Genotypic resistance testing in HIV by arrayed primer extension

The analysis of mutations that are associated with the occurrence of drug resistance is important for monitoring the antiretroviral therapy of patients infected with human immunodeficiency virus (HIV). Here, we describe the establishment and successful application of Arrayed Primer Extension (APEX) for genotypic resistance testing in HIV as a rapid and economical alternative to standard sequencing. The assay is based on an array of oligonucleotide primers that are immobilised via their 5′-ends. Upon hybridisation of template DNA, a primer extension reaction is performed in the presence of the four dideoxynucleotides, each labelled with a distinct fluorophore. The inserted label immediately indicates the sequence at the respective position. Any mutation changes the colour pattern. We designed a microarray for the analysis of 26 and 33 codons in the HIV protease and reverse transcriptase, respectively, which are of special interest with respect to drug resistance. The enormous genome variability of HIV represents a big challenge for genotypic resistance tests, which include a hybridisation step, both in terms of specificity and probe numbers. The use of degenerated oligonucleotides resulted in a significant reduction in the number of primers needed. For validation, DNA of 94 and 48 patients that exhibited resistance to inhibitors of HIV protease and reverse transcriptase, respectively, were analysed. The validation included HIV subtype B, prevalent in industrialised countries, as well as non-subtype B samples that are more common elsewhere

THE INFLUENCE OF RAT ENDOGENOUS NITROGEN-EXCRETION ON THE ASSESSMENT OF BEAN PROTEIN-QUALITY

32442543

THE USE OF N-15 LABELING FOR THE ASSESSMENT OF LEGUMINOUS PROTEIN DIGESTIBILITY

This study evaluated the digestibility of leguminous protein labeled with N-15, by using nitrogen balance and quantitation of fecal endogenous nitrogen (FEN), determined by isotopic dilution, in order to correct apparent values. Seeds of common beans, cowpea and common pea labeled with 1.000 atoms% of N-15-excess were used as protein sources in diets for 60 male Wistar rats, during a 6-day assay. The real digestibility values obtained with FEN were 77.6, 84.4, and 86.3% for common beans, cowpea and common pea, respectively. They were higher and statistically different (p < 0.05) than true digestibility values, corrected by non-protein diet. FEN showed a direct, moderate and positive relation with weight of dry matter eaten, initial body weight, weight gain and weight of dry matter of feces, the latter showing the highest correlation, with a coefficient r = 0. 8930 at 1% level.391475

EFFECT OF DIETS CONTAINING DRY BEANS (PHASEOLUS-VULGARIS, L) ON THE RAT EXCRETION OF ENDOGENOUS NITROGEN

116122387239

LESIONS OF INTESTINAL EPITHELIUM BY INGESTION OF BEAN LECTINS IN RATS

35431532

EXTRACTION, PARTIAL CHARACTERIZATION AND NUTRITIONAL ASPECTS OF PROTEINS FROM CARIOCA-80 BEANS (PHASEOLUS-VULGARIS, L)

3718810

Low doses of insulin-like growth factor-I improve nitrogen retention and food efficiency in rats with early cirrhosis

Background/Aims: In order to ascertain whether malnutrition is an early-onset feature of liver cirrhosis and whether the anabolic hormone insulinlike growth factor I (IGF-I) could be useful in the treatment of this complication, we analyzed the nutritional alterations present in rats with early-stage liver cirrhosis and the effects of IGF-I on nutritional parameters in these animals. Methods: After a 24 h fast, a N-15-enriched diet was administered for 5 days to normal control rats and to cirrhotic rats receiving subcutaneous injections of vehicle (Group 1) or IGF-I, 2 mu g . 100 g bw(-1) day(-1), (Group 2) during the 5 experimental days. N-15, a stable N isotope, was measured in biological samples by mass spectrometry. Results: Compared with control rats, Group 1 animals showed significant reductions in N intake and food efficiency (p<0.05, both). In addition, the weight of the gastrocnemius muscle, its total N content and the dietary N content of this muscle were significantly lower in Group 1 than in control animals (p<0.05, all). In rats from Group 2, mean values of N intake, food efficiency, gastrocnemius N content and the amount of dietary N incorporated into this muscle were similar to those in control rats, and (with the exception of gastrocnemius N total content) significantly higher than those in non-treated cirrhotic rats (p<0.05, all). Conclusions: A variety of nutritional disturbances were detected in rats from the early stages of liver cirrhosis. Low doses of IGF-I were found to reverse most of these changes. These results stimulate further studies to determine whether IGF-I might be useful in the correction of the malnutrition present in patients with liver cirrhosis.26119120

Extended lymphadenectomy in patients with pancreatic cancer is debatable

Lymph node staging is one of the most important factors in determining the prognosis after resection of pancreatic ductal adenocarcinoma. Despite ongoing efforts to further refine lymph node staging, the debate on the extent of lymphadenectomy during pancreaticoduodenectomy is still open. The purpose of this review was to summarize the evidence about performing standard lymphadenectomy during curative resection of pancreatic cancer. All four prospective randomized controlled trials published concluded that extended lymphadenectomy does not contribute to better oncologic outcome for patients with adenocarcinoma of the pancreatic head. Indeed, one major drawback of extended lymphadenectomy is the higher risk of persistent postoperative diarrhea. No prospective randomized studies could be found on the role of extended lymphadenectomy in patients with adenocarcinoma of the corpus and tail. Based on current evidence there is no indication that extended lymphadenectomy should be performed routinely during resection of pancreatic cancer