Ivermectin performance in horses diagnosed with equine endocrine disorders.

Anthelmintic performance against equine cyathostomins can be evaluated by two different non-terminal measures; the Fecal Egg Count Reduction Test (FECRT) and the Egg Reappearance Period (ERP). Most available FECRT and ERP data have been determined in populations of young horses, and very little information is available from mature and senior horses. Furthermore, it is unknown how commonly occurring equine endocrine disorders such as Insulin dysregulation (ID) and Pituitary pars intermedia dysfunction (PPID) may interfere with these measurements, but it has been suggested that horses with these conditions could be more susceptible to parasitic infections. A research population of senior horses and horses with or without PPID, ID, or both were enrolled in this study. All strongylid egg count positive horses were included in an ivermectin (200 µg/kg) efficacy study. These were distributed among the following groups: ID: six, PPID: three, PPID and ID: seven, and healthy controls: three. Strongylid fecal egg counts were determined on the day of ivermectin administration, at two weeks post deworming, and on weekly intervals until eight weeks post treatment. Determination of FECRT and ERP were carried out following World Association for the Advancement of Veterinary Parasitology guidelines. Results revealed high ivermectin efficacy with mean egg count reduction at 99.7% or above in all groups at two weeks post treatment. Egg reappearance was documented at six and seven weeks in the ID and PPID/ID groups, respectively, whereas the PPID and healthy control groups both had ERP at 8 weeks. Statistical analysis found no significant differences in egg count levels between groups during the study. The expected ERP for ivermectin is 8-10 weeks, meaning that two of the groups displayed shortened ERPs. However, due to the small group sizes, these data should be interpreted with caution. Nonetheless, results do indicate a need for further investigation of the possible influence of endocrine disorders on anthelmintic performance in horses.

Wellbeing and mental health outcomes amongst hospital healthcare workers during COVID-19.

BACKGROUND: Global healthcare systems have been particularly impacted by the COVID-19 pandemic. Healthcare workers (HCWs) are widely reported to have experienced increased levels of baseline psychological distress relative to the general population, and the COVID-19 pandemic may have had an additive effect. However, previous studies are typically restricted to physicians and nurses with limited data available on hospital HCWs. We aimed to conduct a cross-sectional, psychological evaluation of Irish HCWs during COVID-19. METHODS: HCWs across five adult acute level-4 Dublin-based hospitals completed an online survey of wellbeing and COVID-19 experience. RESULTS: There were 1898 HCWs who commenced the survey representing 10% of the total employee base. The sample comprised nurses (33%), doctors (21%), Health and Social Care Professionals (HSCPs) (24%) and 'Other' disciplines (22%), and 81% identified as female. Clinical levels of depression, anxiety and PTSD symptoms were endorsed by 31%, 34% and 28% of respondents, respectively. Professional grouping effects included: nurses reporting significantly greater levels of COVID-19 exposure, infection, COVID-fear, moral injury, and post-traumatic distress; HSCPs were significantly less likely to report mood dysfunction. In terms of gender, males were significantly less likely to report negative pandemic experiences, low resilience, and significantly more likely to endorse 'minimal' depression, anxiety, and traumatic distress. Logistic regression modelling revealed mental health outcomes (depression, anxiety and PTSD symptoms) were associated with increased frontline exposure, fewer career years' experience, elevated pre-pandemic stress, and female gender. DISCUSSION: To our knowledge, this is the largest evaluation of psychological wellbeing amongst HCWs in acute hospitals in the Dublin region. Our findings have implications for healthcare workforce wellbeing and future service delivery.

The effect of obesity on adverse outcomes and metabolism in pediatric burn patients.

HYPOTHESIS: Obesity influences metabolism and increases the incidence of clinical complications and worsens outcomes in pediatric burn patients. DESIGN: Retrospective, single-center study. SUBJECTS: In all, 592 severely burned pediatric patients who had burns covering more than 30% of the total body surface area and who were treated between 2001 and 2008 were enrolled in this study. Patients were divided into ≥85th percentile (n=277) and normal (n=315) weight groups based on body mass index (BMI) percentiles. RESULTS: Patients stratified below (normal) and ≥85th percentile had similar age, gender distribution and total burn size. No significant differences were detected in the incidence of sepsis (11% for obese vs 10% for normal), the incidence of multiple organ failure (MOF) (21% for obese and 16% for normal) or mortality (11% for obese vs 8% for normal). Compared with the normal group, the ≥85th percentile group had low levels of constitutive proteins (α2macroglobulin and Apolipoprotein A1) (P<0.05 for both) as well as high levels of triglycerides and the acute-phase protein, C-reactive protein (P<0.05 for both) up to 60 days after injury. Patients ≥85th percentile showed a significant higher loss of bone mineral density and lipolysis compared with normal individuals. Stepwise logistic regression analysis revealed that BMI had a positive predictive value towards the maximum DENVER2 score, an index of organ failure (P<0.001). CONCLUSIONS: BMI≥85th percentile altered the post-burn acute phase and catabolic response but did not increase the incidence of sepsis, MOF or mortality in pediatric burn patients. Our results suggest that impaired metabolism and an altered inflammatory response already exists in patients starting at the 85th percentile BMI.

The structure and composition of liposomes can affect skin regeneration, morphology and growth factor expression in acute wounds.

Liposomal gene transfer is an effective therapeutic approach to improve dermal and epidermal regeneration. The purpose of the present study was to define whether the biological or chemical structure of a liposome influences cellular and biological regeneration in the skin, and to determine by which mechanisms possible changes occur. Rats were inflicted a full-excision acute wound and divided into three groups to receive weekly subcutaneous injections of DMRIE liposomes plus the Lac Z gene, or DOTAP/Chol liposomes plus the Lac Z gene, or saline. Planimetry, immunological assays, histological and immunohistochemical techniques were used to determine cellular responses after gene transfer, protein expression, dermal and epidermal regeneration. DOTAP/Chol increased IGF-I and KGF protein concentration and caused concomitant cellular responses, for example, by increasing IGFBP-3, P<0.05. DOTAP/Chol liposomes improved epidermal regeneration by exhibiting the most rapid area and linear wound re-epithelization compared to DMRIE or control, P<0.001. DOTAP/Chol and DMRIE exerted promitogenic and antiapoptotic effects on basal keratinocytes, P<0.05. Dermal regeneration was improved in DOTAP/Chol-treated animals by an increased collagen deposition and morphology, P<0.001. DOTAP/Chol liposomes increased vascular endothelial growth factor concentrations and thus neovascularization when compared with DMRIE and saline, P<0.001. In the present study, we showed that different liposomes have different effects on intracellular and biological responses based on its chemical and molecular structure. For gene transfer in acute wounds, the administration of DOTAP/Chol liposomes appears to be beneficial.

Biochemical and crystallographic characterization of ferredoxin-NADP(+) reductase from nonphotosynthetic tissues.

Distinct forms of ferredoxin-NADP(+) reductase are expressed in photosynthetic and nonphotosynthetic plant tissues. Both enzymes catalyze electron transfer between NADP(H) and ferredoxin; whereas in leaves the enzyme transfers reducing equivalents from photoreduced ferredoxin to NADP(+) in photosynthesis, in roots it has the opposite physiological role, reducing ferredoxin at the expense of NADPH mainly for use in nitrate assimilation. Here, structural and kinetic properties of a nonphotosynthetic isoform were analyzed to define characteristics that may be related to tissue-specific function. Compared with spinach leaf ferredoxin-NADP(+) reductase, the recombinant corn root isoform showed a slightly altered absorption spectrum, a higher pI, a >30-fold higher affinity for NADP(+), greater susceptibility to limited proteolysis, and an approximately 20 mV more positive redox potential. The 1.7 A resolution crystal structure is very similar to the structures of ferredoxin-NADP(+) reductases from photosynthetic tissues. Four distinct structural features of this root ferredoxin-NADP(+) reductases are an alternate conformation of the bound FAD molecule, an alternate path for the amino-terminal extension, a disulfide bond in the FAD-binding domain, and changes in the surface that binds ferredoxin.

Characterization of a granulovirus from the cassava hornworm (Erinnyis ello: Sphingidae).

A Colombian isolate of Erinnyis ello granulovirus (EeGV) was characterized by electron microscopy, restriction endonuclease digestion, and SDS-PAGE. Electron microscopy showed the occlusion bodies to have a morphology typical of granuloviruses. The restriction patterns of DNA from EeGV and the granuloviruses of Trichoplusia ni (TnGV) and Pieris rapae (PrGV) show little or no similarity, indicating little relatedness among these viruses. EeGV was estimated to possess a relatively small genome of 90.5 +/- 0.5 kbp. SDS-PAGE analysis compared the occulsion body and enveloped nucleocapsid proteins of EeGV and TnGV, and the polypeptide patterns also showed little similarity between these viruses. These analyses, as well as comparison of our results to those reported for other granuloviruses, indicate that EeGV represents a new granulovirus isolate.

Host cell receptor binding by baculovirus GP64 and kinetics of virion entry.

GP64 is the major envelope glycoprotein from budded virions of the baculoviruses Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) and Orgyia pseudotsugata multicapsid nucleopolyhedrovirus (OpMNPV). To examine the potential role of GP64 as a viral attachment protein in host cell receptor binding, we generated, overexpressed, and characterized a soluble form of the OpMNPV GP64 protein, GP64solOp. Assays for trimerization, sensitivity to proteinase K, and reduction by dithiothreitol suggested that GP64solOp was indistinguishable from the ectodomain of the wild-type OpMNPV GP64 protein. Virion binding to host cells was analyzed by incubating virions with cells at 4 degrees C in the presence or absence of competitors, using a single-cell infectivity assay to measure virion binding. Purified soluble GP64 (GP64solOp) competed with a recombinant AcMNPV marker virus for binding to host cells, similar to control competition with psoralen-inactivated wild-type AcMNPV and OpMNPV virions. A nonspecific competitor protein did not similarly inhibit virion binding. Thus specific competition by GP64solOp for virion binding suggests that the GP64 protein is a host cell receptor-binding protein. We also examined the kinetics of virion internalization into endosomes and virion release from endosomes by acid-triggered membrane fusion. Using a protease sensitivity assay to measure internalization of bound virions, we found that virions entered Spodoptera frugiperda Sf9 cells between 10 and 20 min after binding, with a half-time of approximately 12.5 min. We used the lysosomotropic reagent ammonium chloride to examine the kinetics of membrane fusion and nucleocapsid release from endosomes after membrane fusion. Ammonium chloride inhibition assays indicated that AcMNPV nucleocapsids were released from endosomes between 15 and 30 min after binding, with a half-time of approximately 25 min.

The insect immune protein scolexin is a novel serine proteinase homolog.

Scolexin is a coagulation-provoking plasma protein induced in response to bacterial or viral infection of larval Manduca sexta, a large lepidopterous insect. Here we report the isolation and sequencing of two cDNA clones that code for scolexin isoforms sharing 80% sequence identity. The scolexin sequences have low but recognizable sequence similarity to members of the chymotrypsin family and represent a new subfamily of chymotrypsin-like serine proteinases. Comparison with known structures reveals the conservation of key catalytic residues and a possible specificity for small nonpolar residues. Most remarkable is the absence of a canonical activation peptide cleavage site. This suggests that the regulation of scolexin activity will involve a novel activation mechanism.

The plasma protein scolexin from Manduca sexta is induced by baculovirus infection and other immune challenges.

Manduca sexta larvae infected per os with a sublethal dose of Erinnyis ello granulosis virus (EeGV) contain an induced plasma protein of ca 33-36 kDa [Finnerty et al. (1994) J. Invertebr. Pathol. 63, 140-144]. This virus-induced protein shares characteristics with scolexin, a bacteria-induced plasma protein from M. sexta, so in this study we compared the two proteins. Two-dimensional gel electrophoresis reveals the induction of polypeptides corresponding to the subunits of scolexin in the plasma from EeGV-infected, as well as bacteria-injected, larvae. Immunoblots of these two-dimensional polyacrylamide gels show that antiserum against the virus-induced protein cross-reacts with the putative scolexin polypeptides induced by either EeGV infection or bacteria injection. Amino acid analysis of the virus-induced protein shows it to be very similar to scolexin, and the N-terminal sequences of the two proteins are nearly identical. From these data, we conclude that the virus-induced protein and scolexin are identical, or at least isoforms of the same protein. Using immunoblot, we also demonstrate that scolexin is induced in M. sexta plasma by injection with yeast or lipopolysaccharide from Serratia marcescens.

Effects of chronic administration of benfluorex to rats on the metabolism of corticosterone, glucose, triacylglycerols, glycerol and fatty acid.

(1) Rats were fed on diets enriched with sucrose, beef tallow or corn oil and treated for 11-16 days with 50 mg of benfluorex per kg of body weight. By these times the growth rate and food intake were not significantly different from those of control rats. (2) Benfluorex approximately halved the concentration of circulating triacylglycerol in rats fed the beef tallow or sucrose diets. (3) It did not significantly alter the total lipoprotein lipase activity in diaphragm, heart and adipose tissue. (4) The clearance of triacylglycerols from chylomicrons exhibited two t 1/2 values of about 0.6 and 6.9 min in rats fed the beef tallow diet. Benfluorex did not significantly alter these values. (5) Benfluorex did not significantly alter the rate of appearance of triacylglycerol in the blood of rats injected with Triton WR 1339 to block triacylglycerol uptake. It did, however, decrease the rise in circulating glucose which presumably resulted from the stress of the procedure. (6) Benfluorex decreased the extent and duration of the rise in serum corticosterone when rats maintained on the corn oil diet were fed acutely with fructose. It also decreased the circulating concentrations of glycerol, triacylglycerol and glucose after fructose feeding. (7) Rats fed on the corn oil diet and then treated with benfluorex had lower concentrations of circulating glucose, triacylglycerol, glycerol and fatty acids after being injected with 2-deoxyglucose. (8) It is proposed that some of the long-term hypoglycaemic and hypotriglyceridaemic effects of benfluorex could be mediated indirectly through changes in endocrine balance, perhaps via the serotonergic system and in particular, by decreasing the effects of stress hormones relative to insulin. The implications of these findings are discussed in relation to controlling metabolism in stress conditions and for the management of obesity, diabetes and atherosclerosis.