Perinatal and long-term effects of maternal uterine artery adenoviral VEGF-A165 gene therapy in the growth-restricted guinea pig fetus.

Uterine artery application of adenoviral vascular endothelial growth factor A165 (Ad.VEGF-A165) gene therapy increases uterine blood flow and fetal growth in experimental animals with fetal growth restriction (FGR). Whether Ad.VEGF-A165 reduces lifelong cardiovascular disease risk imposed by FGR remains unknown. Here, pregnant guinea pigs fed 70% normal food intake to induce FGR received Ad.VEGF-A165 (1×1010 viral particles, n = 15) or vehicle ( n = 10), delivered to the external surface of the uterine arteries, in midpregnancy. Ad libitum-fed controls received vehicle only ( n = 14). Litter size, gestation length, and perinatal mortality were similar in control, untreated FGR, and FGR+Ad.VEGF-A165 animals. When compared with controls, birth weight was lower in male but higher in female pups following maternal nutrient restriction, whereas both male and female FGR+Ad.VEGF-A165 pups were heavier than untreated FGR pups ( P < 0.05, ANOVA). Postnatal weight gain was 10-20% greater in female FGR+Ad.VEGF-A165 than in untreated FGR pups, depending on age, although neither group differed from controls. Maternal nutrient restriction reduced heart weight in adult female offspring irrespective of Ad.VEGF-A165 treatment but did not alter ventricular wall thickness. In males, postnatal weight gain and heart morphology were not affected by maternal treatment. Neither systolic, diastolic, mean arterial pressure, adrenal weight, nor basal or challenged plasma cortisol were affected by maternal undernutrition or Ad.VEGF-A165 in either sex. Therefore, increased fetal growth conferred by maternal uterine artery Ad.VEGF-A165 is sustained postnatally in FGR female guinea pigs. In this study, we did not find evidence for an effect of maternal nutrient restriction or Ad.VEGF-A165 therapy on adult offspring blood pressure.

Autoantibodies against complement components in systemic lupus erythematosus - role in the pathogenesis and clinical manifestations.

Many complement structures and a number of additional factors, i.e. autoantibodies, receptors, hormones and cytokines, are implicated in the complex pathogenesis of systemic lupus erythematosus. Genetic defects in the complement as well as functional deficiency due to antibodies against its components lead to different pathological conditions, usually clinically presented. Among them hypocomplementemic urticarial vasculitis, different types of glomerulonephritis as dense deposit disease, IgA nephropathy, atypical haemolytic uremic syndrome and lupus nephritis are very common. These antibodies cause conformational changes leading to pathological activation or inhibition of complement with organ damage and/or limited capacity of the immune system to clear immune complexes and apoptotic debris. Finally, we summarize the role of complement antibodies in the pathogenesis of systemic lupus erythematosus and discuss the mechanism of some related clinical conditions such as infections, thyroiditis, thrombosis, acquired von Willebrand disease, etc.

MBL2 polymorphisms - manifestations in Bulgarian patients with adult dermatomyositis and systemic lupus erythematosus.

Deficiency in some complement factors is known to cause both systemic lupus erythematosus (SLE) and dermatomyositis (DM). Mannose-binding lectin (MBL) is a recognition molecule of the lectin pathway, and its low levels are reported to influence some autoimmune diseases. Furthermore, MBL2 polymorphisms have been described associated with low MBL serum levels due to impaired MBL structure and function. This is a pilot study to investigate the role of MBL2-550G/C (H/L), -221G/C (Y/X), Arg52Cys (D), Gly54Asp (B), Gly57Glu (C) polymorphisms and MBL serum levels as a risk factor for a development of adult DM and SLE in Bulgarian patients. None of the studied MBL2 polymorphisms appeared associated with the diseases investigated. However, we have found an increased OR of MBL2-221XY genotype in the patients with SLE (OR 1.64, 95%CI 0.77-3.52). MBL2 polymorphisms seemed to affect MBL serum levels and to be associated with the clinical features although none of the associations remained statistically significant after Bonferroni correction. The-550L allele showed an association with electromyography findings in patients with DM. The-221XY genotype was associated with photosensitivity in patients with SLE. The 54AB genotype showed an association with malar rash in patients with SLE, but it appeared decreased among SLE patients with ANA. In conclusion, our results suggest that the MBL2 polymorphisms have rather a disease modifying role and they are not associated with the disease susceptibility in adult DM and SLE among Bulgarian patients.

The self-reported density of truck traffic on residential streets and the impact on asthma, hay fever and eczema in young adolescents.

BACKGROUND: Conflicting results have been reported, mostly in developed countries, on the relationship between exposure to traffic and allergic diseases. This study aims to examine the impact of truck traffic on asthma, rhinitis and eczema in early adolescence in Skopje, the capital of the Republic of Macedonia, as a developing country with a lower middle rate of high truck traffic exposure and low prevalence rates of allergic diseases. METHODS: Self-reported data was used, obtained through the International Study of Asthma and Allergies in Childhood Phase 3 written questionnaires, from 3026 adolescents aged 13-14 years from Skopje. Truck traffic density on the street of residence on weekdays was correlated to current and ever-diagnosed asthma, rhinitis and eczema by odds ratios (OR, 95% CI) in binary logistic regression, with and without adjustments for potential confounding factors separately and for their joint effect. RESULTS: A positive association of truck traffic density appeared to be limited to current dry night cough (aOR: 1.63; 1.07-2.47; aOR: 2.17; 1.40-3.35; and aOR: 2.33; 1.43-3.79 for truck traffic seldom, frequently through the day, and almost the whole day, respectively) with an exposure-response relationship and to current wheeze only for truck traffic almost the whole day (aOR: 1.87; 1.02-3.42). CONCLUSION: The findings suggest an aggravating effect of truck traffic on current asthma symptoms, but not on asthma, allergic rhinitis and eczema diagnoses. It seems that it probably has an impact as a direct respiratory irritant in early adolescence.

Temperature response of sucrose palmitate solutions: Role of ratio between monoesters and diesters.

HYPOTHESIS: Aqueous solutions of long-chain water-soluble sucrose ester surfactants exhibit non-trivial response to temperature variations, revealing a peak in viscosity around 40-50 °C. While previous investigations have explored the structures within sucrose stearate systems at various constant temperatures, a comprehensive understanding of the entire temperature dependence and the underlying molecular factors, contributing to this phenomenon is currently missing. EXPERIMENTS: Temperature dependent properties and supramolecular structures formed in aqueous solutions of commercial sucrose palmitate were examined using SAXS/WAXS, DSC, optical microscopy, rheological measurements, NMR, and cryo-TEM. FINDINGS: The underlying mechanism governing this unusual behavior is revealed and is shown to relate to the mono- to di-esters ratio in the solutions. Solutions primarily containing sucrose monoesters (monoesters molecules ≳ 98% of all surfactant molecules) exhibit behavior typical of nonionic surfactants, with minimal changes with temperature. In contrast, the coexistence of mono- and di-esters results in the formation of discrete monodisperse diester particles and a network of partially fused diester particles at low temperature. As the temperature approaches the diesters' melting point, wormlike mixed micelles form, causing a viscosity peak. The height of this peak increases significantly with the diester concentration. Further temperature increase leads to fluidization of surfactant tails and formation of branched micelles, while excess diester molecules phase separate into distinct droplets.

Maternal Uterine Artery Adenoviral Vascular Endothelial Growth Factor (Ad.VEGF-A165) Gene Therapy Normalises Fetal Brain Growth and Microglial Activation in Nutrient Restricted Pregnant Guinea Pigs.

Fetal growth restriction (FGR) is associated with uteroplacental insufficiency, and neurodevelopmental and structural brain deficits in the infant. It is currently untreatable. We hypothesised that treating the maternal uterine artery with vascular endothelial growth factor adenoviral gene therapy (Ad.VEGF-A165) normalises offspring brain weight and prevents brain injury in a guinea pig model of FGR. Pregnant guinea pigs were fed a restricted diet before and after conception and received Ad.VEGF-A165 (1 × 1010 viral particles, n = 18) or vehicle (n = 18), delivered to the external surface of the uterine arteries, in mid-pregnancy. Pregnant, ad libitum-fed controls received vehicle only (n = 10). Offspring brain weight and histological indices of brain injury were assessed at term and 5-months postnatally. At term, maternal nutrient restriction reduced fetal brain weight and increased microglial ramification in all brain regions but did not alter indices of cell death, astrogliosis or myelination. Ad.VEGF-A165 increased brain weight and reduced microglial ramification in fetuses of nutrient restricted dams. In adult offspring, maternal nutrient restriction did not alter brain weight or markers of brain injury, whilst Ad.VEGF-A165 increased microglial ramification and astrogliosis in the hippocampus and thalamus, respectively. Ad.VEGF-A165 did not affect cell death or myelination in the fetal or offspring brain. Ad.VEGF-A165 normalises brain growth and markers of brain injury in guinea pig fetuses exposed to maternal nutrient restriction and may be a potential intervention to improve childhood neurodevelopmental outcomes in pregnancies complicated by FGR.

The effects of melatonin on burn-induced inflammatory responses and coagulation disorders in rats.

The aim of this study was to investigate the effects of melatonin on burn-induced inflammatory responses and coagulation disorders in a rat model. Under ether anesthesia, shaved dorsa of rats were exposed to 90 degrees C for 10 s to induce burn injury. Melatonin (10 mg/kg) was administered immediately and after 12 h. Standard coagulation tests (prothrombin activity [PA] and activated partial thromboplastin time [aPTT]), platelet number and morphology; proinflammatory markers (C-reactive protein [CRP] and fibrinogen) and lipid peroxidation marker (malondialdehyde [MDA]) levels were assayed. Thermal injury increased the levels of MDA (by 76%, P < 0.0001), CRP (by 33%, P < 0.0001), fibrinogen (4.5-fold, P < 0.0001) and PA (by 37%; P < 0.01). Changes in aPTT and platelet numbers were nonsignificant. Melatonin diminished the elevated CRP and fibrinogen levels, normalized MDA levels, platelet morphology and decreased PA. A positive association of MDA with fibrinogen and MDA with PA were noted after melatonin treatment. To conclude, melatonin as an antioxidant and anti-inflammatory agent exerted a suppressive effect on burn-induced disorders in blood coagulation and might be useful in the prevention of disseminated intravascular microthrombosis.

Protective effect of melatonin against oxidative hepatic injury after experimental thermal trauma.

Burns cause thermal injury to local tissue, trigger systemic inflammatory processes and activate lipid peroxidation, leading to multiple distant organ injury. Melatonin is a lipid- and water-soluble antioxidant and membrane stabilizer with antiinflammatory, hepatoprotective and gastroptotective properties, among others. We studied the influence of melatonin on hepatic damage induced by thermal skin injury and its possible relation to hepatic lipid peroxidative status and systemic inflammatory response. Under ether anesthesia the shaved dorsum of rats was exposed to a 90 degrees C bath for 10 s. Melatonin was administered intraperitoneally immediately after burns. Malondialdehyde (MDA), aspartate transaminase (AST) and alanine transaminase (ALT) were determined in liver and blood plasma and used as markers of oxidative status. Plasma C-reactive protein (CRP) was used as a marker of systemic inflammatory response. Thermal skin injury caused significant elevation of hepatic MDA by 48%, plasma CRP levels by 30% and plasma AST and ALT activities by 2- and 3.5-fold, respectively, in comparison with normal control rats. Treatment with melatonin (10 mg/kg) significantly inhibited the elevation in hepatic MDA and plasma CRP levels, reaching control values at 24 h. Melatonin treatment restricts the elevation of plasma AST and ALT activities (P < 0.001), which remain significantly increased as compared with controls. In conclusion, the protective effect of melatonin is likely to be due to attenuated lipid peroxidation and interference with reduced oxidative stress and inflammatory response, as evidenced by decreased hepatic MDA and plasma CRP levels.

Metabolic syndrome--neurotrophic hypothesis.

An increasing number of researchers of the metabolic syndrome assume that many mechanisms are involved in its complex pathophysiology such as an increased sympathetic activity, disorders of the hypothalamo-pituitary-adrenal axis, the action of chronic subclinical infections, proinflammatory cytokines, and the effect of adipocytokines or psychoemotional stress. An increasing body of scientific research in this field confirms the role of the neurotrophins and mastocytes in the pathogenesis of inflammatory and immune diseases. Recently it has been proved that neurotrophins and mastocytes have metabotrophic effects and take part in the carbohydrate and lipid metabolism. In the early stage of the metabolic syndrome we established a statistically significant increase in the plasma levels of the nerve growth factor. In the generalized stage the plasma levels of the neutrophines were statistically decreased in comparison to those in the healthy controls. We consider that the neurotrophin deficit is likely to play a significant pathogenic role in the development of the metabolic anthropometric and vascular manifestations of the generalized stage of MetSyn. We suggest a hypothesis for the etiopathogenesis of the metabolic syndrome based on the neuro-immuno-endocrine interactions. The specific pathogenic pathways of MetSyn development include: (1) increased tissue and plasma levels of proinflammatory cytokines Interleukin-1(IL-1), Interleukin-6 (IL-6 ) and tumor necrosis factor - alpha (TNF-alpha) caused by inflammatory and/or emotional distress; (2) increased plasma levels of neurotrophin - nerve growth factor (NGF) caused by the high IL-1, IL-6 and TNFalpha levels; (3) high plasma levels of NGF which enhance activation of: the autonomous nerve system--vegetodystonia (disbalance of neurotransmitters); Neuropeptide Y (NPY)--enhanced feeding, obesity and increased leptin plasma levels; hypothalamo-pituitary-adrenal axis--increased corticotropin-releasing hormone (CRH) and cortisol (hormonal disbalance); immune cells--increased number and degranulation of mastocytes (MC)--immunological disbalance; (4) as a result of 1-3 insulin resistance is exhibited leading to diabetes mellitus. The hypothesis is confirmed by results obtained after 6-month nonsteroid anti-inflammatory treatment of patients with MetSyn. These results are reported in a separate publication.

DUOX1 silencing in lung cancer promotes EMT, cancer stem cell characteristics and invasive properties.

Dual oxidase 1 (DUOX1) is an oxidant-generating enzyme within the airway epithelium that participates in innate airway host defense and epithelial homeostasis. Recent studies indicate that DUOX1 is suppressed in lung cancers by epigenetic silencing, although the importance of DUOX1 silencing in lung cancer development or progression is unknown. Here we show that loss of DUOX1 expression in a panel of lung cancer cell lines is strongly associated with loss of the epithelial marker E-cadherin. Moreover, RNAi-mediated DUOX1 silencing in lung epithelial cells and the cancer cell line NCI-H292 was found to result in loss of epithelial characteristics/molecular features (altered morphology, reduced barrier function and loss of E-cadherin) and increased mesenchymal features (increased migration, anchorage-independent growth and gain of vimentin/collagen), suggesting a direct contribution of DUOX1 silencing to epithelial-to-mesenchymal transition (EMT), an important feature of metastatic cancer. Conversely, overexpression of DUOX1 in A549 cells was capable of reversing EMT features. DUOX1 silencing in H292 cells also led to enhanced resistance to epidermal growth factor receptor tyrosine kinase inhibitors such as erlotinib, and enhanced levels of cancer stem cell (CSC) markers CD133 and ALDH1. Furthermore, acquired resistance of H292 cells to erlotinib resulted in enhanced EMT and CSC features, as well as loss of DUOX1. Finally, compared with control H292 cells, H292-shDUOX1 cells displayed enhanced invasive features in vitro and in vivo. Collectively, our findings indicate that DUOX1 silencing in lung epithelial cancer cells promotes features of EMT, and may be strongly associated with invasive and metastatic lung cancer.

Evidence for an Elongation/Reduction/C1-Elimination Pathway in the Biosynthesis of n-Heptane in Xylem of Jeffrey Pine.

The biosynthetic pathway to n-heptane was investigated by examining the effect of the [beta]-keto acyl-acyl carrier protein synthase inhibitor (2R,3S)-2,3-epoxy-4-oxo-7E,10E-dodecadienamide (cerulenin), a thiol reagent ([beta]-mercaptoethanol), and an aldehydetrapping reagent (hydroxylamine) on the biosynthesis of n-[14C]heptane and putative intermediates in xylem sections of Jeffrey pine (Pinus jeffreyi Grev.& Balf.) incubated with [14C]acetate. Cerulenin inhibited C18 fatty acid biosynthesis but had relatively little effect on radiolabel incorporation into C8 fatty acyl groups and n-heptane. [beta]-Mercaptoethanol inhibited n-heptane biosynthesis, with a corresponding accumulation of radiolabel into both octanal and 1-octanol, whereas hydroxylamine inhibited both n-heptane and 1-octanol biosynthesis, with radiolabel accumulation in octyl oximes. [14C]Octanal was converted to both n-heptane and 1-octanol when incubated with xylem sections, whereas [14C]1-octanol was converted to octanal and n-heptane in a hydroxylamine-sensitive reaction. These results suggest a pathway for the biosynthesis of n-heptane whereby acetate is polymerized via a typical fatty acid synthase reaction sequence to yield a C8 thioester, which subsequently undergoes a two-electron reduction to generate a free thiol and octanal, the latter of which alternately undergoes an additional, reversible reduction to form 1-octanol or loss of C1 to generate n-heptane.

The role of different strain backgrounds in bacterial endotoxin-mediated sensitization to neonatal hypoxic-ischemic brain damage.

Genetic background is known to influence the outcome in mouse models of human disease, and previous experimental studies have shown strain variability in the neonatal mouse model of hypoxia-ischemia. To further map out this variability, we compared five commonly used mouse strains: C57BL/6, 129SVJ, BALB/c, CD1 and FVB in a pure hypoxic-ischemic setup and following pre-sensitization with lipopolysaccharide (LPS). Postnatal day 7 pups were subjected to unilateral carotid artery occlusion followed by continuous 30 min 8% oxygen exposure at 36 °C. Twelve hours prior, a third of the pups received a single intraperitoneal LPS (0.6 µg/g) or a saline (vehicle) administration, respectively; a further third underwent hypoxia-ischemia alone without preceding injection. Both C57BL/6 and 129SVJ strains showed minimal response to 30min hypoxia-ischemia alone, BALB/c demonstrated a moderate response, and both CD1 and FVB revealed the highest brain damage. LPS pre-sensitization led to substantial increase in overall brain infarction, microglial and astrocyte response and cell death in four of the five strains, with exception of BALB/c that only showed a significant effect with terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). Saline administration prior to hypoxia-ischemia resulted in an increase in inflammatory-associated markers, particularly in the astroglial activation of C57BL/6 mice, and in combined microglial activation and neuronal cell loss in FVB mice. Finally, two of the four strongly affected strains--C57BL/6 and CD1--revealed pronounced contralateral astrogliosis with a neuroanatomical localization similar to that observed on the occluded hemisphere. Overall, the current findings demonstrate strain differences in response to hypoxia-ischemia alone, to stress associated with vehicle injection, and to LPS-mediated pre-sensitization, which partially explains the high variability seen in the neonatal mouse models of hypoxia-ischemia. These results can be useful in future studies of fetal/neonatal response to inflammation and reduced oxygen-blood supply.

Microglial major histocompatibility complex glycoprotein-1 in the axotomized facial motor nucleus: regulation and role of tumor necrosis factor receptors 1 and 2.

Presentation of antigen is key to the development of the immune response, mediated by association of antigen with major histocompatibility complex glycoproteins abbreviated as MHC1 and MHC2. In the current study, we examined the regulation of MHC1 in the brain after facial axotomy. The normal facial motor nucleus showed no immunoreactivity for MHC1 (MHC1-IR). Transection of the facial nerve led to a strong and selective up-regulation of MHC1-IR on the microglia in the affected nucleus, beginning at day 2 and reaching a maximum 14 days after axotomy, coinciding with a peak influx of the T lymphocytes that express CD8, the lymphocyte coreceptor for MHC1. Specificity of the MHC1 staining was confirmed in beta2-microglobulin-deficient mice, which lack normal cell surface MHC1-IR. MHC1-IR was particularly strong on phagocytic microglia, induced by delayed neuronal cell death, and correlated with the induction of mRNA for tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and interferon-gamma and the influx of T lymphocytes. Mice with severe combined immunodeficiency (scid), lacking T and B cells, showed an increase in the number of MHC1-positive nodules but no significant effect on overall MHC1-IR. Transgenic deletion of the IL1 receptor type I, or the interferon-gamma receptor type 1 subunit, did not affect the microglial MHC1-IR. However, a combined deletion of TNF receptors 1 and 2 (TNFR1&2-KO) led to a decrease in microglial MHC1-IR and to a striking absence of the phagocytic microglial nodules. Deletion of TNFR2 (p75) did not have an effect; deletion of TNFR1 (p55) reduced the diffuse microglial staining for MHC1-IR but did not abolish the MHC1(+) microglial nodules. In summary, neural injury leads to the induction of MHC1-IR on the activated, phagocytic microglia. This induction of MHC1 precedes the interaction with the immune system, at least in the facial motor nucleus model. Finally, the impaired induction of these molecules, up to now, only in the TNFR-deficient mice underscores the central role of TNF in the immune activation of the injured nervous system.

Adipobiology of disease: adipokines and adipokine-targeted pharmacology.

In recent years, the simple paradigm of adipose tissue as merely a fat store is rapidly evolving into a complex paradigm of this tissue as multipotential secretory organ, partitioned into a few large depots, including visceral and subcutaneous location, and many small depots, associated with a variety of organs in the human body. The major secretory compartment of adipose tissue consists of adipocytes, fibroblasts, and mast cells. These cells, using endocrine, paracrine and autocrine pathways, secrete multiple bioactive molecules, conceptualized as adipokines or adipocytokines. This review examines current information in adipobiology of various diseases besides obesity and related diseases such as type 2 diabetes, metabolic syndrome, and cardiovascular disease. Finally, we emphasize the possibilities for adipokine-targeted pharmacology in adiponectin (Acrp30, apM1, AdipoQ, GBP28), angiotensin II, estrogens, nerve growth factor, tumor necrosis factor-alpha, and also adipose mast cells.

Treatment with oxoglaucine can enhance host resistance to Candida albicans infection of mice with adjuvant arthritis.

The alkaloid oxoglaucine reduced CD4+ cell clones in adult mice and decreased CD4+, CD8+ and Ig+ levels in newborn mice. It prevented the increase of CD8+ and Ig+ clones induced by Candida albicans (C. albicans) in adult mice. TNF-alpha serum accumulation was inhibited by oxoglaucine in C. albicans infection and adjuvant arthritis. Treatment with oxoglaucine of arthritic mice, followed by inoculation with C. albicans enhanced the host resistance against the pathogen.

IMMUNOSUPPRESSION AND RECOVERY OF DRUG-IMPAIRED HOST RESISTANCE AGAINST CANDIDA ALBICANS INFECTION BY OXOGLAUCINE.

The immunosuppressive action of aporphinoid alkaloid oxoglaucine was studied in experimental Candida albicans (C. albicans) infection in mice. The alkaloid augmented host resistance to pathogen applied to mice (6-8 weeks of age) at a low dose of 2 mg kg(-1)in 3 days and impaired it at a high dose of 10 mg kg(-1). The suppressive activity observed under the latter schedule correlated with the inhibited proliferative response of splenic cells to mitogens and with decreased popliteal lymph node (PLN) reaction to C. albicans. Treatment of mice with oxoglaucine (at the age of 5 days) at a dose of 5 mg kg(-1)in 3 consecutive days increased the susceptibility to Candida inoculation at the age of 6 weeks. Delayed type hypersensitivity (DTH) response to C. albicans was enhanced after pretreatment of adult mice and was suppressed after administration to newborn mice. Long-time treatment (10 days) with oxoglaucine, cyclophoshamide or prednisolone at a dose of 10 mg kg(-1)increased the rate of mortality of Candida -infected mice. Combined pretreatment of mice with cyclophosphamide or prednisolone (5 days at a dose of 5 mg kg(-1)) followed by oxoglaucine (5 days at a dose of 5 mg kg(-1)), prolonged the survival of infected mice. 2000 Academic Press@p$hr Copyright 2000 Academic Press.

NGF, BDNF, leptin, and mast cells in human coronary atherosclerosis and metabolic syndrome.

While multiple growth factor, cytokines, and immune cells are identified in atherosclerotic lesions, as well as an essential nonneuronal function of neurotrophins implicated in cardiovascular tissue development and in lipid and glucose metabolism, the role of the neurotrophins NGF and BDNF and also the adipokine leptin in human coronary atherosclerosis and related disorders, such as metabolic syndrome, remains unclear. Here we report that (i) both the amount and the immunoreactivity of NGF was reduced and the expression of p75NGF receptor and the number of mast cell increased in human atherosclerotic coronary arteries (n = 12) compared with control specimens (n = 9) obtained from autopsy cases, and (ii) NGF and BDNF plasma levels were reduced in patients with metabolic syndrome (n = 23) compared with control subjects (n = 10). Also, in metabolic syndrome patients, a positive correlation between the plasma leptin levels and the number of adipose tissue mast cells was found, suggesting that leptin may be a novel adipoimmune mediator. Altogether, the results provide the first correlative evidence for the potential involvement of NGF, BDNF, leptin, and mast cells in human coronary atherosclerosis and metabolic syndrome, implying neuroimmune and adipoimmune pathways in the pathobiology of these cardiovascular disorders.

Changes in the aerobic capacity and ultrastructure of skeletal muscles and myocardium of endurance training rats induced by specialized food supplements with different compositions.

In order to find how the aerobic capacity and the ultrastructure of the skeletal and heart muscle cells of endurance training rats change as a result of adding two dietary supplements with different compositions to their diet (our own dietary formula specially designed for endurance sports and Aufbau Konzentrat, a product of Pfrimmer, Germany)--we carried out an experiment with 16 Wistar rats divided in two groups. The animals from group A received a food mixture containing 76% standard rat chow and 24% of the endurance training supplement, and those from group B--a food mixture of 76% standard rat chow and 24% Aufbau Konzentrat. Both supplements contained a high-quality protein, but those designed for endurance training was of higher fat and lower carbohydrate content. The animals were subjected to training on a treadmill at submaximal loading for 8 weeks. Initially and at the end of the experiment (week 9) VO2max-test was performed and VO2max and RQ variables of the treated animals were evaluated. At the end of the experiment the maximum oxygen intake was greater in group A animals than that in group B rats--85.06 +/- 8.81 and 75.77 +/- 4.91 ml O2.kg-1.min-1, respectively. After eight weeks a material from m. gastrocnemius and from the left cardiac ventricle of all animals was obtained and treated for transimmersion electron microscopy. Morphological data for more intensive oxidative processes in the gastrocnemius muscle and myocardium of group A animals were evident. The greater number of mitochondria and the higher cristae density observed in the gastrocnemius muscle cells indicated a better aerobic adaptation induced by the training and the intake of the specially designed supplement for endurance sports. Within an 8-week period of submaximal training using the specially adapted food formula resulted in a better structural basis than Aufbau Konzentrat produced in the skeletal and heart muscle cells for improvement of the aerobic capacity.

Complications of automated spring fired biopsy gun technique. A retrospective analysis of 230 cases.

BACKGROUND: Renal biopsy (RB) is a procedure which allows obtaining kidney tissue for a morphological diagnosis. As an invasive procedure, RB could lead to some complications. The aim of the study was to assess the incidence and the severity of various complications of renal biopsy(RB) performed by automated spring fired biopsy gun with needles 16 and 18G. MATERIAL AND METHODS: A total number of 230 RB obtained from patients over 18 years of age, within the period from January 2009 to July 2013, were analyzed retrospectively. All RB were performed by a standartized technique using an automated spring fired biopsy gun, with 16, and 18 G disposable needle. The average age of the patients was 45.5 ± 22.3 years, 119 male and 111 female. As many as 170 biopsies were performed with 16 G needle and 60 with 18G. The assessment of the early post-biopsy complications was done by renal ultrasound (US). RESULTS: Of all RB, 92.7% were successfully performed. In 60% of the failed RB 18 G needle was used. No post-biopsy complications were noted in 93.4%. Hematomas were observed in 15 out of 230 cases (6.6%). Of those, 80% were assessed as minor hematomas (12/15) with an average size of 20.4mm ± 11mm and they appeared to be asymptomatic.While 8 cases (66%) of all 12 minor hematomas were obtained by two passes (66%), only 4 cases (34%) of hematoma were observed after one pass <0.01. The cases of hematomas obtained with 16 G needle were significantly higher than those in the group with 18G (8 to 4, p<0,05). However, when the number of hematomas in both groups referred to the number of all biopsies in the corresponding group, no significant difference of this complication was observed, p > 0.05. The incidence of major complications was 1.3%. CONCLUSION: The optimal period for US examination of the kidneys for early diagnosis of complications is up to 24 hours after RB. The experience of the physician performing the procedure is of great importance for reducing the risk of complications as well as the RB technique used. An automated spring fired biopsy gun with needle 16G is recommended.

NGS nominated CELA1, HSPG2, and KCNK5 as candidate genes for predisposition to Balkan endemic nephropathy.

Balkan endemic nephropathy (BEN) is a familial chronic tubulointerstitial disease with insidious onset and slow progression leading to terminal renal failure. The results of molecular biological investigations propose that BEN is a multifactorial disease with genetic predisposition to environmental risk agents. Exome sequencing of 22 000 genes with Illumina Nextera Exome Enrichment Kit was performed on 22 DNA samples (11 Bulgarian patients and 11 Serbian patients). Software analysis was performed via NextGene, Provean, and PolyPhen. The frequency of all annotated genetic variants with deleterious/damaging effect was compared with those of European populations. Then we focused on nonannotated variants (with no data available about them and not found in healthy Bulgarian controls). There is no statistically significant difference between annotated variants in BEN patients and European populations. From nonannotated variants with more than 40% frequency in both patients' groups, we nominated 3 genes with possible deleterious/damaging variants--CELA1, HSPG2, and KCNK5. Mutant genes (CELA1, HSPG2, and KCNK5) in BEN patients encode proteins involved in basement membrane/extracellular matrix and vascular tone, tightly connected to process of angiogenesis. We suggest that an abnormal process of angiogenesis plays a key role in the molecular pathogenesis of BEN.