Interaction of the Lyme disease spirochete with its tick vector

Borrelia burgdorferi, the causative agent of Lyme disease (along with closely related genospecies), is in the deeply branching spirochete phylum. The bacterium is maintained in nature in an enzootic cycle that involves transmission from a tick vector to a vertebrate host and acquisition from a vertebrate host to a tick vector. During its arthropod sojourn, B. burgdorferi faces a variety of stresses, including nutrient deprivation. Here, we review some of the spirochetal factors that promote persistence, maintenance and dissemination of B. burgdorferi in the tick, and then focus on the utilization of available carbohydrates as well as the exquisite regulatory systems invoked to adapt to the austere environment between blood meals and to signal species transitions as the bacteria traverse their enzootic cycle. The spirochetes shift their source of carbon and energy from glucose in the vertebrate to glycerol in the tick. Regulation of survival under limiting nutrients requires the classic stringent response in which Rel_Bbu controls the levels of the alarmones guanosine tetraphosphate and guanosine pentaphosphate (collectively termed (p)ppGpp), while regulation at the tick–vertebrate interface as well as regulation of protective responses to the blood meal require the two‐component system Hk1/Rrp1 to activate production of the second messenger cyclic‐dimeric‐GMP (c‐di‐GMP).     

The Borrelia burgdorferi circular plasmid cp26: conservation of plasmid structure and targeted inactivation of the ospC gene

The 26 to 28 kb circular plasmid of B. burgdorferi sensu lato (cp26) is ubiquitous among bacteria of this group and contains loci implicated in the mouse–tick transmission cycle. Restriction mapping and Southern hybridization indicated that the structure of cp26 is conserved among isolates from different origins and culture passage histories. The cp26 ospC gene encodes an outer surface protein whose synthesis within infected ticks increases when the ticks feed, and whose synthesis in culture increases after a temperature upshift. Previous studies of ospC coding sequences showed them to have stretches of sequence apparently derived from the ospC genes of distantly related isolates by homologous recombination after DNA transfer. We found conservation of the promoter regions of the ospC and guaA genes, which are divergently transcribed. We also demonstrated that the increase in OspC protein after a temperature upshift parallels increases in mRNA levels, as expected if regulatory regions adjoin the conserved sequences in the promoter regions. Finally, we used directed insertion to inactivate the ospC gene of a non-infectious isolate. This first example of directed gene inactivation in B. burgdorferi shows that the OspC protein is not required for stable maintenance of cp26 or growth in culture.     

A novel rearrangement of occludin causes brain calcification and renal dysfunction

Pediatric intracranial calcification may be caused by inherited or acquired factors. We describe the identification of a novel rearrangement in which a downstream pseudogene translocates into exon 9 of OCLN, resulting in band-like brain calcification and advanced chronic kidney disease in early childhood. SNP genotyping and read-depth variation from whole exome sequencing initially pointed to a mutation in the OCLN gene. The high degree of identity between OCLN and two pseudogenes required a combination of multiplex ligation-dependent probe amplification, PCR, and Sanger sequencing to identify the genomic rearrangement that was the underlying genetic cause of the disease. Mutations in exon 3, or at the 5–6 intron splice site, of OCLN have been reported to cause brain calcification and polymicrogyria with no evidence of extra-cranial phenotypes. Of the OCLN splice variants described, all make use of exon 9, while OCLN variants that use exons 3, 5, and 6 are tissue specific. The genetic rearrangement we identified in exon 9 provides a plausible explanation for the expanded clinical phenotype observed in our individuals. Furthermore, the lack of polymicrogyria associated with the rearrangement of OCLN in our patients extends the range of cranial defects that can be observed due to OCLN mutations.     

甲亢甲低患者肝纤维化指标检测的临床意义分析

目的:探讨甲状腺功能亢进(甲亢)、甲状腺功能减低(甲低)与肝纤维化指标的关系及其可能的机制。方法:采用放射免疫分析法(R1A)检测57例甲亢患者、43例甲低患者、39例甲亢治疗后甲状腺激素正常者和50例健康成人的血清Ⅳ型胶原(1V-C)、Ⅲ型前胶原(PCⅢ)、透明质酸(HA)、层粘连蛋白(LN)、T3、T4、FT3、FT4、TSH、TGA、TMA含量。结果:甲亢患者组血清中1V-C、PCⅢ含量比正常对照组及甲低患者组显著性增高(P<0.05);治疗后甲状腺激素下降,1V-C、PCⅢ含量也随之下降(与治疗前比较P<0.01);HA、LN在四组中无显著性差异(P>0.05),在甲亢治疗前后无显著性差异(P>0.05)。各项肝纤维化指标与TGA、TMA的阳性率无关。结论:甲亢患者可有不同程度的肝功能损害,血清中甲状腺激素水平增高时,1V-C、PCⅢ水平也增高,在用1V-C、PCⅢ判断肝纤维化时应注意有无甲状腺疾病特别是甲亢。未发现HA、LN含量与甲状腺激素水平的关系。     

脂肪间充质干细胞来源外泌体的功能

外泌体是细胞外膜质纳米囊泡,将蛋白质、核酸(DNA和RNA)转运到靶细胞中,介导局部和系统的细胞间通信,从而改变受体细胞的行为.这些小泡在许多生物功能中发挥重要作用,如脂肪合成、免疫调节、神经再生和肿瘤调节等.脂肪间充质干细胞目前被认为是细胞治疗和再生医学领域中一种功能丰富的工具,可产生和分泌多种外泌体,继承细胞的多种...     

Modulation of intestinal protein synthesis and protease mRNA by luminal and systemic nutrients

Route of nutrient supply is important in regulation of intestinal protein metabolism, because total parenteral nutrition, compared with enteral feeding, leads to profound atrophy. Participation of the fractional rate of protein synthesis (Ks), their degradation in regulation of gut protein balance, and their possible modulation by specific nutrients are the focus of our work. We developed an in situ experimental system that allows controlled exposure of intestinal mucosa to nutrients systemically, luminally, or both. We examined the effects of systemic glucose and amino acid (AA) infusion in overnight-fasted piglets. Jejunal segments within each piglet were simultaneously, luminally perfused with solutions containing various AAs or glucose. Intravenous infusion of glucose increased mucosal Ks by 16% (P < 0.05), whereas intravenous infusion of AA had no effect on Ks. Systemic glucose infusion had no effect on mRNA levels for components of the ubiquitin-proteasome proteolytic pathway. However, levels of these mRNA were reduced by intravenous or luminal AA supply. This effect was greatest (-50%) when highest tissue concentrations of AAs were achieved by the simultaneous infusion of AA by both routes (P < 0.05). Our findings suggest that not only is the modulation of protein balance in the intestine in response to nutrients in part attributable to anabolic stimulation of protein synthesis initiated by the systemic appearance of glucose, but a fall in protein degradation is also a likely contributor. AAs appear to be a key factor required to reduce expression of genes connected with proteolysis.     

羊草草地生长季放牧山羊采食量和食性选择

在松嫩平原羊草草地,通过控制放牧实验对山羊的时限采食量和食性选择进行了研究。结果表明：（1）5－9月份,山羊的时限（1h）采食量平均为0.42kg干物质,其季节动态为5月份最低,随季节推移不断增大,8月份达到最大,9月份又有所减小;时限采食量基本上随放牧率减小而增大,但在最低放牧率小区有所减小。（2）山羊的食性选择随季节推移和放牧率不同而变化。（3）山羊对20－25cm高度草层的选择性最高;各高度草层的食性选择指数随季节推移和放牧率不同而变化;山羊对不同植物的高度选择性存在差异,但高度选择指数的最大值都在15－30cm范围内。     

Cellular machinery of wood production: differentiation of secondary xylem in Pinus contorta var. latifolia

The objectives of this study were to define cell structure during pine secondary xylem development and to integrate this information with current knowledge of the biochemistry and physiology of secondary cell wall biosynthesis in gymnosperms. Lodgepole pine (Pinus contorta var. latifolia Englem.) cambium and secondary xylem were cryofixed using high pressure freezing and freeze-substitution which allowed excellent preservation of the cell structure of developing secondary xylem and enabled high-resolution transmission electron microscopic viewing of these cells for the first time. In contrast to their precursors in the adjacent cambial zone, developing tracheids were active in secondary wall deposition, with abundant cortical microtubules and developing bordered pits. These cells were also characterized by unusual Golgi structures: the trans-Golgi network was highly developed and the associated vesicles were large and darkly stained. These unusual Golgi structures persisted throughout the period of xylem maturation until programmed cell death occurred. Immuno-cytochemistry and enzyme-gold probes were used to investigate the distribution of key secretory products (mannans) and a lignification-associated enzyme (coniferin beta-glucosidase) during xylogenesis. Mannans were localized to the secondary cell wall, the trans-Golgi cisternae and trans-Golgi network vesicles of developing xylem. Coniferin beta-glucosidase was found only in the secondary cell wall. The cell wall localization of coniferin beta-glucosidase, the enzyme responsible for cleaving glucose from coniferin to generate free coniferyl alcohol, provides a mechanism to de-glucosylate monolignols in muro. A two-step model of lignification of conifer tracheids is proposed. First, Golgi-mediated secretion deposits monolignols into the cell wall, where they polymerize in cell corners and middle lamella. Secondly, cell lysis releases stored, vacuolar monolignol glucosides into the wall where they are deglucosylated and their polymerization is influenced by the wall environment including the lignin deposited earlier.     

Hydrolytic enzymes of leaf-cutting ant fungi

The production of enzymes and the colonization of leaves by Leucoagaricus gongylophorus were investigated to further understand the digestive interactions of leaf-cutting ant colonies. The enzymes detected were indicative of a saprophytic origin of this fungus, producing all the enzymes necessary for plant tissue breakdown. Enhanced activities of certain enzymes in the fungus garden extracts may be due to the particular behaviour of the adult worker ants that concentrate fungal acquired enzymes in the rectal fluid and subsequently defaecate these enzymes onto the leaves. The production of chitinases by the fungus may be an ancestral vestige of lower attines, and may have a role as agonists of invading microbes. Growth of the fungus on plant cell wall medium resulted in highest enzyme activity against pectin, reflecting the fact that polygalacturonans comprise the main matrix of the primary plant cell wall. SEM shows that L. gongylophorus does not form specialized structures for cell wall penetration, but gains access to the inner plant tissue at the cut edges of the leaf fragments. Enzymes secreted by the fungus were compared to those seen in larval and adult leaf-cutting ants, demonstrating the inter-dependence of the symbiotic relationship between the ants and their fungi.     

Forelimb indicators of prey‐size preference in the Felidae

The forelimbs, along with the crania, are an essential part of the prey‐killing apparatus in cats. Linear morphometrics of the forelimbs were used to determine the morphological differences between felids that specialize on large prey, small prey, or mixed prey. We also compared the scaling of felid forelimbs to those of canids to test whether prey capture strategies affect forelimb scaling. Results suggest that large prey specialists have relatively robust forelimbs when compared with smaller prey specialists. This includes relatively more robust humeri and radii, relatively larger distal ends of the humerus, and relatively larger articular areas of the humerus and radius. Large prey specialists also had relatively longer olecranon processes of the ulna and wider proximal paws. These characters are all important for subduing large prey while the cat positions itself for the killing bite. Small prey specialists have relatively longer distal limb elements for swift prey capture, and mixed prey specialists had intermediate values with relatively more robust metacarpals. Arboreal felids also had more robust limbs. They had relatively longer proximal phalanges for better grip while climbing, and a relatively short brachial index (radius to humerus ratio). Additionally, we found that felids and canids differ in forelimb scaling, which emphasizes the dual use of forelimbs for locomotion and prey capture in felids. This morphometric technique worked well to separate prey‐size preference in felids, but did not work as well to separate locomotor groups, as scansorial and terrestrial felids were not clearly distinguished. J. Morphol. 2009. © 2009 Wiley‐Liss, Inc.