RT-PCR-RFLP for genetic diversity analysis of Tunisian Grapevine fanleaf virus isolates in their natural host plants

Genetic diversity was characterized in 20 isolates of Grapevine fanleaf virus (GFLV) recovered from naturally infected grapevine plants (Vitis vinifera) in the North of Tunisia. Viral RNAs were isolated by oligoprobe capture, and a 605 bp fragment containing a part of the viral coat protein gene was amplified by RT-PCR. Sequence variation among isolates was characterized by restriction fragment length polymorphism (RFLP) analysis and confirmed by sequencing. The GFLV infections are found as a complex mixture of closely related genomes. In further studies, RFLP analyses of virus isolates using AluI showed that GFLV populations in Tunisian vineyards consist of two restrictotypes corresponding to distinct sub-populations Sp1 and Sp2. The relative field distribution of these sub-populations showed that Sp2 was more abundant. Individual genomes were recovered by cloning the RT-PCR products. The sequences were found to vary from each other by as much as 11%. Cloning from mixed infections showed that Sp2 are also predominant.     

Pancreastatin, a chromogranin A-derived peptide, inhibits DNA and protein synthesis by producing nitric oxide in HTC rat hepatoma cells

BACKGROUND/AIMS: Pancreastatin, a chromogranin A-derived peptide, has a counter-regulatory effect on insulin action. We have previously characterized pancreastatin receptor and signalling in rat liver and HTC hepatoma cells. A G alpha(q/11)-PLC-beta pathway leads to an increase in [Ca2+]i, PKC and mitogen activated protein kinase (MAPK) activation. These data suggested that pancreastatin might have a role in growth and proliferation, similar to other calcium-mobilizing hormones. METHODS: DNA and protein synthesis were measured as [3H]-thymidine and [3H]-leucine incorporation. Nitric oxide (NO) was determined by the Griess method and cGMP production was quantified by enzyme-linked immunoassay. RESULTS: Contrary to the expected results, we have found that pancreastatin inhibits protein and DNA synthesis in HTC hepatoma cells. On the other hand, when the activity of NO synthase was inhibited by N-monomethyl-L-arginine (NMLA), the inhibitory effect of pancreastatin on DNA and protein synthesis was not only reverted, but a dose-dependent stimulatory effect was observed, probably due to MAPK activation, since it was prevented by PD98059. These data strongly suggested the role of NO in the inhibitory effect of pancreastatin on protein and DNA synthesis, which is overcoming the effect on MAPK activation. Moreover, pancreastatin dose-dependently increased NO production in parallel to cyclic guanosine monophosphate (cGMP). Both effects were prevented by NMLA. Finally, an indirect effect of pancreastatin through the induction of apoptosis was ruled out. CONCLUSIONS: Therefore, the NO and the cGMP produced by the NO-activated guanylate cyclase may mediate the dose-dependent inhibitory effect of pancreastatin on growth and proliferation in HTC hepatoma cells.     

Relationship between intra-household food distribution and coexistence of dual forms of malnutrition

BACKGROUND/OBJECTIVES

The relationship between food intake and nutritional status has been clearly established. Yet, there are only limited studies on food intake among family members and their nutritional status. The study examined the relationship between intra-household food distribution and coexistence of dual forms of malnutrition (DFM) in the same household.

SUBJECTS/METHODS

Households with a malnourished child and overweight mother were categorized as DFM. Intra-household food distribution among family members was reported using ratios, which are a measure of individual intakes as compared to all household member intakes adjusted to RDA.

RESULTS

A 1,899 families were included in the study. The prevalence of DFM was 29.8% (95%CI 26.5-31.2). Children consumed lower amounts of energy (OR 1.34; 95%CI 1.06-1.69, P = 0.011), carbohydrates (OR 1.2; 95%CI1.03-1.61, P = 0.022), protein (OR 1.3; 95%CI 1.03-1.64, P = 0.026), and fat (OR 1.3; 95%CI 1.05-1.66, P = 0.016) than their mothers and other family members. In contrast, mothers consumed more carbohydrates than children and other family members (OR1.24; 95%CI 1.02-1.51, P = 0.03).

CONCLUSIONS

This study is the first to report on the food distribution among family members and its relationship with occurrence of DFM in Indonesia. The results confirm the occurrence of an unequal food distribution between children and mothers, which increases risk of DFM in the household. The results also demonstrate that nutritional education at the household level is important to increase awareness of the impact of DFM.     

Assessment of coronary bypass graft patency by electron-beam computed tomography

Twenty-one patients undergoing coronary artery bypass grafting were prospectively evaluated by conventional selective coronary angiography and electron-beam computed tomography. Eighty bypass grafts (60 saphenous vein and 20 left or right internal mammary artery) were evaluated for patency. The sensitivity and specificity of electron-beam computed tomography were 72% and 100%, respectively; positive and negative predictive values were 100% and 92.5%, respectively. Sensitivity and specificity according to coronary region were: left anterior descending artery, 33% and 100%; diagonal artery, 67% and 100%; circumflex artery, 75% and 100%; right coronary artery, 100% and 100%. Electron-beam computed tomography is relatively accurate and a promising tool for noninvasive evaluation of graft patency after coronary artery bypass graft surgery.     

Low Postprandial Circulating Inactive Ghrelin: Role of Early Satiety in Undernourished Children

Objective

To determine difference in the levels of circulating inactive ghrelin between undernourished and healthy children.

Methods

The present cross-sectional study was conducted in undernourished children from southwestern Iran, from July 2011 through July 2012. Postprandial inactive ghrelin levels were measured in 40 undernourished children and sex- and age-matched healthy controls by enzyme immunoassay.

Results

The levels of postprandial inactive ghrelin were considerably lower in undernourished compared to the healthy children (6.4 vs. 12.9, P?vs. 7.1, P?=?0.032).

Conclusions

Thus, the levels of inactive ghrelin was found to be low in undernourished children compared to healthy children. Early loss of appetite might be a result of low circulating inactive ghrelin levels in the postprandial state in undernourished children.