Prevalence of the Metabolic Syndrome in a Bulgarian Female Population Referred for Bone Density Testing

Objective: To investigate the prevalence of the metabolic syndrome in Bulgarian women referred for bone density screening. Research Methods and Procedures: This was a cross‐sectional clinical study. Subjects were 444 consecutive 30‐ to 75‐year‐old Bulgarian women recruited from the outpatients referred for bone density testing (mean age, 52.67 ± 15.19 years; mean BMI, 26.10 ± 5.71 kg/m²). Height (centimeters), weight (kilograms), and blood pressure were measured. BMI and waist‐to‐hip ratio were calculated. Fasting plasma glucose, blood lipids, and immunoreactive insulinemia (Bayer Corp.‐Diagnostics Div., Tarrytown, NY) were determined. Body composition was analyzed by bioimpedance on a leg‐to‐leg analyser (Tanita TBF‐215; Tanita Corporation, Tokyo, Japan). Results: Of all women, 56.76% had a BMI > 25 kg/m², 45.95% had a waist circumference > 88 cm, and 64.64% had a waist‐to‐hip ratio > 0.8; 59.90% had hypertension; 4.05% had fasting plasma glucose > 7.0 mM, and 42.79% had fasting morning immunoreactive insulinemia = 16 UI/liter; 23.65% had hypercholesterolemia; and 26.35% had hypertriglyceridemia. The prevalence of the metabolic syndrome in this sample, as defined by the National Cholesterol and Education Program‐Adult Treatment Panel III, was 34.91%, and by the modified World Health Organization definition was 37.16%. Discussion: We concluded that Bulgarian women 30 to 75 years old referred for bone density testing have a high prevalence of the metabolic syndrome. Therefore, large‐scale prevention programs are needed in this field.     

Neuronal nitric oxide synthase immunopositive neurons in cat claustrum—a light and electron microscopic study

Nitric oxide is a unique neurotransmitter, which participates in many physiological and pathological processes in the organism. Nevertheless there are little data about the neuronal Nitric Oxide Synthase immunoreactive (nNOS-ir) neurons and fibers in the dorsal claustrum (DC) of a cat. In this respect the aims of this study were: (1) to demonstrate nNOS-ir in the neurons and fibers of the DC; (2) to describe their light microscopic morphology and distribution; (3) to investigate and analyze the ultrastructure of the nNOS-ir neurons, fibers and synaptic terminals; (4) to verify whether the nNOS-ir neurons consist a specific subpopulation of claustral neurons; (5) to verify whether the nNOS-ir neurons have a specific pattern of organization throughout the DC. For demonstration of the nNOS-ir the Avidin-Biotin-Peroxidase Complex method was applied. Immunopositive for nNOS neurons and fibers were present in all parts of DC. On the light microscope level nNOS-ir neurons were different in shape and size. According to the latter they were divided into three groups-small (with diameter under 15 mum), medium-sized (with diameter from 16 to 20 mum) and large (with diameter over 21 mum). Some of nNOS-ir neurons were lightly-stained while others were darkly-stained. On the electron microscope level the immunoproduct was observed in neurons, dendrites and terminal boutons. Different types of nNOS-ir neurons differ according to their ultrastructural features. Three types of nNOS-ir synaptic boutons were found. As a conclusion we hope that the present study will contribute to a better understanding of the functioning of the DC in cat and that some of the data presented could be extrapolated to other mammals, including human.     

Human papillomavirus prevalence in lung carcinomas in Bulgaria

A possible association between high‐risk human papillomaviruses (HPV) and lung cancer has been investigated for decades with discrepant results. The aim of this study was to determine the prevalence of HPV16 and 18 in Bulgarian patients with lung cancer. Two hundred and nine biopsy specimens from patients with histologically proven lung cancer and without cancer were analyzed. Each sample was subjected to three parallel PCRs using broad spectrum GP5+/6+ primers and type‐specific (TS) primers for HPV types 16 and 18. Of the 132 lung carcinoma samples, 33 (25%) were positive for HPV16 and/or HPV18 by TS PCR whereas only five (3.8%) samples were HPV positive by consensus PCR. All non‐malignant controls were HPV negative. HPV18 was the more prevalent, being found in 11.4% of samples, followed by HPV16 in 9.1% samples; 4.5% of lesions were positive for both HPV16 and HPV18. HPV16/18 were most prevalent in small cell carcinoma (29.2%) and least prevalent in squamous cell carcinoma (23.3%). HPV was only detected in squamous cell carcinoma and adenosquamous carcinoma by consensus PCR. This study revealed a high HPV16/18 prevalence in lung carcinoma samples from Bulgarian patients when TS PCR was used to detect them. The difference between HPV positivity as detected by consensus and by TS PCR was significant, indicating the importance of methodological issues in explaining the discrepancies between previous studies. HPV18 was more common than HPV16. No association between HPV16/18 status and histopathological diagnosis was identified.

     

The Ty1 transposition assay: a new short-term test for detection of carcinogens

An assay based on induction by carcinogens of Ty1 transposition in Saccharomyces cerevisiae is proposed. A tester strain was developed that contains a marked Ty1 element, which allows following the transposition in the genome as a whole and a mutation, which increases cellular permeability. Hypersensitivity to chemical agents, higher cell wall porosity and transformability with plasmid DNA evidenced an enhanced cellular permeability of the tester cells. The increased permeability resulted in higher sensitivity to carcinogens. The treatment with different laboratory carcinogens induced Ty1 transposition rates in the tester strain by a factor of 10 to 20, compared to the controls. The induction is not stress-generated by the cytotoxicity of carcinogens, since treatment with NaN3 at concentrations killing 50% of the cells did not increase the transposition rate. The increase of Ty1 transposition in tester cells is specific for active carcinogens and a positive response with procarcinogens was obtained only in presence of S9 mix. The Ty1 transposition test responded positively to a number of Ames-test or DEL-test negative carcinogens. The positive response of Ty1 test was statistically significant and verified in kinetics and concentration-dependent experiments. It is concluded that the Ty1 transposition test can be used, in addition to the Ames assay, as a short-term test for detection of carcinogens.     

The best control for the specificity of RNAi

RNA interference (RNAi) is revolutionizing functional genomics. However, there are several reasons to be concerned about the specificity and off-target effects of this technique. A recent paper by Kittler et al. describes a straightforward way to validate RNAi specificity, which exploits the increasing availability of bacterial artificial chromosome (BAC) clone resources. Genetic rescue of the RNAi phenotype by BAC transgenesis is the best control yet described for specificity, and has further implications for reverse genetics.     

Differentiation des hydroxy-5 dimethoxy-6,7 ou 7,8 et des triméthoxy-5,6,7 ou 5,7,8 flavones par spectrometrie de masse

The relative abundance of M and M-15 peaks in the mass spectrum allows the differentiation of 5-hydroxy-6,7-dimethoxy- and 5,7,8-trimethoxyflavones (M > M-15) from 5-hydroxy-7,8-dimethoxy and 5,6,7-trimethoxy-flavones (M < M-15) respectively, thus affording a solution to the 5,6,7- or 5,7,8-orientation problem on microscale.     

Skeletal muscle regeneration involves macrophage-myoblast bonding

Regeneration in adult skeletal muscle relies on the activation, proliferation, and fusion of myogenic precursor cells (MPC), mostly resident satellite cells (SC). However, the regulatory mechanism during this process is still under evaluation, with the final aim to manipulate regeneration when the intrinsic mechanism is corrupted. Furthermore, intercellular connections during skeletal muscle regeneration have not been previously thoroughly documented. Our hypothesis was that a direct and close cellular interaction between SC/MPC and invading myeloid cells is a key step to control regeneration. We tested this hypothesis during different steps of skeletal muscle regeneration: (a) the recruitment of activated SC; (b) the differentiation of MPC; (c) myotubes growth, in a mouse model of crush injury. Samples harvested (3 and 5 days) post-injury were screened by light and confocal microscopy. Ultrastructural analysis was performed by conventional transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM) followed by 3D modeling of electron tomography (ET) data. This revealed a new type of interaction between macrophages and myogenic cells by direct heterocellular surface apposition over large areas and long linear distances. In the analyzed volume, regions spaced below 20 nm, within molecular range, represented 31% of the macrophage membrane surface and more than 27% of the myotube membrane. The constant interaction throughout all stages of myogenesis suggests a potential new type of regulatory mechanism for the myogenic process. Thus, deciphering structural and molecular mechanisms of SC-macrophage interaction following injury might open promising perspectives for improving muscle healing.     

Changes in Heterodera glycines Egg Population Density in Continuous Glycine max over Four Years

Soybean cyst nematode, Heterodera glycines, is found throughout soybean production areas of the United States, but the nematode''s distribution is not uniform within states, counties, and individual fields. The goal of this research was to determine the spatial pattern of H. glycines population density in a field in southeastern Missouri and whether it changed over time in the absence of management practices. Geostatistical methods were used to describe and map the distribution of H. glycines over 4 years in a soybean (Glycine max) field in southeastern Missouri. Semivariograms and kriging, an interpolation method, were used to prepare isoarithmic contour maps and associated error maps. In the field studied, fall H. glycines population density (Pf) was poorly related to density the following spring (Pi). The distribution of peak H. glycines population density within the field changed from year to year, although high densities were often detected in the same general region of the field. The patchiness of H. glycines distribution within a field was verified. Yield was not related to H. glycines egg population density at planting, indicating that unmeasured variables were also reducing yield.     

SORLA/SORL1 Functionally Interacts with SPAK To Control Renal Activation of Na+-K+-Cl? Cotransporter 2

Proper control of NaCl excretion in the kidney is central to bodily functions, yet many mechanisms that regulate reabsorption of sodium and chloride in the kidney remain incompletely understood. Here, we identify an important role played by the intracellular sorting receptor SORLA (sorting protein-related receptor with A-type repeats) in functional activation of renal ion transporters. We demonstrate that SORLA is expressed in epithelial cells of the thick ascending limb (TAL) of Henle''s loop and that lack of receptor expression in this cell type in SORLA-deficient mice results in an inability to properly reabsorb sodium and chloride during osmotic stress. The underlying cellular defect was correlated with an inability of the TAL to phosphorylate Na⁺-K⁺-Cl⁻ cotransporter 2 (NKCC2), the major sodium transporter in the distal nephron. SORLA functionally interacts with Ste-20-related proline-alanine-rich kinase (SPAK), an activator of NKCC2, and receptor deficiency is associated with missorting of SPAK. Our data suggest a novel regulatory pathway whereby intracellular trafficking of SPAK by the sorting receptor SORLA is crucial for proper NKCC2 activation and for maintenance of renal ion balance.Sorting-protein-related receptor with A-type repeats (SORLA; also known as SORL1 or LR11) is a member of the VPS10P domain receptor gene family, a group of sorting receptors structurally related to the yeast vacuolar protein sorting 10 protein (VPS10P) (11, 30). VPS10P is a receptor that transports carboxypeptidase Y into the vacuole (the yeast lysosome) (12). VPS10P domain receptors, including SORLA, have been shown to shuttle between the cell surface, early endosomes, and the trans-Golgi network (TGN) in various cell types (16, 26). Based on this distinct trafficking pattern and on their homology to VPS10P, a role for SORLA and other family members as intracellular sorting receptors has been suggested (reviewed in reference 29).Recently, a role for SORLA in protein trafficking was confirmed in neurons in the brain, where it acts as a receptor for the amyloid precursor protein (APP), the main etiological agent in Alzheimer''s disease. SORLA controls the sorting of APP between the TGN and endosomal compartments, thereby regulating proteolytic breakdown into amyloidogenic and nonamyloidogenic products (1, 17, 24).In addition to the brain, expression of SORLA has also been reported in other tissues, including testis, pancreas, smooth muscle, and kidney (11, 15, 30). In the kidney, expression was mapped to the collecting duct epithelium (23). However, the relevance of the receptor for renal physiology remained enigmatic.In this study, we addressed the physiological role of SORLA in the kidney. Contrary to previous reports, we detected significant expression of SORLA in epithelial cells of the thick ascending limb (TAL) of Henle''s loop, the distal convoluted tubule, and the connecting tubule. Lack of receptor expression in SORLA-deficient mice results in an inability to properly reabsorb sodium and chloride under osmotic stress induced by thirsting. The underlying cellular defect was linked to an inability of the TAL to activate Na⁺-K⁺-Cl⁻ cotransporter 2 (NKCC2), the major sodium transporter in the distal nephron. Interaction of SORLA with Ste-20-related proline-alanine-rich kinase (SPAK), the kinase that activates NKCC2, and missorting of SPAK in SORLA-deficient cells suggest a novel regulatory pathway in which intracellular trafficking of SPAK by SORLA is essential for NKCC2 activation and for renal ion handling.     

Experimental pig yersiniosis to assess attenuation of Yersinia enterocolitica O:8 mutant strains

An experimental oral pig model was used to assess the pathogenic and immunogenic potential of Yersinia enterocolitica serotype O:8 wild-type strain 8081-L2 and its lipopolysaccharide (LPS) mutant derivatives: a spontaneous rough mutant 8081-R2, strain 8081-DeltawzzGB expressing O-antigen with uncontrolled chain lengths, and strain 8081-wbcEGB expressing semirough LPS with only one O-unit. Microbiological and immunological parameters of the infected pigs were followed from day 7 to 60 postinfection. The wild-type and all LPS mutant strains persisted in the lymphoid tissue of tonsils and small intestines, causing asymptomatic infection without any pathological changes. Although the pig is known as a reservoir of Yersiniae, a precise analysis of pathogenic and immunogenic parameters based on different in vitro tests (hematological response, killing ability of leukocytes and blood sera, antibody response, hydrogen peroxide production by macrophages, classical and alternative pathways of complement activation), revealed significant attenuation in the pathogenicity of the LPS mutant strains but not the loss of immunogenic potential. In comparison with the other strains, strain 8081-DeltawzzGB demonstrated more continuous leucocytosis with monocytosis, higher invasive potential, significant activation of hydrogen peroxide production by macrophages and an effective immunoglobulin G immune response accompanied by relevant histological immunomorphological rearrangements.