, and veins showed normal histologic aspect. Genotypic study for the apoE gene showed the presence of the alleles E3 and E4. The diagnosis of LPG was then performed. The patient received lipid-lowering treatment. After 2 years of follow-up, renal function is gradually decreasing, with persisting heavy proteinuria, despite a marked decrease in serum cholesterol and triglycerides levels.

Common digital camera and urinary sediment analysis: a tool to be explored

Sir, The common digital camera is a useful tool that assists urinary sediment analysis. Three years ago, in our laboratory , we started taking pictures of urinary sediment. With the database, we were able to organize a Urinalysis Atlas that is constantly updated with new pictures. This atlas has been used to teach our trainees. The photos, taken with a basic Kodak digital camera, have been useful to exchange information on a large number of structures of urinary sediment within a European Centre, particularly concerning the correct identification of the decoy cells on the urinary sediment. According to the article of Fogazzi [1], we have learned about the possibility of identifying this kind of cell without the use of any special stain. The urinary sediment analysis can provide in a few minutes an important clue about a possible reactivation of the polyomavirus infection in the population of kidney transplant patients. To our knowledge, we are the first laboratory in Brazil to report this important finding without any special staining, and the photos made with the digital camera (Figure 1) were an important tool in the process of identifying the decoy NDT Plus (2011) 455 cells. The use of this cheap method can help people in different countries to obtain quality images and allow the professionals to exchange information and improve the diagnosis. Our experience follows the report of Mutter and Brown [2] showing that even with limited technology, it is possible to obtain images of urinary sediment and improve the knowledge of the laboratory personnel.

Detecção quantitativa de vírus BK em receptores de transplante renal: um estudo prospectivo de validação

Abstract Introduction: BK virus (BKV) infection in renal transplant patients may cause kidney allograft dysfunction and graft loss. Accurate determination of BKV viral load is critical to prevent BKV-associated nephropathy (BKVAN) but the cut-off that best predicts BKVAN remains controversial. Objective: To evaluate the performance of a commercial and an in-house qPCR test for quantitative detection of BK virus in kidney transplant recipients. Methods: This was a prospective study with kidney transplant recipients from two large university hospitals in Brazil. Patients were screened for BKV infection every 3 months in the first year post-transplant with a commercial and an in-house real time polymerase chain reaction (qPCR) test. BKVAN was confirmed based on histopathology. The area under the curve for plasma qPCR was determined from receiver operating characteristic analysis. Results: A total of 200 patients were enrolled. Fifty-eight percent were male, 19.5% had diabetes mellitus, and 82% had the kidney transplanted from a deceased donor. BKV viremia was detected in 32.5% and BKVAN was diagnosed in 8 patients (4%). BKVAN was associated with viremia of 4.1 log copies/mL, using a commercial kit. The cut-off for the in-house assay was 6.1 log copies/mL. The linearity between the commercial kit and the in-house assay was R2=0.83. Conclusion: Our study shows that marked variability occurs in BKV viral load when different qPCR methodologies are used. The in-house qPCR assay proved clinically useful, a cheaper option in comparison to commercial qPCR kits. There is an urgent need to make BKV standards available to the international community.

Bright Field Microscopy to Detect Decoy Cells Due to BK Virus Infection in the Fresh and Unstained Urine Sediment in Kidney Allograft Recipients

BK virus (BKV) may reactivate in kidney allograft recipients ultimately leading to BKV nephropathy and graft loss. Decoy cells (DCs) are one of the early marks of BKV reactivation, and these can be detected in the urine sediment.

An Ontology Design Pattern to Represent False-Results

Observations are an important aspect of our society. Arguably, great part of them is captured by means of sensors. Despite the importance of the matter, the ontology of observations and sensors is not well developed, with few efforts dealing with the fundamental questions about their nature. As a result, an important aspect of sensors is overlooked: sensors may fail, producing false-results (i.e. false-positives and false-negatives). The lack of a proper representation of this aspect prevents us from communicating and reasoning about sensor failures, making it harder to assess the correctness of observations and to treat possible errors. In view of this problem, we propose an ontology design pattern (ODP) to represent false-results of sensors. It covers a special case of sensor that exclusively produces positive or negative results regarding the presence of the type of entity the sensor is designed to perceive. The paper introduces the ODP structure as well as its ontological commitments, bringing an example from the biomedical field. Discussion and further research opportunities of research are posed at the end of the paper.