Effects of quinazolinones on Balb/C mice embryonic livers

Heterocyclic compounds such as quinazolinones have variety of biological and pharmacological properties (anticancer, antiinflammatory, antimicrobial, antimalaria, etc.). Effects of two new quinazolinones viz., 4(3H)-quinazolinone-2-propyl-2-phenylethyl (QPPE) and 4(3H)quinazolinone-2-ethyl-2-phenylethyl (QEPE) were investigated on Balb/C mice embryos livers-the major organ of metabolism and detoxification of drugs and toxins. Histological and pathological studies demonstrated QPPE and QEPE as producers of toxic metabolites after biotransformation, creating necrosis, fatty changes, increase in the number of band cells, hepatocytes' diameters and alkaline phosphatase, in addition to sinusoid dilation, hemorrhages and hyperemia. Transmission electron micrographs showed lipid droplets in hepatocytes' cytoplasm, necrosis, vacuolization, cytoplasm disintegration, disfigured and swollen mitochondria, irregular and abnormal nuclei, nuclei with heterochromatin, condensed chromatins, myelin figures and autophages in injured hepatocytes. In conclusion, QPPE and QEPE make toxic components after biotransformation injuring membranes and creating inflammatory reactions. They also disturb metabolism of lipids pathways and cause the appearances of lipid droplets in hepatocytes.     

Activity and turnover of eosinophil and neutrophil granulocytes are altered in visceral leishmaniasis

Visceral leishmaniasis (VL) is a health issue in Sudan. Our aim was to investigate the involvement of eosinophils and neutrophils in VL by serum and plasma measurements of eosinophil cationic protein (ECP) and myeloperoxidase (MPO) and some key cytokines and chemokines. Blood was collected from 125 VL patients and 181 healthy Sudanese controls from the same rural area. Results showed reduced eosinophil and neutrophil counts in the VL group (P=0.0001 and P=0.002, respectively). Serum-ECP levels were higher in the controls (P<0.0001), while plasma MPO levels were higher in the VL group (P<0.0001). Levels of IL-5, granulocyte macrophage-colony stimulating factor (GM-CSF) and IL-17 were increased among the VL group (P<0.0001, P=0.017 and P=0.03, respectively), whereas eotaxin and IL-8 levels were reduced (P<0.0001 and P=0.002, respectively). Positive correlations were found between IL-8 and ECP/MPO (P<0.0001). We conclude that eosinophil and neutrophil turnover and activity are increased in subjects in rural areas of Sudan. In VL the turnover was further increased, but the relatively low secretory activity of eosinophils and neutrophils in VL may relate to the reduced production and availability of the chemokines eotaxin and IL-8. The combined assay of ECP and MPO in serum and plasma provides further insight into the mechanisms of eosinophil and neutrophil involvement in disease and constitutes a novel approach to the study of disease processes.     

Alginate microspheres encapsulated with autoclaved Leishmania major (ALM) and CpG-ODN induced partial protection and enhanced immune response against murine model of leishmaniasis

A suitable adjuvant and delivery system are needed to enhance efficacy of vaccines against leishmaniasis. In this study, alginate microspheres as an antigen delivery system and CpG-ODN as an immunoadjuvant were used to enhance immune response and induce protection against an experimental autoclaved Leishmania major (ALM) vaccine. Alginate microspheres were prepared by an emulsification technique and the characteristics of the preparation such as size, encapsulation efficiency and release profile of encapsulates were studied. Mean diameter of microspheres was determined using SEM (Scanning Electron Microscopy) and particle size analyzer. The encapsulation efficiency was determined using Lowry protein assay method. The integrity of ALM antigens was assessed using SDS–PAGE. Mean diameter of microspheres was 1.8 ± 1.0 μm. BALB/c mice were immunized three times in 3-weeks intervals with ALM + CpG-ODN loaded microspheres [(ALM + CpG)_ALG], ALM encapsulated alginate microspheres [(ALM)_ALG], (ALM)_ALG + CpG, ALM + CpG, ALM alone or PBS. The intensity of infection induced by L. major challenge was assessed by measuring size of footpad swelling. The strongest protection was observed in group of mice immunized with (ALM + CpG)_ALG. The groups of mice received (ALM + CpG)_ALG, (ALM)_ALG + CpG, (ALM)_ALG and ALM + CpG were also showed a significantly (P < 0.05) smaller footpad swelling compared with the group that received either ALM alone or PBS. The mice immunized with (ALM + CpG)_ALG or ALM + CpG showed the significantly (P < 0.05) highest IgG2a/IgG1 ratio. The IFN-γ level was significantly (P < 0.0001) highest in group of mice immunized with either (ALM)_ALG + CpG or ALM + CpG. It is concluded that alginate microspheres and CpG-ODN adjuvant when are used simultaneously induced protection and enhanced immune response against ALM antigen.     

Lipase maturation factor 1 is required for endothelial lipase activity

Lipase maturation factor 1 (Lmf1) is an endoplasmic reticulum (ER) membrane protein involved in the posttranslational folding and/or assembly of lipoprotein lipase (LPL) and hepatic lipase (HL) into active enzymes. Mutations in Lmf1 are associated with diminished LPL and HL activities ("combined lipase deficiency") and result in severe hypertriglyceridemia in mice as well as in human subjects. Here, we investigate whether endothelial lipase (EL) also requires Lmf1 to attain enzymatic activity. We demonstrate that cells harboring a (cld) loss-of-function mutation in the Lmf1 gene are unable to generate active EL, but they regain this capacity after reconstitution with the Lmf1 wild type. Furthermore, we show that cellular EL copurifies with Lmf1, indicating their physical interaction in the ER. Finally, we determined that post-heparin phospholipase activity in a patient with the LMF1(W464X) mutation is reduced by more than 95% compared with that in controls. Thus, our study indicates that EL is critically dependent on Lmf1 for its maturation in the ER and demonstrates that Lmf1 is a required factor for all three vascular lipases, LPL, HL, and EL.     

Ugan古河道胡杨可培养内生细菌的多样性

摘要：【目的】为了了解塔河废弃古河道胡杨可培养内生细菌的多样性。【方法】从2棵胡杨树干部抽出其内存液,采用三种不同的培养基对样品的内生细菌进行了分离纯化;对它们进行16S rDNA测定和系统进化分析。【结果】分离纯化不同表型的细菌62株,对它们的16S rDNA序列分析表明,62株菌分别属于四个大类群;厚壁菌门（Firmicutes）、放线菌门(Actinobacteria)、α-变形菌纲(Alpha Proteobacteria) 、γ-变形菌纲(Gamma Proteobacteria),18个属,32个种;芽孢杆菌属和假单胞菌属是胡杨可培养内生细菌的优势细菌种群,它们分别占已测种群的40.32%、16.13%。其中菌株KTH-63为葡萄球菌科的潜在的新属新种,它与最近源菌株的16S rDNA序列相似率为92.491%;9株菌KLH-21、KLH-1、KTH-8、KTH-14、KNA-26、KLH-18、KTH-20、KNA-3、KLH-25是潜在的新种（16S rDNA相似率为96.089 %-97.769 %）,胡杨树干内存液中潜在新种的发现率高达总分离检测菌株的16.13 % 。本研究获得的胡杨可培养内生细菌的群落结构数据给植物内生细菌新增了10个属,18个种。【结论】胡杨具有多样性极其丰富的可培养内生细菌菌种资源,土著新种的发现频率超出了预期,胡杨可培养内生细菌的群落结构极大地刷新了植物内生细菌的种群记录,极具进一步发掘的潜力。     

Rhizobial strain involvement in symbiosis efficiency of chickpea–rhizobia under drought stress: plant growth,nitrogen fixation and antioxidant enzyme activities

Chickpea plants were inoculated with two strains of Mesorhizobium ciceri: local strain (C-15) and non-local strain (CP-36) in order to evaluate plant growth parameters, activities of nitrogenase and antioxidant enzymes under drought stress as well as control condition within 15 days of imposition of drought stress. Biomass production, nodulation, nitrogen fixation and antioxidant enzyme activities under drought condition were compared. Under control condition, symbiotic efficiency in symbiosis formed by C-15 was higher than that in symbiosis derived by CP-36. Although drought stress decreased shoot dry weight, root dry weight, nodule dry weight and nitrogen fixation in both symbioses, the rate of decline in plants inoculated with CP-36 was higher than that in symbiosis chickpea with C-15. Therefore, symbioses showed different tolerance level under drought condition which was essentially correlated with symbiotic performance at non-stressful conditions. Under drought stress, nodular peroxidase (POX) activity increased in both symbioses but was higher in nodules produced by C-15. Ascorbate peroxidase (APX) increased significantly in nodules of symbiosis of chickpea with C-15. Catalase (CAT) and glutation reductase (GR) declined in both symbioses which decline extent in symbiosis with C-15 was lower than that in the nodules of CP-36. These results suggested contribution of rhizobial partner in enhancing the tolerance of symbioses to drought stress, which was related with the increase of antioxidant enzyme activities (APX and POX) under drought conditions.     

An l-Arginine supplement improves broiler hypertensive response and gut function in broiler chickens reared at high altitude

An experiment was carried out to examine the effects of supplemental dietary arginine (ARG) on growth, hypertensive response, and gut function in broilers reared at high altitude (2,100 m). A total of 120 day-old male broilers (Cobb 500) were divided equally into two treatment groups. Treatments included a control basal diet composed of corn and soybean meal and an experimental diet to which an l-ARG supplement was added at 10 g/kg. The trial lasted for 42 days. There were no treatment differences with regard to feed intake, body weight gain, or feed conversion ratio. However ARG supplementation did increase the plasma concentration of nitric oxide, a potent vasodilator (P?<?0.05), and attenuated indices of pulmonary hypertension as reflected by reductions in the hematocrit and the right to total ventricular weight ratio (P?<?0.05). Significantly enhanced intestinal mucosal development was observed in broilers receiving ARG supplement when compared with controls (P?<?0.05), suggesting that ARG supplementation increased the absorptive surface area of the jejunum and ileum. In conclusion, broiler diets supplemented with ARG beneficially improved pulmonary hemodynamics and appeared to enhance gut function.     

Dendritic Cell IL-1α and IL-1β Are Polyubiquitinated and Degraded by the Proteasome

IL-1α and β are key players in the innate immune system. The secretion of these cytokines by dendritic cells (DC) is integral to the development of proinflammatory responses. These cytokines are not secreted via the classical secretory pathway. Instead, 2 independent processes are required; an initial signal to induce up-regulation of the precursor pro-IL-1α and -β, and a second signal to drive cleavage and consequent secretion. Pro-IL-1α and -β are both cytosolic and thus, are potentially subject to post-translational modifications. These modifications may, in turn, have a functional outcome in the context of IL-1α and -β secretion and hence inflammation. We report here that IL-1α and -β were degraded intracellularly in murine bone marrow-derived DC and that this degradation was dependent on active cellular processes. In addition, we demonstrate that degradation was ablated when the proteasome was inhibited, whereas autophagy did not appear to play a major role. Furthermore, inhibition of the proteasome led to an accumulation of polyubiquitinated IL-1α and -β, indicating that IL-1α and -β were polyubiquitinated prior to proteasomal degradation. Finally, our investigations suggest that polyubiquitination and proteasomal degradation are not continuous processes but instead are up-regulated following DC activation. Overall, these data highlight that IL-1α and -β polyubiquitination and proteasomal degradation are central mechanisms in the regulation of intracellular IL-1 levels in DC.     

The Spatiotemporal Dynamics of Chromatin Protein HP1α Is Essential for Accurate Chromosome Segregation during Cell Division

Heterochromatin protein 1α (HP1α) is involved in regulation of chromatin plasticity, DNA damage repair, and centromere dynamics. HP1α detects histone dimethylation and trimethylation of Lys-9 via its chromodomain. HP1α localizes to heterochromatin in interphase cells but is liberated from chromosomal arms at the onset of mitosis. However, the structural determinants required for HP1α localization in interphase and the regulation of HP1α dynamics have remained elusive. Here we show that centromeric localization of HP1α depends on histone H3 Lys-9 trimethyltransferase SUV39H1 activity in interphase but not in mitotic cells. Surprisingly, HP1α liberates from chromosome arms in early mitosis. To test the role of this dissociation, we engineered an HP1α construct that persistently localizes to chromosome arms. Interestingly, persistent localization of HP1α to chromosome arms perturbs accurate kinetochore-microtubule attachment due to an aberrant distribution of chromosome passenger complex and Sgo1 from centromeres to chromosome arms that prevents resolution of sister chromatids. Further analyses showed that Mis14 and perhaps other PXVXL-containing proteins are involved in directing localization of HP1α to the centromere in mitosis. Taken together, our data suggest a model in which spatiotemporal dynamics of HP1α localization to centromere is governed by two distinct structural determinants. These findings reveal a previously unrecognized but essential link between HP1α-interacting molecular dynamics and chromosome plasticity in promoting accurate cell division.