Hypertriglyceridemia is exacerbated by slow lipolysis of triacylglycerol-rich lipoproteins in fed but not fasted streptozotocin diabetic rats.

Hydrolysis by endothelial lipases of triacylglycerol-rich lipoproteins of diabetic origin were compared to lipoproteins of non-diabetic origin. The plasma lipoprotein fraction of density < 1.006 g/ml, including chylomicrons and VLDL, were incubated in vitro with post-heparin plasma (PHP) lipases. The lipoproteins of diabetic origin were hydrolysed at a significantly slower rate than lipoproteins from normal rats by the lipoprotein lipase component of PHP. However, if rats were fasted for 16 h prior to lipoprotein recovery, no differences in rates of VLDL hydrolysis were observed. Slower hydrolysis of lipoproteins of diabetic origin reflected a decrease in the apolipoprotein CII/CIII ratio and other changes in the apolipoprotein profile. To assess whether diabetic rats were less able to clear triacylglycerol independent of changes in the nature of the lipoproteins, we monitored the clearance of chylomicron-like lipid emulsions in hepatectomized rats. In vivo, emulsion triacylglycerol hydrolysis was not slowed due to diabetes. However, control and diabetic rats, which had been fasted for 16 h, cleared triacylglycerol at about twice the rate of fed rats. Triacylglycerol secretion rates in diabetic and control rats were similar, whether fed or fasted. We conclude that in streptozocin diabetic rats, hypertriglyceridemia was not due to overproduction of chylomicron- or VLDL-triacylglycerol, nor to decreased endothelial lipase activities. Rather, in fed diabetic rats, the triacylglycerol-rich lipoproteins are poorer substrates for lipoprotein lipase. This may lead to slower formation of remnants which would exacerbate slow remnant removal. VLDL of diabetic origin were hydrolysed as efficiently as VLDL from control donors, suggesting that in the fed state the lipolytic defect may be specific for chylomicrons.     

Aquatic nematodes from Ethiopia III

Six species of the genus Eumonhystera, one of which is new to science, are described from bottom samples of Lake Tana and two associated rivers, River Abbay and R. Gelda, Ethiopia. E. geraerti n.sp. can be identified by being the only species in the genus that combines very posteriorly positioned vulva [V% = 68.2 ± 0.7 (67.3–69.3)] with very anteriorly positioned amphids (Amph-ABE/LRW = 0.8). The other five species encountered, E. dispar (Bastian, 1865) Andrássy, 1981, E. filiformis (Bastian, 1865) Andrássy, 1981, E. pseudobulbosa (von Daday, 1896) Andrássy, 1981, E. mwerazii (Meyl, 1957) Andrássy, 1981, and E. vulgaris (de Man, 1880), Andrássy, 1981, are described in detail. The identities of some of the already described populations of E. dispar, E. filiformis and E. vulgaris are questioned and clearer and better ways of defining each of these three species are suggested. A peritrophic-like membrane on the outer side of the brush-border of the intestine of E. mwerazii, a species reported here for the first time out of its type locality, is reported. Also, scanning electronmicrographs of sectioned and entire specimens of E. mwerazii are presented. All species are reported from Ethiopia for the first time.     

A Mycobacterium leprae-specific gene encoding an immunologically recognized 45 kDa protein

By screening a Mycobacterium leprae lambda gt11 expression library with a serum from an Ethiopian lepromatous leprosy (LL) patient a clone was isolated (LL4) belonging to hybridization group III of a panel of previously isolated M. leprae clones. Members of this hybridization group encode a serologically recognized 45 kDa protein. The complete DNA sequences of the partially overlapping clones LL4 and L1 (hybridization group III) are presented and these revealed the presence of an open reading frame (ORF) predicting a protein with a molecular size of 42 448 Da. Southern hybridizations on total genomic DNA of M. Ieprae, Mycobacterium tuberculosis and eight atypical mycobacteria showed that the LL4 DNA fragment is specific for M. Ieprae DNA even under low-stringency conditions. The M. Ieprae specificity of LL4 DNA was further confirmed by the polymerase chain reaction using four different sets of primers. Western blotting analyses showed that the M. Ieprae 45 kDa protein is frequently recognized by antibodies from leprosy patients and that this recognition is specific since no antibodies could be detected in sera of tuberculosis patients. T-cell proliferation assays also demonstrated T-cell recognition by leprosy patients and healthy contacts of the M. Ieprae 45 kDa protein. The specificity of the LL4 DNA region and the 45 kDa antigen that is encoded by hybridization group III could provide unique tools for the development of M. Ieprae-specific immunological and DNA reagents.     

Vaccination with Staphylococcus aureus fibrinogen binding proteins (FgBPs) reduces colonisation of S. aureus in a mouse mastitis model

Abstract A mouse mastitis model was used to study the effect of vaccination with fibrinogen binding proteins and collagen binding protein from Staphylococcus aureus against challenge infection with S. aureus . The mice vaccinated with fibrinogen binding proteins showed reduced rates of mastitis compared with controls. Gross examination of challenged mammary glands of mice showed that the glands of mice immunized with fibronogen binding proteins developed mild intramammary infection or had no pathological changes compared with glands from control mice. Histopathological examination of tissue sections from challenged glands showed that most glands from mice vaccinated with fibrinogen binding protein developed disseminated necrosis or had no pathological changes. A significantly reduced number of bacteria could be recovered in the glands from mice immunized with fibrinogen binding proteins as compared with controls. In a similar study, immunization of mice with collagen binding protein did not induce protection against challenge infection with S. aureus .     

昆虫嗅觉相关蛋白及嗅觉识别机理研究概述

嗅觉是昆虫产生行为的基础之一,在长期进化的过程中昆虫形成了复杂的嗅觉系统,完成这一过程,需要有多种与嗅觉相关的蛋白参与,包括气味结合蛋白、化学感受蛋白、气味受体和感觉神经元膜蛋白等。了解昆虫感受外界信息的嗅觉机制可以帮助我们更好地理解昆虫识别配偶、天敌及寻找食物来源、产卵场地等行为特征,为进一步调控昆虫的行为、防控害虫侵袭、保护和利用有益昆虫奠定基础。本文综述了昆虫嗅觉相关的几类重要蛋白的生化特性和生理功能,并对昆虫气味分子的识别机制、气味分子在昆虫体内运输机制的最新研究进展进行了概述。     

Behavioural thermoregulation and climatic range restriction in the globally threatened Ethiopian Bush‐crow Zavattariornis stresemanni

Climate may influence the distribution and abundance of a species through a number of demographic and ecological processes, but the proximate drivers of such responses are only recently being identified. The Ethiopian Bush‐crow Zavattariornis stresemanni is a starling‐like corvid that is restricted to a small region of southern Ethiopia. It is classified as Endangered in the IUCN Red List of globally threatened species. Previous work suggested that this range restriction is almost perfectly defined by a climate envelope that is cooler than surrounding areas, but the proximate mechanism remains unexplained. The heavily altered habitats which the species inhabits are widespread across Africa, and recent work has shown that the Bush‐crow is behaviourally adaptable and has a catholic diet. We assess whether its enigmatic distribution can be explained by behavioural responses to the higher temperatures that surround its current range. Using environmental niche models and field observations of thermally mediated behaviour, we compare the range restriction and behavioural thermoregulation of the Ethiopian Bush‐crow with those of two sympatric control species that are similar in size and ecology, but have much larger ranges that include hotter environments. White‐crowned Starling Lamprotornis albicapillus and Superb Starling L. superbus occupy similar habitats to the Ethiopian Bush‐crow and all three frequently forage together. We found that the Bush‐crow's range is limited primarily by temperature, with a secondary effect of dry season rainfall, whereas the ranges of the two starling species are better predicted by wet season rainfall alone. Bush‐crows exhibited panting behaviour and moved into the shade of trees at significantly lower ambient temperatures than did the starlings, and their food intake declined more steeply with increasing temperature. These results indicate that the limited geographical range of the Bush‐crow reflects an inability to cope with higher temperatures. This suggests that a species' response to climate change might not be easily predicted by its ecological generalism, and may represent an inherited debt from its evolutionary history.     

Insulin decreases the secretion of apoB-100 from hepatic HepG2 cells but does not decrease the secretion of apoB-48 from intestinal CaCo-2 cells

We compared the acute effect of insulin on the human colonic intestinal epithelial cell line CaCo-2 and the transformed human hepatic cell line HepG2. Over 24 h, 100 nM and 10 µM insulin significantly inhibited the secretion of apolipoprotein (apo) B-100 from HepG2 cells to 63 and 49% of control, respectively. Insulin had no effect on the secretion of apoB-48 from CaCo-2 cells. There was no effect of insulin on the cholesterol ester or free cholesterol concentrations in HepG2 or CaCo-2 cells. HepG2 and CaCo-2 cells bound insulin with high affinity, leading to similar stimulation of insulin receptor protein tyrosine kinase activation. Protein kinase C or mitogen-activated protein kinase activity in the presence or absence of insulin was not correlated with apoB-48 production in CaCo-2 cells. Therefore, insulin acutely decreases the secretion of apoB-100 in hepatic HepG2 cells, but does not acutely modulate the production or secretion of apoB-48 from CaCo-2 intestinal cells.     

Site-directed mutagenesis of an alkaline phytase: influencing specificity, activity and stability in acidic milieu

Site-directed mutagenesis of a thermostable alkaline phytase from Bacillus sp. MD2 was performed with an aim to increase its specific activity and activity and stability in an acidic environment. The mutation sites are distributed on the catalytic surface of the enzyme (P257R, E180N, E229V and S283R) and in the active site (K77R, K179R and E227S). Selection of the residues was based on the idea that acid active phytases are more positively charged around their catalytic surfaces. Thus, a decrease in the content of negatively charged residues or an increase in the positive charges in the catalytic region of an alkaline phytase was assumed to influence the enzyme activity and stability at low pH. Moreover, widening of the substrate-binding pocket is expected to improve the hydrolysis of substrates that are not efficiently hydrolysed by wild type alkaline phytase. Analysis of the phytase variants revealed that E229V and S283R mutants increased the specific activity by about 19% and 13%, respectively. Mutation of the active site residues K77R and K179R led to severe reduction in the specific activity of the enzyme. Analysis of the phytase mutant-phytate complexes revealed increase in hydrogen bonding between the enzyme and the substrate, which might retard the release of the product, resulting in decreased activity. On the other hand, the double mutant (K77R-K179R) phytase showed higher stability at low pH (pH 2.6-3.0). The E227S variant was optimally active at pH 5.5 (in contrast to the wild type enzyme that had an optimum pH of 6) and it exhibited higher stability in acidic condition. This mutant phytase, displayed over 80% of its initial activity after 3 h incubation at pH 2.6 while the wild type phytase retained only about 40% of its original activity. Moreover, the relative activity of this mutant phytase on calcium phytate, sodium pyrophosphate and p-nitro phenyl phosphate was higher than that of the wild type phytase.     

Brood Ball-Mediated Transmission of Microbiome Members in the Dung Beetle,Onthophagus taurus (Coleoptera: Scarabaeidae)

Insects feeding on plant sap, blood, and other nutritionally incomplete diets are typically associated with mutualistic bacteria that supplement missing nutrients. Herbivorous mammal dung contains more than 86% cellulose and lacks amino acids essential for insect development and reproduction. Yet one of the most ecologically necessary and evolutionarily successful groups of beetles, the dung beetles (Scarabaeinae) feeds primarily, or exclusively, on dung. These associations suggest that dung beetles may benefit from mutualistic bacteria that provide nutrients missing from dung. The nesting behaviors of the female parent and the feeding behaviors of the larvae suggest that a microbiome could be vertically transmitted from the parental female to her offspring through the brood ball. Using sterile rearing and a combination of molecular and culture-based techniques, we examine transmission of the microbiome in the bull-headed dung beetle, Onthophagus taurus. Beetles were reared on autoclaved dung and the microbiome was characterized across development. A ~1425 bp region of the 16S rRNA identified Pseudomonadaceae, Enterobacteriaceae, and Comamonadaceae as the most common bacterial families across all life stages and populations, including cultured isolates from the 3^rd instar digestive system. Finer level phylotyping analyses based on lepA and gyrB amplicons of cultured isolates placed the isolates closest to Enterobacter cloacae, Providencia stuartii, Pusillimonas sp., Pedobacter heparinus, and Lysinibacillus sphaericus. Scanning electron micrographs of brood balls constructed from sterile dung reveals secretions and microbes only in the chamber the female prepares for the egg. The use of autoclaved dung for rearing, the presence of microbes in the brood ball and offspring, and identical 16S rRNA sequences in both parent and offspring suggests that the O. taurus female parent transmits specific microbiome members to her offspring through the brood chamber. The transmission of the dung beetle microbiome highlights the maintenance and likely importance of this newly-characterized bacterial community.