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.     

A new technique for double lung transplantation. "The bilateral single lung transplantation". Apropos of ten operations]

The technique of enbloc double lung transplantation described by the Toronto team, with cardiopulmonary bypass, cardioplegia, and tracheal suture is complicated by problems of tracheal anastomosis for its authors. Single lung transplantation has a more straight forward postoperative course and bronchial complications are moderate. We present a new technique of lung transplantation, the "bilateral single lung transplantation". It consists, via a single anterior horizontal incision, of successively performing two single lung transplantations without bypass. This operation has been performed ten times since November 1989. As expected, bronchial healing has been excellent.     

Consumption of Vegetables Is Associated with Systemic Inflammation in Older Adults

It is hypothesized that healthy diets rich in fruits and vegetables (FV) can modulate the inflammatory status in older adults. However, to determine the actual impact of FV on inflammatory status, adiposity level and objectively assessed physical activity (PA) behaviors need to be considered. The aim of the present study was to explore associations between FV intake and biomarkers of systemic inflammation in older adults. Based on a sample of 233 older adults (65–70 years old), the following inflammatory biomarkers were assessed: C-reactive protein (CRP), fibrinogen, interleukin-6 (IL-6), IL-10, IL-18, and monocyte chemoattractant protein-1 (MCP-1). FV intake was assessed by self-report, and PA behaviors encompassing time spent sedentary and in moderate-to-vigorous PA (MVPA) were determined using accelerometers. Associations between FV intake and inflammatory biomarkers were analyzed using stepwise linear regression models while adjusting for several covariates, including health-related food groups, adherence to the MVPA guidelines, total sedentary time, and waist circumference. While no significant associations were observed for the total FV intake, the vegetable intake was inversely associated with levels of IL6 (β = −0.15; p < 0.05). In contrast, fruit intake was not associated with any inflammatory biomarker. In conclusion, our findings indicate beneficial associations between vegetable intake and levels of a pro-inflammatory biomarker in older adults, which strengthens public health efforts to promote vegetable-rich diets in older adults to mitigate age-related systemic inflammation.     

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.     

LC-ESI-MS/MS reveals the formation of reactive intermediates in brigatinib metabolism: elucidation of bioactivation pathways

Brigatinib (BGB) is a newly approved anaplastic lymphoma kinase (ALK) inhibitor. On April 28, 2017, BGB was approved by the U.S. FDA for the treatment of metastatic anaplastic lymphoma kinase-positive non-small cell lung cancer. The toxicity profile of BGB includes nausea, fatigue, diarrhea, elevated lipase, dyspnoea, hypertension, hypoxia, pneumonia, elevated amylase, pulmonary embolism, elevated ALT, hyponatraemia and hypophosphatemia. Using LC-MS/MS, we investigated the in vitro phase I metabolism of for BGB in rat liver microsomes (RLMs). In the in vitro metabolism of BGB, iminium reactive intermediates were trapped by potassium cyanide forming a stable complex that can be characterized by LC-MS/MS. Four BGB in vitro phase I metabolites were identified. In vitro phase I metabolic pathways were N-dealkylation, α hydroxylation and α oxidation. Additionally, three iminium reactive metabolites were found and the bioactivation mechanisms were proposed. A piperidine ring was found to be responsible for BGB bioactivation. The presence of these three reactive metabolites may be the main reason for BGB side effects. A literature review showed no previous article reported the in vitro phase I metabolism study of BGB or structural identification of the formed reactive metabolites.

Four phase I BGB metabolites and three cyano adducts for BGB were detected using LC-MS/MS. The piperidine ring was found to be responsible for BGB bioactivation and the bioactivation pathways are proposed.     

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.     

Primary dural lymphoma: A novel concept of heterogeneous disease

Spinal primary dural lymphoma (PDL) is uncommon with a total of 37 previous well‐documented cases reported, including one diagnosed in the authors' institution. More recently we encountered an additional case of spinal PDL that, similarly to our previous case, was grade 1–2 follicular B‐cell PDL. Our two cases were diagnosed over a 3‐year interval in a 72‐year‐old female and a 74‐year‐old male, respectively. An exhaustive literature review on PDL was performed consequently to reveal that: (i) spinal and cerebral sites of involvement by PDL are constantly mutually exclusive; and (ii) unlike cerebral PDL, which is usually of marginal zone B‐cell type, only two of the 38 cases of spinal PDL were diagnosed as such, diffuse large B‐cell lymphoma being the most commonly encountered type in the spine. This divergence infers that, in contrast to the prevailing concept that PDL is a unique disease group, PDL appears to be rather heterogeneous with a difference in predilection of lymphoma type for the anatomical site of dural involvement. Such a site‐specific lymphoma‐type predilection phenomenon, well‐recognized in other organ systems, has not been acknowledged in PDL. This report brings new insights into PDL, and may contribute to a better understanding of nervous system pathophysiology and lymphoma classification.