Uptake and attitudes regarding hepatitis A vaccine among childcare centre staff, administrators, and parents.

OBJECTIVES: Study objectives were to assess parents' and childcare agency staff's uptake of and attitudes/beliefs related to hepatitis A vaccine. STUDY DESIGN: Cross-sectional survey. METHODS: Questionnaires were administered to parents and staff in 23 St. Louis childcare agencies between September and December 2014. Categorical data were compared using Chi-squared test. Multivariate logistic regression, stratified by staff vs parents, was used to find predictors of hepatitis A vaccine uptake. RESULTS: In total, 351 staff and parents participated (response rate = 32%). More staff than parents had been offered or recommended to receive hepatitis A vaccine by a healthcare provider (55.4% vs 36.6% and 53.3% vs 28.7%, respectively; P = .001 for both). More staff than parents received hepatitis A vaccine (85.3% vs 67.5%, Chi-squared test = 11.0, P < .001). Predictors of staff vaccine uptake included being aware of CDC vaccination recommendations (OR = 11.2, CI = [1.4-91], P < .05), employer recommendation to get vaccinated (OR = 8.1, CI = [1.8-36.8], P < .01), and having a mandatory staff vaccination policy (OR = 4.8, CI = [1.2-19.7], P < .05). Predictors of parent vaccine uptake included being offered the vaccine by a healthcare provider (OR = 4.3, CI = [1.3-4.9], P < .001), being aware of the CDC vaccination recommendations (OR = 4.0, CI = [2.0-8.0], P < .001), and having received influenza vaccine previously (OR = 2.5, CI = [1.3-4.9], P < .01). CONCLUSION: In this study population, many childcare agency staff and parents have received hepatitis A vaccine, though staff immunization rates are at the minimum needed to reach herd immunity levels. Having employers encourage vaccination, offer free vaccine, or make vaccine available onsite could increase staff vaccination rates. Public health should partner with childcare agencies to increase staff vaccine uptake, which could result in community herd immunity.

Extrapulmonary tissue responses in cynomolgus macaques (Macaca fascicularis) infected with highly pathogenic avian influenza A (H5N1) virus.

The mechanisms responsible for virulence of influenza viruses in humans remain poorly understood. A prevailing hypothesis is that the highly pathogenic virus isolates cause a severe cytokinemia precipitating acute respiratory distress syndrome and multiple organ dysfunction syndrome. Cynomolgus macaques (Macaca fascicularis) infected with a human highly pathogenic avian influenza (HPAI) H5N1 virus isolate (A/Vietnam/1203/2004) or reassortants of human influenza virus A/Texas/36/91 (H1N1) containing genes from the 1918 pandemic influenza A (H1N1) virus developed severe pneumonia within 24 h postinfection. However, virus spread beyond the lungs was only detected in the H5N1 group, and signs of extrapulmonary tissue reactions, including microglia activation and sustained up-regulation of inflammatory markers, most notably hypoxia inducible factor-1alpha (HIF-1alpha), were largely limited to this group. Extrapulmonary pathology may thus contribute to the morbidities induced by H5N1 viruses.

Functional genomic and serological analysis of the protective immune response resulting from vaccination of macaques with an NS1-truncated influenza virus.

We are still inadequately prepared for an influenza pandemic due to the lack of a vaccine effective for subtypes to which the majority of the human population has no prior immunity and which could be produced rapidly in sufficient quantities. There is therefore an urgent need to investigate novel vaccination approaches. Using a combination of genomic and traditional tools, this study compares the protective efficacy in macaques of an intrarespiratory live influenza virus vaccine produced by truncating NS1 in the human influenza A/Texas/36/91 (H1N1) virus with that of a conventional vaccine based on formalin-killed whole virus. After homologous challenge, animals in the live-vaccine group had greatly reduced viral replication and pathology in lungs and reduced upper respiratory inflammation. They also had lesser induction of innate immune pathways in lungs and of interferon-sensitive genes in bronchial epithelium. This postchallenge response contrasted with that shortly after vaccination, when more expression of interferon-sensitive genes was observed in bronchial cells from the live-vaccine group. This suggested induction of a strong innate immune response shortly after vaccination with the NS1-truncated virus, followed by greater maturity of the postchallenge immune response, as demonstrated with robust influenza virus-specific CD4+ T-cell proliferation, immunoglobulin G production, and transcriptional induction of T- and B-cell pathways in lung tissue. In conclusion, a single respiratory tract inoculation with an NS1-truncated influenza virus was effective in protecting nonhuman primates from homologous challenge. This protection was achieved in the absence of significant or long-lasting adverse effects and through induction of a robust adaptive immune response.

Integrated molecular signature of disease: analysis of influenza virus-infected macaques through functional genomics and proteomics.

Recent outbreaks of avian influenza in humans have stressed the need for an improved nonhuman primate model of influenza pathogenesis. In order to further develop a macaque model, we expanded our previous in vivo genomics experiments with influenza virus-infected macaques by focusing on the innate immune response at day 2 postinoculation and on gene expression in affected lung tissue with viral genetic material present. Finally, we sought to identify signature genes for early infection in whole blood. For these purposes, we infected six pigtailed macaques (Macaca nemestrina) with reconstructed influenza A/Texas/36/91 virus and three control animals with a sham inoculate. We sacrificed one control and two experimental animals at days 2, 4, and 7 postinfection. Lung tissue was harvested for pathology, gene expression profiling, and proteomics. Blood was collected for genomics every other day from each animal until the experimental endpoint. Gross and microscopic pathology, immunohistochemistry, viral gene expression by arrays, and/or quantitative real-time reverse transcription-PCR confirmed successful yet mild infections in all experimental animals. Genomic experiments were performed using macaque-specific oligonucleotide arrays, and high-throughput proteomics revealed the host response to infection at the mRNA and protein levels. Our data showed dramatic differences in gene expression within regions in influenza virus-induced lesions based on the presence or absence of viral mRNA. We also identified genes tightly coregulated in peripheral white blood cells and in lung tissue at day 2 postinoculation. This latter finding opens the possibility of using gene expression arrays on whole blood to detect infection after exposure but prior to onset of symptoms or shedding.

Muscle glycogen depletion and subsequent replenishment affect anaerobic capacity of horses.

The purpose of this study was to determine the effect of muscle glycogen depletion and subsequent replenishment on anaerobic capacity of horses. In a blinded crossover study, seven fit horses performed glycogen-depleting exercise on two occasions. Horses were infused after glycogen-depleting exercise with either 6 g/kg body wt of glucose as a 13.5% solution in 0.9% NaCl (Glu) or with 0.9% NaCl (Sal) of equivalent volume. Subsequently, horses performed a high-speed exercise test (120% of maximal rate of oxygen consumption) to estimate maximum accumulated oxygen deficit. Replenishment of muscle glycogen was greater (P < 0.05) in Glu [from 24.7 +/- 7.2 (SE) to 116.5 +/- 7 mmol/kg wet wt before and after infusion, respectively] than in Sal (from 23.4 +/- 7.2 to 47.8 +/- 5.7 mmol/kg wet wt before and after infusion, respectively). Run time to fatigue during the high-speed exercise test (97.3 +/- 8.2 and 70.8 +/- 8.3 s, P < 0.05), maximal accumulated oxygen deficit (105.7 +/- 9.3 and 82.4 +/- 10.3 ml O(2) equivalent/kg, P < 0.05), and blood lactate concentration at the end of the high-speed exercise test (11.1 +/- 1.4 and 9.2 +/- 3.7 mmol/l, P < 0.05) were greater for Glu than for Sal, respectively. We concluded that decreased availability of skeletal muscle glycogen stores diminishes anaerobic power generation and capacity for high-intensity exercise in horses.

Rhesus monkey model for fetal gene transfer: studies with retroviral- based vector systems.

Many life-threatening conditions that can be diagnosed early in gestation may be treatable in utero using gene therapy. In order to determine in utero gene transfer efficiency and safety, studies were conducted with fetal rhesus monkeys as a model for the human. Included in these studies were Moloney murine leukemia virus (MLV)-based amphotropic retrovirus, vesicular stomatitis virus-G (VSV-G) pseudotyped MLV, and a VSV-G pseudotyped HIV-1-based vector, all expressing the enhanced green fluorescent protein (EGFP) as a reporter gene and driven by a cytomegalovirus-immediate early promoter (N = 16). Rhesus monkey fetuses were administered viral vector supernatant preparations by the intraperitoneal (ip) (N = 14) or intrahepatic (ih) (N = 2) routes via ultrasound guidance at 55 +/- 5 days gestation (late first trimester; term 165 +/- 10 days). Fetuses were monitored sonographically, specimens were collected prenatally and postnatally, and tissue harvests were performed at birth or 3 or 6 months postnatal age (3-10 months post-gene transfer). PCR analyses demonstrated that transduced cells were present at approximately 1.2% in peripheral blood mononuclear cells from fetuses administered amphotropic MLV, <0.5% in fetuses receiving MLV/VSV-G, and approximately 4.2% for the lentiviral vector, which decreased to 2% at birth. Hematopoietic progenitors showed that overall (mean of all time points assessed), approximately 25% of the collected colonies were positive for the EGFP transgene with the lentiviral vector, which was significantly greater than results achieved with the MLV-based vector systems (4-9%; P < or = 0.001-0.016). At necropsy, 0.001-10% of the total genomic DNA was positive for EGFP in most tissues for all groups. EGFP-positive fluorescent cells were found in cell suspensions of thymus, liver, spleen, lymph nodes, cerebral cortex, and bone marrow (0.5-6%). Overall, the results of these studies have shown: (1) healthy infants expressing vector sequences up to 10 months post-gene transfer, (2) fetal primate administration of retroviral vectors results in gene transfer to multiple organ systems, (3) the highest level of gene transfer to hematopoietic progenitors was observed with the lentiviral vector system, and (4) there was no evidence of transplacental transfer of vector sequences into the dams. The rhesus monkey is an important preclinical primate model system for exploring gene transfer approaches for future applications in humans.

Analysis of gene transfer efficiency of retrovirus producer cell transplantation for in situ gene transfer to hematopoietic cells.

OBJECTIVE: The aim of this study was to assess the gene transfer efficiency of an in situ administration protocol for hematopoietic stem/progenitor cells in the rhesus macaque (Macaca mulatta) animal model. MATERIALS AND METHODS: Moloney murine leukemia virus amphotropic vector producer cells (1--2 x 10(8) cells/animal) were transplanted into the femoral bone marrow cavities of six macaques. To determine if the levels of gene transfer could be increased, a second injection at the same dose of producer cells was performed into the iliac crest in three of the six macaques. RESULTS: We demonstrated that 0.02-0.1% of peripheral blood mononuclear cells contained the vector transgene for up to 12 months following the initial administration of producer cells. Hematopoietic progenitor cell assays indicated that the neomycin phosphotransferase gene was detected in 10--30% of progenitor cell colonies. A humoral immune response directed toward viral particles was demonstrated in all animals. Additionally, we demonstrated that an increase in the levels of transduced cells, up to 1% of circulating peripheral blood mononuclear cells and granulocytes, contain the transgene following producer cell readministration. CONCLUSIONS: These data demonstrate the successful in situ gene transfer to hematopoietic stem/progenitor cells and circulating peripheral blood mononuclear cells that persists as long as 12 months postinjection, in the absence of any preconditioning.

Effect of dietary supplements containing antioxidants on attenuation of muscle damage in exercising sled dogs.

OBJECTIVE: To determine whether dietary antioxidants would attenuate exercise-induced increases in plasma creatine kinase (CK) activity in sled dogs. ANIMALS: 41 trained adult sled dogs. PROCEDURE: Dogs, randomly assigned to 2 groups, received the same base diet throughout the study. After 8 weeks on that diet, 1 group (21 dogs) received a daily supplement containing vitamins E (457 U) and C (706 mg) and beta-carotene (5.1 mg), and a control group (20 dogs) received a supplement containing minimal amounts of antioxidants. After 3 weeks, both groups performed identical endurance exercise on each of 3 days. Blood samples were collected before and 3 weeks after addition of supplements and after each day of exercise. Plasma was analyzed for vitamins E and C, retinol, uric acid, triglyceride, and cholesterol concentrations, total antioxidant status (TAS), and CK activity. RESULTS: Feeding supplements containing antioxidants caused a significant increase in vitamin E concentration but did not change retinol or vitamin C concentrations orTAS. Exercise caused significantly higher CK activity, but did not cause a significant difference in CK activity between groups. Exercise was associated with significantly lower vitamin E, retinol, and cholesterol concentrations and TAS but significantly higher vitamin C, triglyceride, and uric acid concentrations in both groups. CONCLUSIONS AND CLINICAL RELEVANCE: Use of supplements containing the doses of antioxidants used here failed to attenuate exercise-induced increases in CK activity. Muscle damage in sled dogs, as measured by plasma CK activity, may be caused by a mechanism other than oxidant stress.

Factors influencing first remission and survival in 145 dogs with lymphoma: a retrospective study.

Records of 145 dogs diagnosed with lymphoma were reviewed to evaluate for factors influencing duration of remission and survival. Dogs with histories of certain chronic inflammatory diseases were 3.23 times more likely to relapse (relative risk, 3.23) than the overall population. Dogs with World Health Organization (WHO) stage IV lymphoma or those treated with a protocol containing cyclophosphamide, doxorubicin, vincristine, prednisone, and sulfatrimethoprim (CHOP) had lower relative risks of relapse (0.32 and 0.085, respectively). Progressive disease after induction, gastrointestinal toxicity from induction, and clinical signs (i.e., substage b lymphoma) were associated with higher relative risks of death (3.5, 2.64, and 2.02, respectively).

Effects of dietary antioxidant supplementation on oxidative damage and resistance to oxidative damage during prolonged exercise in sled dogs.

OBJECTIVES: To determine effects of dietary antioxidant supplementation on plasma concentrations of antioxidants, exercise-induced oxidative damage, and resistance to oxidative damage during exercise in Alaskan sled dogs. ANIMALS: 62 Alaskan sled dogs. PROCEDURE: Dogs were matched for age, sex, and ability and assigned to 1 of 3 groups: sedentary and nonsupplemented (control [C]; n = 21), exercised and supplemented (S; 22), and exercised and nonsupplemented (N; 19). Dogs in group S were given 400 units of alpha-tocopherol acetate, 3 mg of beta-carotene, and 20 mg of lutein orally per day for 1 month, then dogs in groups S and N completed 3 days of exercise. Blood samples were collected before and after 1 and 3 days of exercise and after 3 days of rest. Plasma antioxidant concentrations were determined, and oxidative damage to DNA (plasma 7,8 dihydro-8-oxo-2'deoxyguanosine [8-oxodG] concentration) and membrane lipids (plasma hydroperoxide concentration) and resistance of plasma lipoproteins to oxidation were assessed. RESULTS: Supplementation increased plasma concentrations of alpha-tocopherol, beta-carotene, and lutein. Plasma concentration of alpha-tocopherol increased and concentration of lutein decreased in group S with exercise. Concentration of 8-oxodG decreased in group S but increased in group N during and after exercise. Lag time of in vitro oxidation of lipoprotein particles increased with exercise in group S only. CONCLUSIONS AND CLINICAL RELEVANCE: Dietary supplementation with antioxidants resulted in increased plasma concentrations of antioxidants. Moreover, supplementation decreased DNA oxidation and increased resistance of lipoprotein particles to in vitro oxidation. Antioxidant supplementation of sled dogs may attenuate exercise-induced oxidative damage.