The Hemagglutinin of Bat-Associated Influenza Viruses Is Activated by TMPRSS2 for pH-Dependent Entry into Bat but Not Human Cells

New World bats have recently been discovered to harbor influenza A virus (FLUAV)-related viruses, termed bat-associated influenza A-like viruses (batFLUAV). The internal proteins of batFLUAV are functional in mammalian cells. In contrast, no biological functionality could be demonstrated for the surface proteins, hemagglutinin (HA)-like (HAL) and neuraminidase (NA)-like (NAL), and these proteins need to be replaced by their human counterparts to allow spread of batFLUAV in human cells. Here, we employed rhabdoviral vectors to study the role of HAL and NAL in viral entry. Vectors pseudotyped with batFLUAV-HAL and -NAL were able to enter bat cells but not cells from other mammalian species. Host cell entry was mediated by HAL and was dependent on prior proteolytic activation of HAL and endosomal low pH. In contrast, sialic acids were dispensable for HAL-driven entry. Finally, the type II transmembrane serine protease TMPRSS2 was able to activate HAL for cell entry indicating that batFLUAV can utilize human proteases for HAL activation. Collectively, these results identify viral and cellular factors governing host cell entry driven by batFLUAV surface proteins. They suggest that the absence of a functional receptor precludes entry of batFLUAV into human cells while other prerequisites for entry, HAL activation and protonation, are met in target cells of human origin.     

Calcified gallstone in a 3 year-old boy: a case report

ABSTRACT: BACKGROUND: Gallstones are relatively rare in children. At-risk populations include patients suffering from hemolysis syndromes. Regardless of etiology, these patients usually will present with postprandial abdominal pain, and ultrasonography is the mainstay of diagnosis. However, some gallstones are radiopaque and can be visualized on plain abdominal radiography. CASE PRESENTATION: We present the uncommon but classic plain x-ray finding of a calcified gallstone in a 3 year-old Hispanic boy. He was treated with elective laparoscopic cholecystectomy. CONCLUSIONS: Cholelithiasis is rare in children, and calcified stones that will appear on plain abdominal x-rays are even rarer. If symptomatic, cholecystectomy by a pediatric surgeon is the treatment of choice. We discuss some of the recent developments in treatment of this condition in this patient population.     

Mariculture in Israel – past achievements and future directions in raising rotifers as food for marine fish larvae

Marine fish production is now being carried out afteralmost two decades of research. The production ofseabream (Sparus aurata), which reached over 750tons in 1995, is expected to reach an annualproduction ranging between 4000 - 12 700 metric tonsby year 2010. The anticipated introduction of newspecies and its expansion to the Mediterranean shoreline will help in leading the increased maricultureproduction. Two marine fish hatcheries that operatetoday in Israel produce 7 million fingerlings a year.Traditionally, aquaculture in Israel raises fish ininland freshwater ponds and irrigation reservoirs. Inaddition, Lake Kinneret, the only freshwater lake inIsrael, is stocked yearly with juvenile fish raised inlocal hatcheries (tilapia) or imported fromMediterranean countries (mugil). While culture offreshwater teleost species (carp) was introduced morethan fifty years ago, mariculture started on acommercial scale less than 5 years ago. The limitedsupply of freshwater will accelerate the futureculture of marine species.The bottleneck of almost all marine finfish productionlies in obtaining adequate numbers of fingerlings, dueto their high mortality at early life stages. Theproduction is hindered by inadequate supply of food toearly larval stages which require live food.Development of technologies in Israel for masscultivation of food chain organisms including algae,rotifers and brine shrimp followed their developmentin other parts of the world, most notably thoseachieved in Japan. The local commercial scaleproduction of rotifers relies on several batch orsemi-continuous cultures in conical or flatbottomrectangular containers that supply daily 0.6-4billion rotifers in each hatchery. Originally arelatively large local Brachionus plicatilisstrain was used, but later smaller B.rotundiformis strains were introduced, resulting ina mixture of undefined strains. The incorporation ofalgae (Nannochloropsis sp.) generated in highyield raceways contributes to the reliability ofrotifer cultures. Algae are supplied directly from theraceways or centrifuged and stored as a frozen pasteuntil required in the hatchery. The current dependablesupply of live cultures reduces the need for preservedstocks of rotifers, either as resting eggs or keptalive at low temperatures. To the fish grower,rotifers are live food capsules that deliver essentialnutrients (e.g. long chain unsaturated fatty acids)for growth and survival of fish larvae. Research aimedat replacing live food with chemically definedmicrodiets could reveal physiological principles inprey recognition and digestion of food by marine fishlarvae.     

Differential and gonad stage-dependent roles of kisspeptin1 and kisspeptin2 in reproduction in the modern teleosts, morone species

Kisspeptin is an important regulator of reproduction in many vertebrates. The involvement of the two kisspeptins, Kiss1 and Kiss2, and their receptors, Gpr54-1 and Gpr54-2, in controlling reproduction was studied in the brains of the modern teleosts, striped and hybrid basses. In situ hybridization and laser capture microdissection followed by quantitative RT (QRT)-PCR detected coexpression of kiss1 and kiss2 in the hypothalamic nucleus of the lateral recess. Neurons expressing gpr54-1 and gpr54-2 were detected in several brain regions. In the preoptic area, gpr54-2 was colocalized in GnRH1 neurons while gpr54-1 was expressed in cells attached to GnRH1 fibers, indicating two different modes of GnRH1 regulation. The expression of all four genes was measured in the brains of males and females at different life stages using QRT-PCR. The levels of kiss1 and gpr54-1 mRNA, the latter being expressed in minute levels, were consistently lower than those of kiss2 and gpr54-2. While neither gene's expression increased at prepuberty, all were dramatically elevated in mature females. The levels of kiss2 mRNA increased also in mature males. Kiss1 peptide was less potent than Kiss2 in elevating plasma luteinizing hormone levels and in up-regulating gnrh1 and gpr54-2 expression in prepubertal hybrid bass in vivo. In contrast, during recrudescence, Kiss1 was more potent than Kiss2 in inducing luteinizing hormone release, and Kiss2 down-regulated gnrh1 and gpr54-2 expression. This is the first report in fish to demonstrate the alternating actions and the importance of both neuropeptides for reproduction. The organization of the kisspeptin system suggests a transitional evolutionary state between early to late evolving vertebrates.     

Preliminary in vitro study on the effect of xanthohumol on rumen methanogenesis

There is a great interest in reducing the methane emission from ruminants as one possible cause of global warming. The aim of the presented study was to determine the effects of xanthohumol, one of the hop (Humulus lupulus) phytofactors, on methane production, microbial population and basic parameters of ruminal fermentation. The experiment was carried out in a batch culture system. The basic substrate (400 mg) consisting of meadow hay and barley meal (60:40) was supplemented with 0 (Control), 0.1, 0.2 or 1.0 mg of xanthohumol. The basic parameters of rumen fermentation and composition of microbial population were measured after 24 h of incubation. For the first time, the results of this in vitro study have demonstrated that xanthohumol is capable to reduce the methane production, even at the lowest dose applied (0.1 mg/400 mg). The observed reduction in methane production by 12-13% was not accompanied by altering the basic rumen fermentation parameters. However, the practical utility of this supplement needs further investigation under long-term in vivo conditions.     

TMPRSS2 Isoform 1 Activates Respiratory Viruses and Is Expressed in Viral Target Cells

The cellular protease TMPRSS2 cleaves and activates the influenza virus hemagglutinin (HA) and TMPRSS2 expression is essential for viral spread and pathogenesis in mice. Moreover, severe acute respiratory syndrome coronavirus (SARS-CoV) and other respiratory viruses are activated by TMPRSS2. However, previous studies on viral activation by TMPRSS2 focused on a 492 amino acids comprising form of the protein (isoform 2) while other TMPRSS2 isoforms, generated upon alternative splicing of the tmprss2 mRNA, have not been characterized. Here, we show that the mRNA encoding a TMPRSS2 isoform with an extended N-terminal cytoplasmic domain (isoform 1) is expressed in lung-derived cell lines and tissues. Moreover, we demonstrate that TMPRSS2 isoform 1 colocalizes with HA and cleaves and activates HA. Finally, we show that isoform 1 activates the SARS-CoV spike protein for cathepsin L-independent entry into target cells. Our results indicate that TMPRSS2 isoform 1 is expressed in viral target cells and might contribute to viral activation in the host.     

Biotechnology and aquaculture of rotifers

Biotechnology can be defined as any technology that involves living organisms or their derivatives. In applying this definition to rotifers, we focus on their contribution in culturing of early larval stages of marine fish. After almost four decades of marine fish culture in captivity, the success of this worldwide industry is still quite dependent on mass culture of the species Brachionus plicatilis and B. rotundiformis. In mass culture, the rotifers are continuously driven to reproduce at high rates, in relatively extreme environmental conditions of high population density and high loads of organic matter. Therefore, the success of mass cultures and future improvements in these systems relies on a close interaction between basic and applied studies of rotifers. In the present review, we will attempt to analyze why rotifers are suitable for early life stages of fish and to describe, in general, methodologies that have been devised for reliable supply of rotifers in large quantities. Problems associated with rotifer production, nutritional quality and effect on fish health and nutrition, will be discussed. Research on B. plicatilis and B. rotundiformis has increased enormously during the past three decades and these two species are the best-studied rotifers so far. While much of the research on these species is directed or devoted to the needs of aquaculture industry, they are also used as models for addressing basic biological questions, due to the relative ease of culture and their availability. Studies on feeding, pheromones, speciation in rotifers, the occurrence and putative hormones involved in sexual and asexual reproduction and production of resting eggs, are few examples of such studies. Rotifers will probably maintain their role as food organism for fish larvae, in spite of attempts to replace them with more accessible formulated food. Development of new culture methods that will improve the nutritional quality and production efficiency of rotifers may result in more diversified and flexible tasks for these organisms in aquaculture.