Crustacean hyperglycemic hormone (CHH) neuropeptidesfamily: Functions, titer, and binding to target tissues.

The removal of the eyestalk (s) induces molting and reproduction promoted the presence of regulatory substances in the eyestalk (ES), particularly medulla terminalis X-organ and the sinus gland (MTXO-SG). The PCR-based cloning strategies have allowed for isolating a great number of cDNAs sequences of crustacean hyperglycemic hormone (CHH) neuropeptides family from the eyestalk and non-eyestalk tissues, e.g., pericardial organs and fore- and hindguts. However, the translated corresponding neuropeptides in these tissues, their circulating concentrations, the mode of actions, and specific physiological functions have not been well described. The profiles of CHH neuropeptides present in the MTXO-SG may differ among decapod crustacean species, but they can be largely divided into two sub-groups on the basis of structural homology: (1) CHH and (2) molt-inhibiting hormone (MIH)/mandibular organ-inhibiting hormone (MOIH)/vitellogenesis/gonad-inhibiting hormone (V/GIH). CHH typically elevating the level of circulating glucose from animals under stressful conditions (hyper- and hypothermia, hypoxia, and low salinity) has multiple target tissues and functions such as ecdysteroidogenesis, osmoregulation, and vitellogenesis. Recently, MIH, known for exclusively suppressing ecdysteroidogenesis in Y-organs, is also reported to have an additional role in vitellogenesis of adult female crustacean species, suggesting that some CHH neuropeptides may acquire an extra regulatory role in reproduction at adult stage. This paper reviews the regulatory roles of CHH and MIH at the levels of specific functions, temporal and spatial expression, titers, their binding sites on the target tissues, and second messengers from two crab species: the blue crab, Callinectes sapidus, and the European green crab, Carcinus maenas. It further discusses the diverse regulatory roles of these neuropeptides and the functional plasticity of these neuropeptides in regard to life stage and species-specific physiology.

Global-scale analyses of chemical ecology and population genetics in the invasive Argentine ant.

Ants are some of the most abundant and ecologically successful terrestrial organisms, and invasive ants rank among the most damaging invasive species. The Argentine ant is a particularly well-studied invader, in part because of the extreme social structure of introduced populations, known as unicoloniality. Unicolonial ants form geographically vast supercolonies, within which territorial behaviour and intraspecific aggression are absent. Because the extreme social structure of introduced populations arises from the widespread acceptance of conspecifics, understanding how this colonymate recognition occurs is key to explaining their success as invaders. Here, we present analyses of Argentine ant recognition cues (cuticular hydrocarbons) and population genetic characteristics from 25 sites across four continents and the Hawaiian Islands. By examining both hydrocarbon profiles and microsatellite genotypes in the same individual ants, we show that native and introduced populations differ in several respects. Both individual workers and groups of nestmates in the introduced range possess less diverse chemical profiles than ants in the native range. As previous studies have reported, we also find that introduced populations possess much lower levels of genetic diversity than populations in the native range. Interestingly, the largest supercolonies on several continents are strikingly similar to each other, suggesting that they arose from a shared introduction pathway. This high similarity suggests that these geographically far-flung ants may still recognize and accept each other as colonymates, thus representing distant nodes of a single, widely distributed supercolony. These findings shed light on the behaviour and sociality of these unicolonial invaders, and pose new questions about the history and origins of introduced populations.

Genetics and behavior of a colonizing species: the invasive Argentine ant.

Baker and Stebbins's 1965 book The Genetics of Colonizing Species aimed to draw together scientists from a variety of disciplines to provide a conceptual framework for the study of species introductions. A goal of their volume was to examine how studies on biological invasions could be used to provide insight into basic research questions as well as to develop practical strategies for control. In this article, we attempt to follow the goals of Baker and Stebbins by reviewing work on the genetics and behavior of a widespread colonizing species, the Argentine ant (Linepithema humile). Specifically, we examine the evolutionary changes that have taken place as a result of this species being introduced into new environments and synthesize recent research on Argentine ants from the perspective of population genetics, recognition systems, and the mechanisms that may underlie their ecological success.

B(0)-B(0) mixing in unquenched lattice QCD.

We present an unquenched lattice calculation for the B(0)-B(0) transition amplitude. The calculation, carried out at an inverse lattice spacing 1/a=2.22(4) GeV, incorporates two flavors of dynamical quarks described by the O(a)-improved Wilson fermion action and heavy quarks described by nonrelativistic QCD. Particular attention is paid to the uncertainty that arises from the chiral extrapolation, especially the effect of pion loops, for light quarks, which we find could be sizable for the leptonic decay constant, whereas it is small for the B parameters. We obtain f(B(d))=191(10)(+12-22) MeV, f(B(s))/f(B(d))=1.13(3)(+13-2), B(B(d))(m(b))=0.836(27)(+56-62), B(B(s))/B(B(d))=1.017(16)(+56-17), and xi=1.14(3)(+13-2), where the first error is statistical, and the second is systematic, including uncertainties due to chiral extrapolation, finite lattice spacing, heavy quark expansion, and perturbative operator matching.

Relationships among native and introduced populations of the Argentine ant (Linepithema humile) and the source of introduced populations.

The Argentine ant (Linepithema humile) is a damaging invasive species that has become established in many Mediterranean-type ecosystems worldwide. To identify likely sources of introduced populations we examined the relationships among native Linepithema populations from Argentina and Brazil and introduced populations of L. humile using mitochondrial cytochrome b sequence data and nuclear microsatellite allele frequencies. The mitochondrial phylogeny revealed that the populations in Brazil were only distantly related to both the introduced populations and the native populations in Argentina, and confirmed that populations in Brazil, previously identified as L. humile, are likely a different species. The microsatellite-based analysis provided resolution among native and introduced populations of L. humile that could not be resolved using the mitochondrial sequences. In the native range, colonies that were geographically close to one another tended to be genetically similar, whereas more distant colonies were genetically different. Most samples from the introduced range were genetically similar, although some exceptions were noted. Most introduced populations were similar to native populations from the southern Rio Parana and were particularly similar to a population from Rosario, Argentina. These findings implicate populations from the southern Rio Parana as the most likely source of introduced populations. Moreover, these data suggest that current efforts to identify natural enemies of the Argentine ant for biological control should focus on native populations in the southern Rio Parana watershed.

Purification and characterization of a beta-glucuronidase from Aspergillus niger.

A beta-glucuronidase from Pectinex Ultra SP-L, a commercial pectolytic enzyme preparation from Aspergillus niger, was purified 170-fold by ion-exchange chromatography and gel filtration. Apparent M(r) of the purified enzyme, estimated by denaturing gel electrophoresis and size-exclusion chromatography, were 68,000 and 71,000, respectively, indicating that the enzyme is a monomeric protein. It released uronic acids not only from p-nitrophenyl beta-glucosiduronic acid (PNP-GlcA) but also from acidic galactooligosaccharides carrying either beta-D-glucosyluronic or 4-O-methyl-beta-D-glucosyluronic residues at the nonreducing termini through beta-(1-->6)-glycosidic linkages. The enzyme exhibited a maximal activity toward these substrates at pH 3.0. A regioisomer, 3-O-beta-glucosyluronic acid-galactose, was unsusceptible to the enzyme. The enzyme did act on a polymer substrate, releasing uronic acid from the carbohydrate portion of a radish arabinogalactan-protein modified by treatment with fungal alpha-L-arabinofuranosidase. The enzyme produced acidic oligosaccharides by transglycosylation, catalyzing the transfer of uronic acid residues of PNP-GlcA and 6-O-beta-glucosyluronic acid-galactose to certain exogenous acceptor sugars such as Gal, N-acetylgalactosamine, Glc, and xylose.

Population genetics and colony structure of the Argentine ant (Linepithema humile) in its native and introduced ranges.

Introduced species often possess low levels of genetic diversity relative to source populations as a consequence of the small population sizes associated with founder events. Additionally, native and introduced populations of the same species can possess divergent genetic structuring at both large and small geographic scales. Thus, genetic systems that have evolved in the context of high diversity may function quite differently in genetically homogeneous introduced populations. Here we conduct a genetic analysis of native and introduced populations of the Argentine ant (Linepithema humile) in which we show that the population-level changes that have occurred during introduction have produced marked changes in the social structure of this species. Native populations of the Argentine ant are characterized by a pattern of genetic isolation by distance, whereas this pattern is absent in introduced populations. These differences appear to arise both from the effects of recent range expansion in the introduced range as well as from differences in gene flow within each range. Relatedness within nests and colonies is lower in the introduced range than in the native range as a consequence of the widespread genetic similarity that typifies introduced populations. In contrast, nestmates and colony-mates in the native range are more closely related, and local genetic differentiation is evident. Our results shed light on the problem posed for kin selection theory by the low levels of relatedness that are characteristic of many unicolonial species and suggest that the loss of genetic variation may be a common mechanism for the transition to a unicolonial colony structure.

A new endo aortic occlusion balloon for limited access cardiac surgery: development and clinical evaluation.

In recent years, minimally invasive cardiac surgery (MICS), or limited access cardiac surgery, has been presented as a promising operative procedure. We developed a new balloon device that is inserted directly into the ascending aorta to stop the heart during limited access cardiac surgery. The balloon has a three lumen structure: balloon lumen port, cardioplegia/vent lumen port, and aortic root lumen port. This direct EAC balloon catheter, designed to be inserted directly into the ascending aorta, is different from the Heartport system. The Heartport EAC balloon catheter is inserted into the aorta via an artery in the lower limb, making lower limb arterial disease a key concern. Our Direct Endo Aortic Clamp (EAC) balloon overcomes this problem. The device was clinically used in seven cardiac cases. All patients were discharged within 5 postoperative days, confirming the utility of the device.

Determination of amino acid sequence and site of mRNA expression of four vitellins in the giant freshwater prawn, Macrobrachium rosenbergii.

Four major yolk proteins, designated as vitellins (Vns) Macr-VnA, B, C, and D, were extracted from mature ovaries of Macrobrachium rosenbergii. These were purified to homogeneity by reversed-phase high performance liquid chromatography (HPLC) employing a unique separation system based on the hydrophobic properties of the Vn molecule. Using standard techniques of protein sequencing, more than 33 N-terminal and 57 internal amino acid residues were determined for each of the four Vns. The cDNA fragments encoding the four Vns were amplified by PCR using degenerate oligonucleotide primers derived from the N-terminal and internal amino acid sequences. These cDNA fragments were cloned, sequenced, and used as probes to examine the transcription of mRNAs encoding the four Vns. Significant accumulations of these mRNAs were observed in female hepatopancreas only, while mRNA expression was not detected in male hepatopancreas or any other female tissue including ovary, subepidermal adipose tissue, gill, and muscle. This is the first occasion in Crustacea in which multiple Vns were demonstrated to be synthesized simultaneously in a single tissue.

Treatment of acute stanford type B aortic dissection with a novel cylindrical balloon catheter in dogs.

BACKGROUND: Despite recent progress in medical and surgical treatment, acute type B aortic dissection still carries a high mortality rate. We have developed a novel cylindrical balloon catheter for less invasive treatment to block the entry of the dissection and induce thrombotic occlusion of the false lumen. The balloon has the shape of a sheet when deflated but a double-cylinder shape when inflated. Therefore, aortic blood flow is maintained through the cylindrical lumen during balloon inflation. METHODS AND RESULTS: Six beagle dogs underwent a left thoracotomy at the 6th intercostal space. An acute dissection of 4-cm length was created surgically on the descending aorta. The balloon catheter was inserted through the distal descending aorta and advanced to the entry site. The balloon catheter was inflated for 6 hours. The blood flow in the descending aorta and the position of the balloon was monitored by color Doppler echovasculography. Four dogs were killed humanely on the following day and 2 dogs 10 days after the surgery. The descending aorta was examined macroscopically and microscopically in all dogs. In all dogs, the false lumen was occluded by thrombi. Although no dog had clinical evidence of distal thromboembolism, 2 of the 4 dogs that were killed on the second postoperative day had fresh mural thrombi in the true lumen. CONCLUSIONS: The false lumen of the acute type B aortic dissection was effectively occluded by the novel cylindrical balloon catheter in the canine experimental model. The thrombus formation in the true lumen is the problem to be solved.

A novel action of the proton pump inhibitor rabeprazole and its thioether derivative against the motility of Helicobacter pylori.

The motility of Helicobacter pylori was maximum at 37 degrees C and at pH 6. A newly developed proton pump inhibitor, rabeprazole (RPZ), and its thioether derivative (RPZ-TH) markedly inhibited the motility of H. pylori. The concentrations of the drug necessary to inhibit 50% of the motility were 0.25, 16, 16, and >64 microgram/ml for RPZ-TH, RPZ, lansoprazole, and omeprazole, respectively. No such inhibitory effects were observed with H(2) blockers or anti-H. pylori agents. The motilities of Campylobacter jejuni and C. coli-but not those of Vibrio cholerae O1 and O139, Vibrio parahaemolyticus, Salmonella enterica serovar Typhimurium, and Proteus mirabilis-were also inhibited. Prolonged incubation with RPZ or RPZ-TH inhibited bacterial growth of only H. pylori, except for a turbid colony mutant. The results indicate that RPZ and RPZ-TH have a characteristic inhibitory effect against the motility of H. pylori (spiral-shaped bacteria), which is distinguished from that against bacterial growth.

Single low-dose administration of human recombinant hepatocyte growth factor attenuates intimal hyperplasia in a balloon-injured rabbit iliac artery model.

BACKGROUND: Previous studies have shown that repeated systemic administration of human recombinant hepatocyte growth factor (hrHGF) in mg/kg levels modulates the wound-healing process in various diseases. Recently, HGF has been characterized as one of the most potent endothelial-cell-specific growth factors. We tested our hypothesis that local delivery of hrHGF, even at low microg/kg levels (> or =2 orders of magnitude lower than systemically administered doses), might attenuate neointimal hyperplasia in response to vascular injury via accelerated reendothelialization. METHODS AND RESULTS: The iliac artery was denuded in 16 New Zealand White rabbits (3 kg), followed by administration, via a drug delivery catheter, of either hrHGF (10 microg; n = 11) or control vehicle (n=5) over 20 minutes. In pilot studies using this device, the drug permeated into the medial tissues, where it persisted for > or =24 hours. Four weeks after the local delivery of hrHGF, computer-assisted morphometric analysis revealed significant reduction in the intimal area (hrHGF, 0.37+/-0.21 versus control, 0.68+/-0.16 mm(2), mean +/- SD; P<0.05) but no change in the medial area (hrHGF, 1.03+/-0.21 versus control, 1.10+/-0.52 mm(2)). Scanning electron microscopy revealed extensive endothelialization with regular and confluent endothelial cell layer regeneration in the hrHGF-treated vessels. CONCLUSIONS: Accelerated endothelialization after local delivery of hrHGF, a novel and potent endothelial cell mitogen, effectively attenuates neointimal proliferation even after single low-dose administration. This observation could have potential therapeutic implications in the prevention of restenosis after angioplasty.

Reduced genetic variation and the success of an invasive species.

Despite the severe ecological and economic damage caused by introduced species, factors that allow invaders to become successful often remain elusive. Of invasive taxa, ants are among the most widespread and harmful. Highly invasive ants are often unicolonial, forming supercolonies in which workers and queens mix freely among physically separate nests. By reducing costs associated with territoriality, unicolonial species can attain high worker densities, allowing them to achieve interspecific dominance. Here we examine the behavior and population genetics of the invasive Argentine ant (Linepithema humile) in its native and introduced ranges, and we provide a mechanism to explain its success as an invader. Using microsatellite markers, we show that a population bottleneck has reduced the genetic diversity of introduced populations. This loss is associated with reduced intraspecific aggression among spatially separate nests, and leads to the formation of interspecifically dominant supercolonies. In contrast, native populations are more genetically variable and exhibit pronounced intraspecific aggression. Although reductions in genetic diversity are generally considered detrimental, these findings provide an example of how a genetic bottleneck can lead to widespread ecological success. In addition, these results provide insights into the origin and evolution of unicoloniality, which is often considered a challenge to kin selection theory.

New multifunctional percutaneous transluminal coronary angioplasty catheter device capable of balloon inflation, local drug delivery and coronary perfusion.

A new percutaneous transluminal coronary angioplasty catheter with multiple functions of balloon inflation, local drug delivery and coronary perfusion has been devised. The device consists of an inflatable lumen, a drug delivery lumen, and a perfusion (or guide wire) lumen. A drug can be infused from the port located distal to the inflated balloon during continuous blood perfusion via the perfusion lumen. Fluorescence-labeled heparin and peroxidase administered using the device permeated into denuded vessel tissues during ongoing perfusion and remained there for over 24 hr. This prototype device indicates the potential therapeutic implications of the concepts of the device.

Molecular Characterization of a cDNA Encoding Vitellogenin and Its Expression in the Hepatopancreas and Ovary during Vitellogenesis in the Kuruma Prawn, Penaeus japonicus.

In Crustacea, reproductive function and mechanisms regulating vitellogenesis have not been fully elucidated. This is due in great part to a lack of information concerning the biochemical nature of the vitellogenin molecule, the hemolymph precursor of yolk protein, vitellin, as well as the functional expression of the vitellogenin-encoding gene. We have therefore cloned a cDNA encoding vitellogenin in the kuruma prawn, Penaeus japonicus based on the N-terminal amino acid sequence of the 91 kDa subunit of vitellin. The open reading frame of this cDNA encoded 2,587 amino acid residues. This is the first investigation reporting a full-length cDNA and its corresponding amino acid sequence for vitellogenin in any crustacean species.Northern blot analysis and in situ hybridization have revealed that mRNA encoding vitellogenin was expressed in both the follicle cells in the ovary and the parenchymal cells in the hepatopancreas. In nonvitellogenic females, vitellogenin mRNA levels were negligible in both the ovary and hepatopancreas, but in vitellogenic females, levels were dramatically increased in both tissues. In the ovary, highest levels were observed during the early exogenous vitellogenic stage, and thereafter rapidly decreased, whereas in the hepatopancreas, high levels were maintained until the onset of the late vitellogenic stage. Differing profiles of vitellogenin mRNA levels in the ovary and hepatopancreas suggest that the contribution of these tissues to vitellogenin synthesis harbor separate and complementary roles during vitellogenesis.

Mouse popliteal lymph node assay for assessment of allergic and autoimmunity-inducing potentials of low-molecular-weight drugs.

In the present collaborative study, popliteal lymph node (PLN) responses to penicillin G (an allergenic chemical), D-penicillamine (an autoimmunity-inducing chemical), and barbital (a negative reference chemical) were investigated in three different mouse strains by ten pharmaceutical companies. Two inbred mouse strains (BALB/c and A/J) and one outbred strain (ICR) were subcutaneously injected with saline solutions containing penicillin G (1.25, 2.5 and 5 mg/mouse), D-penicillamine (0.5, 1 and 2 mg/mouse), or barbital (2 mg/mouse) into one hind footpad and saline only was injected into the contralateral footpad. PLN cellularity indices were determined on day 7. In the three strains tested, the penicillin G and D-penicillamine injections resulted in approximately dose-dependent responses. In contrast, barbital failed to generate a significant PLN reaction. In the typical data from one of the participating laboratories, the PLN responses of A/J, BALB/c, and ICR to penicillin G were high, intermediate and low, respectively, while their PLN responses to D-penicillamine were all high. Some variation in PLN cellularity indices was observed among the participating laboratories, but reproducibility of the popliteal lymph node assay (PLNA) evaluation was partly confirmed. Although the appropriate selection of mouse strains and drug dosage levels has to be considered, these results suggest that the PLNA may be an appropriate screening system for prediction of the allergic or autoimmunity-inducing potentials of low-molecular-weight drugs.

Transforming growth factor beta-induced failure of resistance to infection with blood-stage Plasmodium chabaudi in mice.

The role of transforming growth factor beta (TGF-beta) in infection with Plasmodium chabaudi was investigated with resistant and susceptible mouse models. C57BL/10 mice produced gamma interferon (IFN-gamma) and nitric oxide (NO) shortly after infection and cleared the parasite spontaneously. In contrast, BALB/c mice showed a transient enhancement of TGF-beta production, followed by a relative lack of IFN-gamma and NO production, and succumbed to the infection. However, there was no correlation between levels of serum TGF-beta and splenic TGF-beta mRNA in both mouse strains before and after infection. Administration of recombinant TGF-beta (rTGF-beta) rendered resistant mice susceptible because of suppression of subsequent production of IFN-gamma and NO. Administration of anti-TGF-beta antibody to the infected BALB/c mice resulted in remarkable increases in serum IFN-gamma and NO, and the mice resisted the infection. Splenic CD4(+) T and CD11b+ cells of C57BL/10 mice were significantly activated after infection, but this was completely abrogated by administration of rTGF-beta. These results suggested that, in the P. chabaudi-susceptible but not resistant mice, production of TGF-beta was promoted, and subsequent failure of IFN-gamma- and NO-dependent resistance to the parasite was induced. This study is the first to indicate that TGF-beta production was the key event in failure of resistance to mouse malaria.

Suppression of in vitro IFN-gamma production by spleen cells of Plasmodium chabaudi-infected C57BL/10 mice exposed to dexamethasone at a low dose.

After infection with Plasmodium chabaudi, C57BL/10 mice produced significant amounts of serum IFN-gamma, developed a low level of parasitemia and survived the infection. Production of IFN-gamma was also obvious when spleen cells of the infected mice were stimulated with the parasite antigen contained in an erythrocyte lysate in vitro. Depletion of CD4+ T cells abrogated production of IFN-gamma, leading to loss of resistance to the infection. The CD4+ T cells/IFN-gamma-dependent resistance of the C57BL/10 mice against P. chabaudi was applied to assess immunotoxicological effect of dexamethasone (DEX). Administration of a high dose (0.75 mg/kg) resulted in loss of the splenic cellularity, remarkable decrease in serum IFN-gamma production, increased level of parasitemia, and eventual death of the infected mice. In contrast, DEX at a low dose (0.02 mg/kg) induced no alternation in the in vivo host immune activity and the mice survived the infection. However, when spleen cells were obtained from the infected mice administered the low dose of DEX and stimulated in vitro with the parasite antigen, a significantly decreased level of IFN-gamma was demonstrated with compared to control mice. These findings demonstrate that the in vitro production of IFN-gamma by spleen cells from P. chabaudi-resistant C57BL/10 mice was more sensitive to the immunosuppressive effect of in vivo administration of DEX. This ex vivo assay might provide a method for evaluation of drug-induced immunotoxicity at a higher sensitivity than the conventional host resistant assays such as comparison of severity of disease or time to death.

Protein-nutritive assessment of sake lees obtained by brewing from liquefied rice.

Sake lees obtained by brewing from liquefied rice were deprived of water and alcohol by lyophilization, and then examined for nutritional availability with the aid of proximate food analysis, amino acid analysis and animal experiment. Freeze-dried sake lees powder was comprised of 44.6% protein, 37.4% carbohydrate, 2.5% fat, 6.7% fiber, 1.8% ash and 7.2% moisture (alcohol < 0.1%), of which the nutritive value (amino acid score) was estimated as 89.6 when compared with the amino acid requirements for preschool children (FAO/WHO/UNU, 1985). Sake lees protein had been, however, appreciably improved in the limiting amino acid "lysine" relative to polished rice protein. As a result of an animal experiment, the rats fed a 50% sake lees powder diet proved to be equal in growth to those fed a 20% casein (control) diet, although the former diet had to be supplemented with vitamins and minerals, which were in shortage as compared to the control diet. On the other hand, the feeding of sake lees powder was effective in lowering the serum triacylglycerol concentration. Accordingly, sake lees powder can be assessed as a favorable candidate for not only protein-rich but also hypolipidemic provisions.

Solubilization and chemical characterization of an insoluble matrix protein in the gastroliths of a crayfish, Procambarus clarkii.

The gastrolith of the crayfish Procambarus clarkii contains a small amount of an organic matrix that is mainly chitin and proteins, together with a large amount of calcium carbonate. As the first step to understand the mechanism of calcification, we tried to characterize matrix proteins in the gastrolith. An insoluble matrix protein, referred to as gastrolith matrix protein, was made soluble with 1% SDS containing 10 mM dithiothreitol, and was purified by reverse-phase high-performance liquid chromatography. The protein had a molecular weight of about 50,500 and a blocked amino terminus. By enzymatic digestion and microsequencing, five partial amino acid sequences with a total of 225 amino acid residues were identified and found to include a repetitive sequence not reported previously.