Potential for populations of Aedes j. japonicus to transmit Rift Valley fever virus in the USA.

Aedes japonicus japonicus was introduced into the northeastern USA in 1998 and has since spread to more than 25 states. Because this species has been shown to be a competent laboratory vector of several viruses, readily feeds on large mammals, and has become a pest in several areas, there is concern that it might serve as a vector of Rift Valley fever virus (RVFV) should that virus be introduced into North America. Infection with RVFV causes mortality in > 90% of young domestic ungulates (e.g., calves, kids, and lambs), as well as causing a febrile illness and occasional deaths in humans. Therefore, we evaluated Ae. j. japonicus captured in North Carolina and in Maryland for their ability to serve as potential vectors for RVFV. After feeding on infected adult hamsters, these mosquitoes were tested for infection, dissemination, and the ability to transmit RVFV after incubation at 26 degrees C for 7-28 days. Both the Maryland and North Carolina populations of Ae. j. japonicus were highly efficient laboratory vectors of RVFV, with infection rates > 90% and dissemination rates > 84% for those mosquitoes that fed on hamsters with viremias > or = 10(8.5) plaque-forming units/ml. Thus, Ae. j. japonicus should be targeted for immediate control should RVFV be introduced into an area where this mosquito is now present.

LX4211, a dual SGLT1/SGLT2 inhibitor, improved glycemic control in patients with type 2 diabetes in a randomized, placebo-controlled trial.

Thirty-six patients with type 2 diabetes mellitus (T2DM) were randomized 1:1:1 to receive a once-daily oral dose of placebo or 150 or 300 mg of the dual SGLT1/SGLT2 inhibitor LX4211 for 28 days. Relative to placebo, LX4211 enhanced urinary glucose excretion by inhibiting SGLT2-mediated renal glucose reabsorption; markedly and significantly improved multiple measures of glycemic control, including fasting plasma glucose, oral glucose tolerance, and HbA(1c); and significantly lowered serum triglycerides. LX4211 also mediated trends for lower weight, lower blood pressure, and higher glucagon-like peptide-1 levels. In a follow-up single-dose study in 12 patients with T2DM, LX4211 (300 mg) significantly increased glucagon-like peptide-1 and peptide YY levels relative to pretreatment values, probably by delaying SGLT1-mediated intestinal glucose absorption. In both studies, LX4211 was well tolerated without evidence of increased gastrointestinal side effects. These data support further study of LX4211-mediated dual SGLT1/SGLT2 inhibition as a novel mechanism of action in the treatment of T2DM.

Updated distribution records for Anopheles vagus (Diptera: Culicidae) in the Republic of Philippines, and considerations regarding its secondary vector roles in Southeast Asia.

Distribution records for Anopheles (Cellia) vagus in the Republic of the Philippines were updated, including recent collection and museum records from Luzon and Visayas Provinces. Larval habitats (e.g. rice paddies, irrigation and drainage ditches), associated species, and the vector potential of this species were also noted.

Serum vasopressin concentrations during orthotopic liver transplantation.

BACKGROUND: We report measurements of the temporal response of serum vasopressin concentrations in the period after reperfusion of the liver graft during orthotopic liver transplantation (OLT). METHODS: Vasopressin concentrations were determined in 11 adult patients undergoing OLT by radioimmunoassay of samples collected after induction, at 5 minutes prior to reperfusion, and at 10, 20, 30, 40, 50, 60, 90, and 120 minutes after reperfusion. RESULTS: Pre-incision vasopressin concentrations ranged from <0.5 to 2.6 pg/mL (reference serum vasopressin, <1.7 pg/mL). Overall, levels increased before reperfusion, but fell thereafter. Individual patients manifested elevated levels during the period after reperfusion. Values immediately before reperfusion exhibited most variability, ranging from 0.8 to 40 pg/mL (median, 15; interquartile range [IQR], 4-29) Median vasopressin concentrations 10 minutes postreperfusion were 7.6 pg/mL (IQR, 3-27). Only 3 of the 11 patients failed to generate vasopressin levels >20 pg/mL. In each of these patients, hemodynamics were satisfactory without the need for additional pressor infusion. Maximum vasopressin concentration measured in any patient was 85 pg/mL. There was no correlation between vasopressin concentration and mean blood pressure or systemic vascular resistance index. CONCLUSION: Vasopressin concentrations during OLT vary widely and are elevated periodically during the anhepatic and postreperfusion stages, with no apparent relationship between vasopressin concentrations and blood pressure. Although vasopressin concentrations were not as high as those measured during some other clinical situations, these data suggest that a relative vasopressin deficiency is not a direct cause of hypotension during OLT.

Influence of coronary artery disease on outcomes after liver transplantation.

Patients with coronary artery disease (CAD) who undergo liver transplantation (OLT) have been previously identified as a high-risk group. Since that identification, the management of CAD has undergone significant changes as has the cardiovascular screening and selection of patients for OLT. We retrospectively identified 42 patients with known CAD who underwent OLT to compare outcomes with a control group of 42 patients without CAD who were matched for gender, age, and primary liver disease. Mortality rates were higher in the CAD than the control group at 1 year (5 vs 1) and 3 years (11 vs 3; P < .05) although lower than previously reported (at 3 years, 26% vs 50%). New cardiovascular morbidity was also more frequent among the CAD than control group at 1 year (11 vs 3; P < .05) and 3 years (16 vs 4; P < .05). Although outcomes for patients with CAD undergoing OLT are improved from historical levels, they are still worse than those in patients without CAD despite current management and selection strategies.

Effect of lidocaine on endothelin-1-mediated airway smooth muscle contraction in the rat trachea.

BACKGROUND: Endothelin-1 (ET-1) is a 21-amino acid peptide that induces airway smooth muscle (ASM) constriction by activating G-protein-coupled endothelin receptors A (ETA) and B (ETB), thereby increasing intracellular calcium ([Ca(2+)]i). Lidocaine can cause direct ASM relaxation by decreasing [Ca(2+)]i. This study investigated the direct relaxant effects of lidocaine on ET-1-induced contraction in rat tracheas. METHODS: Mid-tracheal rings (2-3 mm diameter) were excised and attached to a force transducer suspended in Krebs-Henseliet solution. Carbachol concentration-response curves (10 nM and 100 microM) were generated to determine maximal contractility (C(max)). ET-1 (3 nM to 200 nM) responses to lidocaine (100 nM, 10 microM, and 1 mM) were measured in the presence and absence of extracellular calcium. Contractile responses to ET-1 are presented as percentage of Cmax (% Cmax). Data were analyzed using analysis of variance and unpaired t-tests with Welsh correction. RESULTS: No significant effect on ET-1-induced constriction was noted in the presence of low concentrations of lidocaine (100 nM and 1 muM), with and without extracellular calcium. At a concentration of 1 mM, lidocaine decreased the response to 100 nM and 200 nM ET-1 by 26% in the presence of extracellular calcium and by 37 and 44%, respectively, in the absence of calcium. CONCLUSION: The attenuating effect of lidocaine (1 mM) on ET-1-induced ASM contraction is not exclusively dependent on the blockade of intracellular calcium entry.

Occurrence and relative abundance of mosquitoes in stormwater retention facilities in North Carolina, U.S.A.

Throughout the 2004 mosquito season, 52 stormwater retention facilities were sampled to characterize the seasonal occurrence and relative abundance of mosquito species in relation to the structural complexity and biological diversity of the facilities. The three different types of facilities included standard wet ponds (n=20), innovative ponds (n=14), and wetland ponds (n=18). All retention structures were sampled at the beginning, middle and end of the mosquito season so that seasonal changes in mosquito production could be characterized. Overall samplings, mosquitoes were collected from 34% of the retention structures. Fourteen species representing 7 genera were collected, but only 5 species were commonly collected in all three types of stormwater management facilities. In general, the seasonal prevalence and relative abundance of mosquito species did not vary among three types of retention structures. A significant association (P < 0.01) between the presence of mosquito larvae or pupae and the absence of mosquitofish was found for innovative and wetland stormwater retention facilities but not for standard retention facilities (P > 0.05).

Identification of selective inhibitors of cyclin dependent kinase 4.

A new structural type of kinase inhibitor, containing a benzocarbazole nucleus, has been identified. Members of the series are selective for inhibition of the cyclin dependent kinase family of enzymes. Although the cdks are highly homologous, representatives of the series showed intra-cdk selectivities, especially for cdk4. SAR studies elucidated the important features of the molecules for inhibition.

The synthesis of an exhaustively stereodiversified library of cis-1,5 enediols by silyl-tethered ring-closing metathesis.

[reaction: see text] This report describes the parallel synthesis of all 16 stereoisomers of the cis-1,5 enediol module 1. Compounds 1 derive from 2 by silicon-tethered ring-closing metathesis. Such libraries of stereodiversified ligands provide a unique approach to ligand discovery that employs exhaustive searching of conformational space.

Evaluation of 1-octen-3-ol, carbon dioxide, and light as attractants for mosquitoes associated with two distinct habitats in North Carolina.

Field studies were conducted in North Carolina to determine the responses of mosquitoes found in salt marsh and inland creek flood plain areas to 1-octen-3-ol (octenol), carbon dioxide (CO2), and light in various combinations with Centers for Disease Control (CDC) light traps. Over 56,000 adult mosquito specimens of 12 species in 4 genera were collected in the salt marsh. They exhibited a general response pattern of octenol + CO2 + light > CO2 + light = octenol + CO2 > octenol + light > octenol alone. Significantly, more Aedes sollicitans, Ae. taeniorhynchus, Anopheles bradleyi, and Culex salinarius were attracted to octenol + CO2 + light than to CO2 + light. Over 19,000 specimens of 24 species in 7 genera were collected in the inland creek flood plain. Although the response patterns to the attractants were similar to those in the salt marsh area, there was no significant difference between octenol + CO2 + light and CO2 + light. Aedes vexans, An. crucians, and An. punctipennis were attracted nearly equally to these two attractant combinations. These studies demonstrate that responses to combinations of these attractants are species specific. However, different combinations of attractants can significantly increase the collection of targeted species important in arbovirus transmission. The use of these combinations would be very beneficial in mosquito-borne virus surveillance studies. The use of octenol by itself or in conjunction with light was found the least useful for collecting mosquitoes in both habitats.

Galectin-3-mediated adherence of Proteus mirabilis to Madin-Darby canine kidney cells.

Proteus mirabilis is an important cause of urinary tract infections (UTIs) and can result in acute pyelonephritis. Proteus mirabilis expresses several, morphologically distinct, fimbrial species, and previous studies have shown that the nonagglutinating fimbriae (NAF) mediate bacterial adherence to a number of cell lines, including Madin-Darby canine kidney (MDCK) cells. Immunoblot overlay analysis of the plasma membrane fraction from MDCK cells with purified NAF revealed a 34-kDa band, which has been analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Database search identified galectin-3 as a potential protein candidate. Immunocytochemical assay of MDCK cells with a galectin-3-specific monoclonal antibody, anti-Mac-2, confirmed its presence on the plasma membrane extracellular surface. Preincubation of P. mirabilis with anti-Mac-2 monoclonal antibodies, specific for galectin-3, resulted in the inhibition of bacterial binding to MDCK cells. These data suggest a role for galectin-3, interacting with appropriately glycosylated surface receptors and P. mirabilis fimbriae, as a mediator of bacterial adherence in vitro.

The binding of Proteus mirabilis nonagglutinating fimbriae to ganglio-series asialoglycolipids and lactosyl ceramide.

Proteus mirabilis is a common opportunistic Gram-negative uropathogen that infects the upper urinary tract. We have examined the role of the nonagglutinating fimbriae (NAF) of P. mirabilis in mediating bacterial adhesion to cell surface receptors. Purified NAF of P. mirabilis were demonstrated to bind to a number of glycolipids, including asialo-GM1, asialo-GM2, and lactosyl ceramide (LacCer) in solid-phase binding assays and in thin layer chromatography (TLC) overlay assays. Furthermore, preincubation of the biotinylated NAF (Bt-NAF) with anti-NAF monoclonal antibodies resulted in inhibition of NAF binding to immobilized asialo-GM1, asialo-GM2, and LacCer. In adherence assays, P. mirabilis binding to Madin-Darby canine kidney (MDCK) cells was inhibited by murine anti-asialo-GM1 monoclonal antibodies H2G10 to about 50% of the binding level in the absence of the antibody, specific for the terminal beta-galactopyranosyl residue of asialo-GM1 (Harrison et al. 1998). The results of this study suggest that NAF of P. mirabilis recognize a GalNAc beta 1-4Gal moiety present in the ganglio-series of asialoglycolipids, and that the terminal beta-galactopyranosyl-containing glycoconjugates play a role in NAF-mediated adherence of P. mirabilis to MDCK cells. Similarly to other bacteria, P. mirabilis NAF was also shown to express the LacCer specificity.

Inhaled nitric oxide reduces pulmonary artery pressures in portopulmonary hypertension.

Pulmonary artery hypertension in association with liver failure (portopulmonary hypertension [PPHTN]) is a significant barrier to liver transplantation because patients with this condition have a very high mortality when transplantation is undertaken. Inhaled nitric oxide (NO), a potent pulmonary vasodilator, reduces pulmonary artery pressure (PAP) in some patients with primary pulmonary hypertension, but its effect in patients with PPHTN is controversial. We investigated the hemodynamic effects of inhaled NO in 6 patients with PPHTN. Five of 6 patients responded to NO inhalation with decreases in PAP and pulmonary vascular resistance of greater than 10%; these decreases were statistically significant at NO concentrations of 10 and 30 ppm. Cardiac output did not significantly change. We conclude that inhalation of NO reduces PAPs in some patients with PPHTN.

Preclinical antitumor efficacy of analogs of XK469: sodium-(2-[4-(7-chloro-2-quinoxalinyloxy)phenoxy]propionate.

A series of quinoxaline analogs of the herbicide Assure was found to have selective cytotoxicity for solid tumors of mice in a disk-diffusion-soft-agar-colony-formation-assay compared to L1210 leukemia. Four agents without selective cytotoxicity and 14 agents with selective cytotoxicity were evaluated in vivo for activity against a solid tumor. The four agents without selective cytotoxicity in the disk-assay were inactive in vivo (T/C > 42%). Thirteen of the fourteen agents with selectivity in the disk-assay were active in vivo (T/C < 42%). Five of the agents had curative activity. These five agents had a halogen (F, Cl, Br) in the 7-position (whereas Assure had a CI in the 6 position). All agents with curative activity were either a carboxylic acid, or a derivative thereof, whereas Assure is the ethyl ester of the carboxylic acid. All other structural features were identical between Assure and the curative agents. Assure had no selective cytotoxicity for solid tumors in the disk-assay, and was devoid of antitumor activity. The analog XK469 is in clinical development.

A kinetics approach to the characterization of an IgM specific for the glycolipid asialo-GM1.

The unique features of protein recognition of membrane-anchored glycolipids were investigated by surface plasmon resonance (SPR) monitoring of antibody interactions with glycolipids contained in liposomes. Several positive hybridomas belonging to the IgM and IgG classes were identified when tested for binding to the glycosphingolipid asialo-GM1 (Gal beta1-3GalNAcl beta1-4Gal beta1-4Glc beta1-1-Ceramide). Preliminary screening by enzyme immunoassay and thin layer chromatography (TLC) followed by immunostaining indicated that only those of the IgM type showed specificity for this glycosphingolipid. One of the IgMs, H2G10, was purified and further characterized using a SPR technique that involved antibody binding to liposomal asialo-GM1. This method generated kinetic and affinity constants for the interaction and confirmed the specificity of H2G10 for the terminal galactose of asialo-GM1. Interestingly, inhibition of antibody binding to asialo-GM1 liposomes by the asialo-GM1 tetrasaccharide reduced the total amount of bound antibody but increased the affinity of the antigen-antibody interaction due to an inverse relationship between tetrasaccharide concentration and the H2G10 dissociation rate constant. We believe that this effect is due to the selective inhibition of lower valency binding by the tetrasaccharide which, in turn, promotes higher avidity antibody-carbohydrate interactions. The observation that slower dissociation rate constants were also observed at high antigen to antibody ratios supports this interpretation. These results highlight the insight that kinetic data can provide in efforts to promote and inhibit high avidity interactions such as those involving proteins and carbohydrates.

North Carolina mosquito records. I. Uncommon Aedes and Anopheles (Diptera: Culicidae).

New distribution records are provided for 10 mosquito species that are rare or uncommon in North Carolina: Aedes aegypti, Ae. cinereus, Ae. dupreei, Ae. fulvus pallens, Ae. hendersoni, Ae. mitchellae, Ae. thibaulti, Ae. tormentor, Ae. trivittatus, and Anopheles atropos. Biological notes are provided for habitats, behavior, and, in some cases, color patterns. Comments are also made about 6 additional species that are rare or uncommon in North Carolina.

The expression of nonagglutinating fimbriae and its role in Proteus mirabilis adherence to epithelial cells.

Proteus mirabilis is a common causative agent of human urinary tract infections, especially in catheterized patients and in those patients with structural abnormalities of the urinary tract. In addition to the production of hemolysin and urease, fimbriae-mediated adherence to uroepithelial cells and kidney epithelium may be essential for virulence of P. mirabilis. A single P. mirabilis strain is capable of expressing several morphologically distinct fimbrial species, which can each be favoured by specific in vitro growth conditions. The fimbrial species reported to date include mannose-resistant/Proteus-like fimbriae, ambient temperature fimbriae, P. mirabilis fimbriae, and nonagglutinating fimbriae (NAF). Here, using intact bacteria or purified NAF as immunogens, we have generated the first reported NAF-specific monoclonal antibodies (mAbs). Bacteria expressing NAF as their only fimbrial species adhered strongly to a number of cell lines in vitro, including uroepithelial cell lines. Binding of P. mirabilis was markedly reduced following preincubation with NAF-specific mAbs and Fab fragments. The presence of NAF with highly conserved N-terminal sequences on all P. mirabilis strains so far examined, combined with the ability of both anti-NAF mAbs and purified NAF molecules to inhibit P. mirabilis adherence in vitro, suggests that NAF may contribute to the pathogenesis of P. mirabilis.

Ticks (Acari: Ixodida) uncommonly found biting humans in North Carolina.

Collection records are presented that document human-biting by Otobius megnini, Amblyomma maculatum, Haemaphysalis leporispalustris, Ixodes cookei, Ixodes dentatus, and Rhipicephalus sanguineus in North Carolina. These species are either extremely rare in North Carolina or they are normally considered non-human feeders. The record of Otobius megnini represents the first collection of this species in North Carolina in over 50 years. It is proposed that immature Rhipicephalus sanguineus feed on humans much more than previously suspected and that they represent a threat for the transmission of pathogens to humans.

Identifying Psorophora horrida females in North Carolina (Diptera:Culicidae).

Morphological characters to separate Psorophora horrida (Dyar and Knab) and Psorophora ferox (Von Humboldt) females are identified and defined. In addition to the 3 characters used to separate these 2 species in published keys, an additional 6 characters were found. Of the total of 9 characters identified, 6 are reliable at the 100% level, based on North Carolina specimens. These additional characters will assist greatly in identifying Ps, horrida specimens. Variation or hindtarsomere 4 of Ps. horrida, consisting of dark-scaled patterns, were also noted. Characters for separating Psorophora cyanescens (Coq.) and Psorophora mathesoni Belkin and Heinemann from Ps. ferox and Ps. horrida are discussed.