Targeted transcriptomic analysis of pancreatic adenocarcinoma in EUS-FNA samples by NanoString technology.

Background: Integration of transcriptomic testing into EUS-FNA samples is a growing need for precision oncology in pancreatic ductal adenocarcinoma (PDAC). The NanoString platform is suitable for transcriptome profiling in low yield RNA samples. Methods: Inclusion of patients that underwent EUS-FNA cytological diagnosis of pancreatic ductal adenocarcinoma using 19G and/or 22G needles and subsequent surgical resection. Formalin-fixed, paraffin-embedded (FFPE) cytological and surgical samples underwent RNA extraction and transcriptomic analysis using a custom 52-gene NanoString panel of stromal PDAC features. Cell type abundance was quantified in FFPE specimens and correlated. Results: 18 PDAC patients were included. Mean EUS-FNA passes was 2 + 0.7. All FFPE passed the RNA quality control for genomic analysis. Hierarchical clustering on the global gene expression data showed that genes were differentially expressed between EUS and surgical samples. A more enriched cancer-associated fibroblasts and epithelial-mesenchymal transition transcriptomic profile was observed across surgical specimens whereas immunological biomarkers were more represented in EUS-FNA samples. Cytological examination confirmed a scanty representation of CAF and more immunological cell abundance in cytological samples in comparison to surgical specimens. Conclusion: Targeted transcriptomic NanoString profiling of PDAC samples obtained by EUS-FNA is a feasible approach for pre-surgical molecular analysis although stromal CAF/EMT mRNA biomarkers are underrepresented.

Beyond the limits of oxygen: effects of hypoxia in a hormone-independent prostate cancer cell line.

Prostate cancer (PCa) has a high incidence worldwide. One of the major causes of PCa resistance is intratumoral hypoxia. In solid tumors, hypoxia is strongly associated with malignant progression and resistance to therapy, which is an indicator of poor prognosis. The antiproliferative effect and induced death caused by doxorubicin, epirubicin, cisplatin, and flutamide in a hormone-independent PCa cell line will be evaluated. The hypoxia effect on drug resistance to these drugs, as well as cell proliferation and migration, will be also analyzed. All drugs induced an antiproliferative effect and also cell death in the cell line under study. Hypoxia made the cells more resistant to all drugs. Moreover, our results reveal that long time cell exposure to hypoxia decreases cellular proliferation and migration. Hypoxia can influence cellular resistance, proliferation, and migration. This study shows that hypoxia may be a key factor in the regulation of PCa.

Association between dyslipidemia and anthropometric indicators in adolescents.

The dyslipidemia associated with excess weight is a risk profile global call for cardiovascular disease (CVD). The aim of this study was to investigate the association between dyslipidemias and other risk factors for cardiovascular diseases (CVD) in adolescents, considering sexual maturation. A cross-sectional study was carried out with 432 adolescents from public schools, aged 10-19 years. The correlations between the variables from the lipid profile and the Body Mass Index (BMI), waist circumference (WC), waist-to-height ratio (WHtR), sexual maturation, familial history and maternal education were evaluated using Pearson's correlation coefficient. Low high-density lipoprotein cholesterol (HDL-C) was the most prevalent dyslipidemia (50.5%), regardless of gender. There were significant correlations between triglycerides and BMI (r = 0.30, p<0.01), WC (r = 0.32, p < 0.01) and WHtR (r = 0.33, p < 0.01). The linear model, which took into consideration sexual maturation, age and BMI, explain about 1 to 10.4% of the lipid profile variation. The low HDL-c was the most prevalent dyslipidemia in all adolescents and hypertriglyceridemia was most prevalent in overweight adolescents. Associations between dyslipidemias and anthropometric indicators (BMI and RCA) detected in this study can generate the hypothesis of the risk factors for CVD in adolescents.

Assessment of copper and lipid profile in obese children and adolescents.

The aim of this study was to assess erythrocyte and plasma copper concentrations and correlate them with the lipid profile of overweight and obese children and adolescents. The study was performed with 15 overweight and 30 obese children and adolescents, and the results were compared to the control group (21), aged 6-16 yr. Anthropometric assessment was carried out using body mass index (BMI). Total cholesterol, low-density lipoprotein (LDL) cholesterol, high-density lipoprotein (HDL) cholesterol, and triglyceride serum levels were investigated. Erythrocyte and plasma copper levels were determined by atomic absorption spectrophotometry. Greater alterations in the lipid profile were observed in HDL-cholesterol, LDL-cholesterol, and triglyceride levels, with distinctions according to gender. The plasma copper concentrations in the overweight and obese male groups were significantly higher than those in the control group (p = 0.0006). Negative correlations between plasma copper and total cholesterol (r = -0.54) and LDL cholesterol (r = -0.59) were observed in the obese male group. There was no statistical difference in copper erythrocyte concentrations. The obesity associated to disorders in lipid metabolism predisposes to changes in copper plasma concentrations, but there was no alteration in intracellular reserves, which suggests an important homeostatic control to compensate for plasma oscillations and metabolic alterations of the disease.

Relação entre proteína dietética e calciúria em crianças e adolescentes com nefrolitíase / Relation between diet protein and calciuria in children and adolescents with nephrolitiasis

PURPOSE: The objective was to evaluate the quality of diet and the relationship between protein diet and calciuria in children and adolescents with nephrolithiasis. METHODS: Forty-nine children and adolescents (28 male and 21 female; 10.1 +/- 3.16 years old) with nephrolithiasis were included in study. Diet evaluation was performed over a 3 day period in order to determine nutrient consumption. The analysis of diets were carried out by DietWin Clínico 3.0 software. One 24-hour urine sample was collected for the measurement of calcium. Nutritional status was also assessed by Body Mass Index (BMI). RESULTS: The diet of patients containe insufficient energy and calcium. High levels of protein (69.64 +/- 16.42 gm), mainly animal source (65.81 +/- 11.45%) and low levels of calcium (500.95 +/- 284.64 mg) was observed (95%). Analysis of 24 hour urine samples revealed that 25.0% of the patients presented hypercalciuria. A positive correlation (r = 0.26680) between animal protein intake and calciuria was found, in opposite of vegetable protein and calciuria correlation (r = -0.2675). CONCLUSIONS: Animal protein of the diet has a significant effect in urinary excretion of calcium in patients with nephrolithiasis.

Early diagnosis and treatment of leprosy in intradomiciliary contacts in a high prevalence area: Amazon region.

The importance of dermato-neurological examination of intradomiciliary contacts is well known as an important secondary preventive measure in leprosy control, due to the fact that it allows early diagnosis and treatment. This is an intervention trial in an area of high leprosy prevalence (Manaus/Brazil) where the proportion of contacts examined is low. The aim of the study is to assess whether a simple educational session conducted among patients increases contacts examination and leads to early case detection. The intervention group had examined more contacts (p < 0.05) but, paradoxically, presented fewer new cases than the control group. The authors discuss the probable causes for this unexpected outcome, the advantages of the intervention and other related issues.

Polyacrylamide gel electrophoresis as a tool for the taxonomic identification of snakes from the Elapidae and Viperidae families

Polyacrylamide gel electrophoresis (PAGE) for basic proteins may be a useful toll for the characterization of whole snake venoms and for the taxonomic classification of snakes of the Elapidae and Viperidae families. However, due to the close proximity of PAGE was not able to provide an efficient differentiation. This article reports the electrophoretic analysis of several venoms from the genera Micrurus, Bothrops, Bothriopsis, Crotalus and Lachesis and shows a typical and distinctive electrophoretic profile for each species, with intraspecific and geographic variation. Even in cases in which extreme morphological similarities were present, such as between B. jararacussu and B. pirajai ("Bahia jararacussu"), differentiation could be evidenced by PAGE. This simple and sensitive procedure may be applied to similar cases involving basic toxins.

Expression of active, processed ricin in transgenic tobacco.

The cDNA encoding the plant toxin precursor preproricin was introduced into tobacco via Agrobacterium tumefaciens-mediated gene transfer. Transgenic plants were assayed for type II ribosome-inactivating protein expression and activity. Western blot analysis of soluble leaf extracts using anti-ricin a-chain (RTA) antibodies identified 34- and 32-kDa proteins, which were electrophoretically indistinguishable from castor seed RTA. Analysis with anti-ricin b-chain (RTB) antibodies identified both a 34-kDa protein major band, which co-migrated with castor seed RTB, and a 30-kDa protein minor band. Enzyme-linked immunoassay of the transgenic leaf extracts with anti-RTA and anti-RTB indicated microgram per gram production on a fresh weight basis of soluble extractable recombinant ricin. Sugar binding enzyme-linked immunoassay employing an immobilized glycoprotein, asialofetuin, and anti-RTB antibodies confirmed the characteristic type II ribosome-inactivating protein galactose binding lectin activity of the recombinant ricin. The enzymatic activity of recombinant ricin was characterized for cell-free translation inhibition, as well as for overall cytotoxicity. A 50% inhibitory dose of 3 x 10(-11) M was observed for the immunoreactive leaf extract material using a rabbit reticulocyte translation inhibition assay, while a 50% lethal dose of 1 x 10(-12) M was calculated with human T-lymphotropic virus-1 infected leukemic T-cells.

[Iron supplementation effects on zinc bioavailability in a regional diet of northeastern Brazil]. / Efeito da suplementação com ferro na biodisponibilidade de zinco em uma dieta regional do nordeste do Brasil.

The effects of supplementation with iron on the zinc bioavailability of the regional diet of northeastern Brazil (RDN), were investigated. One assay with Wistar rats, feed on RDN and control diets was carried out. The RDN diets contained 16 mg Zn/kg and levels of 35 mg, 70 mg and 140 mg Fe/kg, respectively. The control diets were prepared according to the standards of the Committee on Laboratory Animal Diets, with levels of protein, iron and zinc identical to those of RDN diets. Index of apparent absorption and zinc retained in the femur of the animals were the parameters utilized to measure zinc bioavailability. The results demonstrated that the supplementation with iron decreased the zinc bioavailability, and the effects were seen to affect diet quality and the Fe:Zn ratio. This fact must be taken into consideration in practices such as the fortying of foodstuffs and the administration of vitamin-mineral supplements to populations with nutritional deficiencies.

Teste cicloergometrico em portadores da sindrome de Wolff-Parkinson-White. / Cycloergometric test in patients with Wolff-Parkinson-White syndrome

Foram submetidos a teste cicloergometrico continuo (protocolo habitual) 30 pacientes (22 do sexo masculino) portadores da sindrome de Wolff-Parkinson_white (WPW). A idade variou de 13 a 70 anos. Nao havia evidencia de outras doencas nem estavam em tratamento com antiarritmicos. Durante o teste, nao foram observados sinais de comprometimento da funcao cardiaca nem arritmias significativas (4 pacientes haviam apresentado previamente taquiarritmias paroxisticas comprovadas). A onda delta permaneceu inalterada em 19 pacientes e nos 3 casos de WPW intermitente, o esforco nao determinou o seu aparecimento. Em 8 pacientes, submetidos a teste maximo, desapareceu a onda delta durante o esforco (mantendo-se o segmento PR curto), com frequencia cardiaca superior a 150 bpm. Neste subgrupo, a onda delta reapareceu apos o esforco (1 a 4 min), ainda com frequencia cardiaca superior a 100 bpm. Foi encontrada em 9 pacientes (30%) resposta isquemica indiscutivel, 1 deles evidenciou desaparecimento da onda delta durante o esforco. Os autores comentam a fisiopatologia das alteracoes eletrocardiograficas encontradas durante o exercicio e o mecanismo determinante de arritmias, concluindo que o teste ergometrico nao parece ser um metodo eficiente para inducao de arritmias e para analise de drogas antiarritmicas em portadores de WPW

A technique for isolating and concentrating microfilariae from peripheral blood by gradient centrifugation.

A method for isolating microfilariae from blood cells by Ficoll-Hypaque centrifugation, followed by concentration by conventional centrifugation, is described. A 30 fold concentration and quantitative recovery of parasites was achieved. This technique should be of value for documenting low levels of parasitaemia, and for obtaining relatively pure suspensions of microfilariae for metabolic studies and antigen analysis.