Sarcoglycanopathies: a multiplex molecular analysis for the most common mutations.

Sarcoglycanopathies (SGpathies) are highly frequent among severely affected limb-girdle muscular dystrophy patients. On the basis of the findings of 5 common mutations in the 4 sarcoglycan (SG) genes in the Brazilian population, we standardized a multiplex polymerase chain reaction-single-strand conformation polymorphism methodology for their concomitant analysis in DNA samples. The test was able to confirm the diagnosis in about 63% of new patients with a suspected SGpathy and was particularly important in patients in advanced stages of the disease, when obtaining a muscle biopsy for analysis may be very difficult. As common mutations have been described in several countries, this multiplex analysis could be useful for the diagnosis of SGpathies if established according to the most prevalent mutations in each population. Besides, even though the disorder studied is rare, the technique could be broadly applicable to other genes and disorders.     

The Testes in Cadmium Intoxication: Morphological and vascular aspects

Cadmium toxicity was tested on young male Wistar rats by injecting 1 mg/ml of cadmium chloride (CdCl2) intra-peritoneally. Post-mortem examination was done 4 hours, 24 hours and 14 days after cadmium administration to observe time-sequence cadmium-induced alterations in vascular permeability of the mesothelium in the pleura, peritoneum and tunica vaginals. This paper mainly reports the alterations observed in the testes. Vascular permeability was assessed by the colloidal carbon technique. Vascular labelling was evidenced as early as 4 hours after CdCl2, injection; 24 hours later severe oedema with leakage of particles to the interstitium and also into the tubules was patent. Fourteen days after CdCl2 administration atrophy of the testes with necrosis of the tubules, fibrosis of the interstitium and vascular thrombosis was found, compatible with chemical castration.     

Determination of the cut-off value of serum alanine aminotransferase in patients undergoing hemodialysis, to identify biochemical activity in patients with hepatitis C viremia.

BACKGROUND: Elevated liver enzymes are infrequent in patients with hepatitis C virus (HCV) infection undergoing chronic hemodialysis (HD), suggesting that the alanine aminotransferase (ALT) is a poor predictor of hepatocellular damage in this population. OBJECTIVE: To establish a more appropriate cut-off value of ALT to identify biochemical activity due to HCV infection in HD patients. STUDY DESIGN: A total of 217 patients, with an average age of 51.2 years, were evaluated between January and October 2002; 130 were males (60%). Serum ALT was measured by a kinetic method in five consecutive monthly blood samples, from which an average was obtained and divided by the upper limit of normal (ULN). HCV antibodies were determined using an enzyme immunoassay, the serum HCV-RNA by nested-PCR and HCV genotype by hybridization of the amplified sequence from the 5'-non-coding region. The cut-off value of ALT was obtained from a ROC curve. RESULTS: Within the 217 patients, 18 (8.3%) were anti-HCV-positive, 17 (7.8%) of whom were also HCV-RNA-positive. Genotype distribution was: 1a=47%; 1b=18%; 3a=35%. Mean ALT/ULN (0.77+/-0.57) of the 18 anti-HCV-positive cases was higher (p<0.001) than the negative group (0.38+/-0.23). The mean ALT/ULN (0.81+/-0.57) of the 17 HCV-RNA-positive cases was also higher (p<0.0001) than the negative cases (0.37+/-0.23). The cut-off value of ALT to distinguish the anti-HCV-positive from negative patients was 0.50% or 50% of the ULN (sensitivity=67%; specificity=83%). According to the HCV-RNA, the cut-off value of ALT was 0.45% or 45% of the ULN (sensitivity=71%; specificity=80%). CONCLUSION: Reducing the cut-off of ALT by half, enables a better identification of biochemical activity in patients with HCV infection on chronic HD.     

Characterization of human papillomavirus infection,P53 and Ki-67 expression in cervix cancer of Mozambican women

In this study, we aimed at evaluating the distribution of HPV types and the expression of P53 and Ki-67 in cervix carcinomas of Mozambican women. Fourty-seven invasive carcinomas, 10 CIN III, and 10 normal cervix were studied. P53 and Ki-67 expression was examined immunohistochemically. HPV infection and HPV types were detected by PCR (GP5+/bio-GP6+) and enzyme-immunoassay, respectively. Expression of P53 and Ki-67 and detection of HPV were as follows: normal cervix--0%, 10%, and 0%, respectively; CIN III--10%, 0%, and 100%, respectively; invasive carcinomas--50%, 55.5%, and 70%, respectively. HPV 16 was identified in 54% of invasive carcinomas, HPV 31, 33, 35, and 45 in 23%, "unidentified" HPV in 19%, and HPV 18 in 4% of invasive carcinomas. No significant associations were observed between P53 expression, Ki-67 expression, and HPV infection. In conclusion, we observed a high frequency of HPV infection in CIN III lesions and invasive carcinomas from Mozambican women, with HPV 16 representing the most frequent viral type. HPV status was not related to P53 and Ki-67 expression. Both P53 and Ki-67 are associated with invasive cervix carcinomas, mainly of the squamous keratinizing histotype.     

Incidence of hospital‐acquired pressure injury: A cohort study of adults admitted to public and private hospitals in Sao Paulo,Brazil

This study aimed to estimate the incidence of hospital‐acquired pressure injury (PI) and its risk factors in inpatient and intensive care units of five hospitals (two public and three private) in the city of Sao Paulo, Brazil. A 6‐month follow‐up prospective cohort study (n = 1937) was conducted from April to September 2013. Baseline and follow‐up measurements included demographic and care information, as well as risk assessments for both undernutrition (NRS‐2002) and PI (Braden scale). Poisson regression with robust variance was used for data analysis. A total of 633 patients (32.60%) showed risk for PI. The incidence rate of PI was of 5.9% (9.9% in public hospitals vs 4.1% in private hospitals) and was higher in intensive care units, compared to inpatient care units (10% vs 5.7%, respectively). Risk for PI increased with age (RR = 1.05; 95% CI 1.04‐1.07); was higher in in public hospitals, compared to private hospitals (RR = 4.39; 95% CI 2.92‐6.61); in patients admitted for non‐surgical reasons compared to those admitted for surgical reasons (RR = 1.91; 95% CI 1.12‐3.27); in patients with longer hospital stays (RR = 1.04; 95% CI 1.03‐1.06); high blood pressure (RR = 1.76; 95% CI 1.17‐2.64); or had a risk for undernutrition (RR = 3.51; 95% CI 1.71‐7.24). Higher scores in the Braden scale was associated with a decreased risk of PI (RR = 0.79; 95% CI 0.75‐0.83). The results of our study indicate that 5.9% of all patients developed PI and that the most important factors that nurses should consider are: patient age, care setting, length of hospitalization, comorbidities, reason for admission and nutrition when planning and implementing PI‐preventative actions.     

Revisiting colorectal cancer animal model – An improved metastatic model for distal rectosigmoid colon carcinoma

Colorectal cancer (CRC) is the second most frequent and fatal cancer in Western countries. Understanding its biology with different incidence along the colon and rectum, genetic profile and how these factors contribute to local/distant progression, has been hampered by the lack of a suitable CRC model.We report a reproducible model, using human CRC cell lines (CL) (WiDr, LS1034, C2BBe1) injected (1?×?10⁷ cells/animal) in RNU rats (n?=?55) which underwent cecostomy and descending colostomy with mucosal-cutaneous fistula of the sigmoid colon. CL were characterized by immunohistochemistry: CK20, CDX2, P53, vimentin, Ki67, CD44, CD133, E-cadherin, β-catenin and CEA; cancer stem cells-immune system interaction was studied and tumor progression was assessed with nuclear medicine imaging (^99mTc-MIBI).Animals developed locally invasive tumors and with WiDr neural invasion was registered. Cancer stem cells were detected in WiDr (CD44 positive). All the cell lines stimulated the immune system, being WiDr the most aggressive. Imaging studies demonstrated tumor uptake.With this CRC model we can study the microenvironment role and tumor-stroma interactions. All CL developed primary disease, but only the WiDR established neural invasion which may represent a metastatic pathway. This model can help unveiling the underlying metastatic mechanisms, and ultimately test better therapeutic approaches for CRC.