Characterization of Plaque-Sized Variants of Daniel's (DA) Strain in Theiler's Virus-Induced Epilepsy.

Epilepsy is a complex neurological disease characterized by recurrent seizures. Patients with viral encephalitis have a 16-fold increased risk of developing epilepsy, and this risk can persist for about 15 years after the occurrence of initial viral infection. Theiler's murine encephalomyelitis virus (TMEV) infection induces a well-characterized experimental model of epilepsy in C57BL/6 mice. In response to intracerebral (I.C.) injection of Daniel's (DA) strain of TMEV, there is vigorous immune response, which is detrimental to neurons and contributes to acute seizures, rendering mice susceptible to epilepsy. A comparative in vivo challenge study with either one of the two variants of the DA strain, small (DA-DS) or large (DA-CL) plaque forming variants, revealed differences in the diseases they induced in C57BL/6 mice. Compared to DA-CL-, DA-DS-infected mice exhibited significantly more seizures, higher clinical scores, neuroinflammation, and neuronal damage (mainly in the CA1-CA2 regions of hippocampus). Moreover, the brains of DA-DS infected mice contained approximately five-fold higher virus than those of DA-CL infected mice. A sequence comparison of the DA-CL and DA-DS genome sequences showed mutations in the leader (L) and L* proteins of DA-CL variant, which may be the cause of attenuating phenotype of DA-CL variant in the C57BL/6 mouse model of epilepsy.

Social disruption alters pain and cognition in an animal model of multiple sclerosis.

Although pain and cognitive deficits are widespread and debilitating symptoms of multiple sclerosis (MS), they remain poorly understood. Theiler's murine encephalomyelitis virus (TMEV) infection is an animal model of MS where disease course is exacerbated by prior stressors. Here chronic infection coupled with prior social stress increased pain behavior and impaired hippocampal-dependent memory consolidation during the demyelinating phase of disease in SJL mice. These results suggest that the TMEV model may be useful in investigating pain and cognitive impairments in MS. However, in contrast to prior Balb/cJ studies, stress failed to consistently alter behavioral and physiological indicators of disease course.

The assessment of bone mineral content and density of the lumbar spine and proximal femur in US submariners.

UNLABELLED: The submarine environment is unique in that there is limited space and no sunlight, which may negatively affect skeletal health and lead to accelerated bone loss, osteoporosis, and fractures. INTRODUCTION: The primary purpose of this study was to determine whether there was an association with submarine service, specifically time spent at sea, and bone mineral content (BMC) and bone mineral density (BMD) of the lumbar spine and dual proximal femur (total hip and femoral neck) measured by DXA. METHODS: This is a cross-sectional study of 462 submariners 20-91 years old. Variables included in the analysis were age, height, race, alcohol intake, tobacco use, fracture history, conditions, and medications known to cause bone loss and osteoporosis and submarine service. RESULTS: Of the submarine service predictors, only serving onboard a diesel submarine was determined to be independently associated with a reduction in BMD of the total hip and femur neck, while no submarine service predictor increased the odds of having low BMD. In submariners 50+ years old, the age-adjusted prevalence of osteopenia was 15.7 % (lumbar spine) and 40.4 % (femur neck), while the prevalence of osteoporosis was 4.8 % (lumbar spine) and 4.2 % (femur neck), rates that did not differ from NHANES 2005-2008. In submariners <50 years old, 3.1 % was below the expected range for age. The proportion of submariners 50+ years old that met the FRAX criteria for pharmacological treatment was 12 %. CONCLUSIONS: Intermittent periods of submergence that can range from a few days to 3+ months do not appear to compromise skeletal health differently than the general population.

Neonatal experience interacts with adult social stress to alter acute and chronic Theiler's virus infection.

Previous research has shown that neonatal handling has prolonged protective effects associated with stress resilience and aging, yet little is known about its effect on stress-induced modulation of infectious disease. We have previously demonstrated that social disruption stress exacerbates the acute and chronic phases of the disease when applied prior to Theiler's virus infection (PRE-SDR) whereas it attenuates disease severity when applied concurrently with infection (CON-SDR). Here, we asked whether neonatal handling would protect adult mice from the detrimental effects of PRE-SDR and attenuate the protective effects of CON-SDR on Theiler's virus infection. As expected, handling alone decreased IL-6 and corticosterone levels, protected the non-stressed adult mice from motor impairment throughout infection and reduced antibodies to myelin components (PLP, MBP) during the autoimmune phase of disease. In contrast, neonatal handling X PRE/CON-SDR elevated IL-6 and reduced corticosterone as well as increased motor impairment during the acute phase of the infection. Neonatal handling X PRE/CON-SDR continued to exacerbate motor impairment during the chronic phase, whereas only neonatal handling X PRE-SDR increased in antibodies to PLP, MOG, MBP and TMEV. Together, these results imply that while handling reduced the severity of later Theiler's virus infection in non-stressed mice, brief handling may not be protective when paired with later social stress.

Directional dispersal between mid-ocean ridges: deep-ocean circulation and gene flow in Ridgeia piscesae.

This study examined relationships between bathymetrically induced deep-ocean currents and the dispersal of the hydrothermal vent tubeworm Ridgeia piscesae along the northeast Pacific ridge system. A robust diagnostic model of deep-ocean circulation in this region predicted strong southeasterly currents following contours of the Blanco Transform Fault, a 450-km lateral offset that separates the Gorda and Juan de Fuca ridge systems. Such currents should facilitate the southward dispersal of R. piscesae larvae. Immigration rates for populations north and south of the Blanco Transform Fault were estimated from molecular population genetic data. Mitochondrial DNA evidence revealed population subdivision across the Blanco Transform Fault, and a strong directional bias in gene flow that was consistent with predictions of the circulation model. The distribution of mitochondrial diversity between the northern and southern populations of R. piscesae suggests that the Gorda Ridge tubeworms have maintained larger effective population sizes than the northern populations, a pattern that also exists in co-occurring limpets. Together, these data suggest that the northern vent fields may experience a higher frequency of habitat turnover and consequently more rapid losses of genetic diversity.

Inter- and intra-observer reliability of radiographic scores commonly used for the evaluation of haemophilic arthropathy.

Although the severity of haemophilic arthropathy is commonly assessed using established radiographic scoring systems, there is limited available information about their inter- and intra-observer reliability. The purpose of the present study was to establish the inter-observer reliability (IEOR) and intra-observer reliability (IAOR) of three different methods available for the classification of haemophilic arthropathy, including the Arnold and Hilgartner classification, a modification to the Arnold and Hilgartner system described by Luck et al., and the classification described by Pettersson et al. Antero-posterior and lateral radiographs of 54 haemophilic joints were included for the analysis. To determine the IEOR for each one of the three radiographic systems, the radiographs were randomly evaluated by four observers, including two orthopaedic surgeons, one orthopaedic resident and one haematologist. For the determination of IAOR, all four reviewers repeated the assessment in a similar fashion, after a period of at least 2 weeks. IEOR and IAOR for the three classification systems was established using kappa (kappa) statistics. A Spearman rank correlation was used to determine the similarities between each reviewer's own interpretative scales. The IEOR was low for the Arnold and Hilgartner system (kappa = 0.35, P < or = 0.001) and the Luck system (kappa = 0.38, P < or = 0.001), but even lower for the Pettersson system (kappa = 0.06, P = 0.1). For the Pettersson system, particularly low kappa values were observed for the presence or absence of osteoporosis (kappa = 0.11, P = 0.0027), enlarged epiphysis (kappa = 0.10, P = 0.0039), erosion of joint margins (kappa = 0.11, P = 0.0018), and joint deformity (kappa = 0.16, P = 0.00001). However, a relatively high Spearman rank correlation for all three scales [r(s) = 0.75 (P < 0.001) for Arnold and Hilgartner system, r(s) = 0.74 (P < 0.001) for the Luck system and r(s) = 0.81 (P < 0.001) for Pettersson system] indicated an overall, general agreement among the reviewers with regard to the severity of the haemophilic arthropathy. There was a moderate IAOR value for both, the Arnold and Hilgartner system (kappa = 0.57, P = 0.00001) and the Luck system (kappa = 0.62, P = 0.00001) with a low IAOR value for the Pettersson system [kappa = 0.22, P = 0.00001). Currently available radiographic scoring systems for haemophilic arthropathy have low inter- and intra-observer reliability rates. Improvements, either through education or modification of the scoring systems, are critical in an era where correlations between clinical and radiographic scores have received significant attention.

A comparison of the neurotropism of Theiler's virus and poliovirus in CBA mice.

Theiler's murine encephalomyelitis virus (TMEV) and poliovirus infect the central nervous system (CNS) and cause neurological damage. The exact route by which TMEV and polioviruses enter the CNS remains, for the most part, unknown, although the neural and/or the hematogenous pathway have both been postulated. To explore these hypotheses, this research focuses on both the site of entry and the pathway used to invade the CNS. Following different inoculation sites of the GDVII strain of Theiler's virus or Lansing Type 2 poliovirus in CBA mice, the incidence of paralysis and/or encephalitis was evaluated on the basis of clinical signs and histopathology. The forms of paralysis displayed corresponded to the site of viral inoculation. Following intramuscular (i.m.), intraperitoneal (i.p.), and footpad routes of injection, bilateral and or contralateral paralyses were observed for both TMEV and poliovirus. In mice injected intratongue and in the hypoglossal nerve, tongue paralysis or paralysis of the forelimb, which progressed to bilateral forelimb paralysis, was observed, additionally the penis of most infected males was protruded. Intracranial (i.c.) injections with type II poliovirus strain resulted in forelimb paralysis. Intravenous (i.v.), injections with TMEV also resulted in forelimb paralysis. Thus Lansing Type II poliovirus and TMEV infections of CBA mice, result in similar incidence of paralysis and histopathological findings.

Restraint stress decreases virus-induced pro-inflammatory cytokine mRNA expression during acute Theiler's virus infection.

Stressful life events have been associated with the onset and/or exacerbation of multiple sclerosis (MS). Our previous studies have indicated that restraint stress (RS) reduces inflammation and virus-induced chemokine expression in the Theiler's virus-induced demyelination (TVID) model of MS. Here we report that RS significantly reduced the virus-induced interferon-gamma mRNA levels in the brain. Additionally, mRNA levels of lymphotoxin-beta, tumor necrosis factor-alpha, and interferon-gamma in the brain were negatively correlated with viral titers in the brain. These results indicated an immunosuppressive effect of stress during early TVID causing impaired viral clearance, which may be a potential exacerbating factor for later demyelination.

Complications of reperfusion in acute aortic artery occlusion following saddle embolization originating from an atrial myxoma.

A 58-year-old man presented to the hospital with an 8-hour history of acute-onset bilateral lower limb ischemia. A large saddle embolus had occluded the aorta and could not be removed by balloon endarterectomy through the femoral arteries. Successful open aortic and femoral thromboembolectomy followed by extensive fasciotomies was accompanied by severe reperfusion injury. Life-threatening hyperkalemia was associated with three episodes of intraoperative ventricular fibrillation and ventricular tachycardia requiring cardiac massage and defibrillation. A dextrose-insulin-bicarbonate infusion was required to correct the hyperkalemia. Rhabdomyolysis developed at 24 hours, causing marked myoglobinuria and acute renal failure, which required hemofiltration. Histology of the recovered embolus confirmed an atrial myxoma, and when the patient had fully recovered, open cardiac surgery was carried out to resect the tiny stump of residual myxoma. Rhabdomyolysis associated with a myxomatous saddle embolus has not been previously reported. This case highlights the need for pre- and perioperative measures to be taken to overcome hyperkalemia and acute renal failure when revascularizing acute, massive, prolonged ischemia of the lower body.

Properties of drug-containing spherical pellets produced by a hot-melt extrusion and spheronization process.

The objectives of this study were to investigate the particle size distribution, morphology and dissolution properties of spherical pellets produced by hot-melt extrusion and spheronization and to compare the properties of hot-melt extruded pellets with beads manufactured by a traditional wet-mass extrusion and spheronization method. Spherical pellets were produced by hot-melt extrusion without the use of water or other solvents. A powder blend of theophylline, Eudragit Preparation 4135 F, microcrystalline cellulose and polyethylene glycol 8000 was hot melt-extruded and the resulting composite rod was cut into cylindrical pellets. The pellets were then spheronized in a traditional spheronizer at an elevated temperature. The same powder blend was processed using conventional wet-mass techniques. Unlike wet-mass extruded pellets, pellets prepared from hot-melt extrusion displayed both a narrow particle size distribution and controlled drug release in dissolution media less than pH 7.4. Scanning electron microscopy, X-ray diffraction and porosity measurements were employed to explain the differences in drug release rates of theophylline from pellets produced by the two processing techniques. Theophylline release from the hot-melt extruded pellets was described using the Higuchi diffusion model, and drug release rates from wet-granulated and melt-extruded pellets did not change after post-processing thermal treatment.

Dispersal barriers and isolation among deep-sea mussel populations (Mytilidae: Bathymodiolus) from eastern Pacific hydrothermal vents.

Deep-sea hydrothermal vent species are widely dispersed among habitat islands found along the global mid-ocean ridge system. We examine factors that affect population structure, gene flow and isolation in vent-endemic mussels of the genus Bathymodiolus from the eastern Pacific Ocean. Mussels were sampled from localities including the Galapagos Rift (GAR, 0 degrees 48' N; 86 degrees 10' W) and the East Pacific Rise (EPR, 13 degrees N to 32 degrees S latitude) across a maximum distance of 4900 km. The sampled range crossed a series of topographical features that interrupt linear aspects of the ridge system, and it encompassed regions of strong cross-axis currents that could impede along-axis dispersal of mussel larvae. Examinations of mitochondrial DNA sequences and allozyme variation revealed significant barriers to gene flow along the ridge axis. All populations from the GAR and EPR from 13 degrees N to 11 degrees S were homogeneous genetically and appeared to experience unimpeded high levels of interpopulational gene flow. In contrast, mussels from north and south of the Easter Microplate were highly divergent (4.4%), possibly comprising sister-species that diverged after formation of the microplate approximately 4.5 Ma. Strong cross-axis currents associated with inflated bathymetry of the microplate region may reinforce isolation across this region.

Production, characterization, and cross-reactivity studies of monoclonal antibodies against the coccidiostat nicarbazin.

A cELISA was developed for the coccidiostat nicarbazin. On the basis of previous computer-assisted molecular modeling studies, p-nitrosuccinanilic acid (PNA-S) was selected as a hapten to produce antibodies to 4,4'-dinitrocarbanilide (DNC), the active component of the coccidiostat nicarbazin. Synthesis is described for the hapten [p-nitro-cis-1,2-cyclohexanedicarboxanilic acid (PNA-C)] used in a BSA conjugate as a plate coating antigen. Monoclonal antibodies (Mabs) were isolated that compete with nicarbazin, having IgM(kappa) isotype. Because of the lack of water solubility of nicarbazin, N,N-dimethylformamide (DMF) (3%, v/v) and acetonitrile (ACN) (10%, v/v) were added to the assay buffer to achieve solubility of nicarbazin and related compounds. The Nic 6 Mabs had an IC(35) value for nicarbazin of 0.92 nmol/mL, with a limit of detection of 0.33 nmol/mL. Nic 6 exhibited high cross-reactivity for PNA-S and PNA-C, and 3-nitrophenol, 4-nitrophenol, and 1-(4-chlorophenyl)-3-(4-nitrophenyl) urea. However, Nic 6 had little or no cross-reactivity with 15 other related compounds.

Role of Campylobacter jejuni potential virulence genes in cecal colonization.

Campylobacter jejuni, a common commensal in chickens, is one of the leading causes of bacterial gastroenteritis in humans worldwide. The aims of this investigation were twofold. First, we sought to determine whether mutations in the C. jejuni ciaB and pldA virulence-associated genes impaired the organism's ability to colonize chickens. Second, we sought to determine if inoculation of chicks with C. jejuni mutants could confer protection from subsequent challenge with the C. jejuni wild-type strain. The C. jejuni ciaB gene encodes a secreted protein necessary for the maximal invasion of C. jejuni into cultured epithelial cells, and the pldA gene encodes a protein with phospholipase activity. Also included in this study were two additional C. jejuni mutants, one harboring a mutation in cadF and the other in dnaJ, with which we have previously performed colonization studies. In contrast to results with the parental C. jejuni strain, viable organisms were not recovered from any of the chicks inoculated with the C. jejuni mutants. To determine if chicks inoculated with the C. jejuni mutants become resistant to colonization by the C. jejuni parental strain upon subsequent challenge, chicks were inoculated either intraperitoneally (i.p.) or both orally and i.p. with the C. jejuni mutants. Inoculated birds were then orally challenged with the parental strain. Inoculation with the C. jejuni mutants did not provide protection from subsequent challenge with the wild-type strain. In addition, neither the C. jejuni parental nor the mutant strains caused any apparent morbidity or mortality of the chicks. We conclude that mutations in genes cadF, dnaJ, pldA, and ciaB impair the ability of C. jejuni to colonize the cecum, that chicks tolerate massive inoculation with these mutant strains, and that such inoculations do not provide biologically significant protection against colonization by the parental strain.

Effects of tannic acid on cecal volatile fatty acids and susceptibility to Salmonella typhimurium colonization in broiler chicks.

Young chickens are more susceptible to Salmonella colonization than older chickens that have developed resistance with age as native microflora become established. Elevated concentrations of cecal propionic acid and total volatile fatty acids (VFA) have been observed by many researchers to be indicators of establishment of anaerobic microflora and protection against Salmonella colonization of the ceca. Disruption of the native microflora or competitive exclusion (CE) cultures by components of diets, such as tannic acid (TA), could alter the concentrations of propionic acid and total VFA and possibly affect Salmonella colonization. Two experiments were conducted using day-of-hatch, mixed-sex broiler chicks to evaluate the effects of TA on cecal VFA and the susceptibility to Salmonella colonization. All chicks in both experiments were challenged orally with 10(4) cfu of Salmonella typhimurium (ST) on Day 3 (Experiment 1) or Day 4 (Experiment 2). One-half of the chicks were orally gavaged on the day of hatch with a CE culture (PREEMPT) and were fed diets containing 0, 0.75, or 1.5% TA for up to 12 d of age. Chicks were maintained in batteries in separate rooms for the experimental period. There were some alterations in concentrations of cecal propionic acid or total VFA in chicks fed diets containing 0.75 or 1.5% TA in non CE-treated chicks and in CE-treated chicks. No significant differences were observed for numbers of Salmonella cecal culture-positive chicks or in the numbers of ST in the cecal contents due to dietary content of TA. With minor exceptions, the chicks treated with the CE culture had higher cecal concentrations of propionic acid and were less susceptible to Salmonella colonization than the non CE-treated chicks. Further research is necessary to determine the biological significance of these changes.

Biogeography and ecological setting of Indian Ocean hydrothermal vents.

Within the endemic invertebrate faunas of hydrothermal vents, five biogeographic provinces are recognized. Invertebrates at two Indian Ocean vent fields (Kairei and Edmond) belong to a sixth province, despite ecological settings and invertebrate-bacterial symbioses similar to those of both western Pacific and Atlantic vents. Most organisms found at these Indian Ocean vent fields have evolutionary affinities with western Pacific vent faunas, but a shrimp that ecologically dominates Indian Ocean vents closely resembles its Mid-Atlantic counterpart. These findings contribute to a global assessment of the biogeography of chemosynthetic faunas and indicate that the Indian Ocean vent community follows asymmetric assembly rules biased toward Pacific evolutionary alliances.

Effects of feed withdrawal and transport on cecal environment and Campylobacter concentrations in a swine surgical model.

The objective of the present study was to evaluate how feed withdrawal and transportation influenced the cecal environment and cecal populations of Campylobacter in swine. Four miniature Yucatan gilts (8.8 kg), naturally infected with Campylobacter jejuni, were surgically implanted with cecal cannulas. The gilts were fasted for 48 h. Samples of cecal contents were collected for 7 days prior to and for 7 days after the fast, and mean values were determined for pH, volatile fatty acids (VFA), and CFU enumeration of C. jejuni. This was replicated three times. In another trial, gilts (full-fed) were transported in a livestock trailer for 4 h and cecal samples were collected before and after transport and analyzed for pH, VFA, and CFU. Following a 48-h fast, cecal pH increased (P < 0.05) by 1 unit; acetic and propionic acids decreased (P < 0.05) by 61% and 71%, respectively; and there was a twofold log10 increase (P < 0.05) in CFU/g cecal content of C. jejuni. Values of pH, VFA, and CFU of C. jejuni did not change in cecal samples from gilts following transportation. These data are important for food safety considerations because feed withdrawal, commonly associated with shipping and slaughter, can increase Campylobacter concentrations in the pig intestinal tract.

Cecal volatile fatty acids and broiler chick susceptibility to Salmonella typhimurium colonization as affected by aflatoxins and T-2 toxin.

Four experiments were conducted using day-of-hatch, mixed-sex broiler chicks to evaluate the effects of aflatoxins and T-2 toxin on cecal volatile fatty acids (VFA) and the susceptibility to Salmonella colonization. All chicks in these experiments were challenged orally with 10(4) cfu of Salmonella typhimurium (ST) on Day 3. In Experiments 1 and 2, chicks were fed diets containing 0, 2.5, or 7.5 mg aflatoxins/kg of diet and were allowed to develop their microflora naturally. In Experiment 3, all chicks were orally gavaged on the day of hatch with a competitive exclusion (CE) culture (PREEMPT) and were fed diets containing 0, 2.5, or 7.5 mg T-2 toxin/kg. In Experiment 4, the chicks were fed diets containing 0, 7.5, or 15.0 mg T-2 toxin/kg and one-half of the chicks were orally gavaged on the day of hatch with the CE culture. In Experiments 1 and 2, with the exception of increased total VFA at 5 d in chicks fed the 7.5 mg T-2 aflatoxins/kg diet, there were no treatment effects on cecal propionic acid, total VFA, or incidence or severity of ST colonization. In Experiment 3, the only alteration in concentration of cecal propionic acid or total VFA was a significant reduction in total VFA at 5 d in chicks fed the 2.5 mg T-2 toxin/kg diet. No significant treatment differences were observed for numbers of Salmonella cecal culture-positive chicks or for numbers of ST in the cecal contents. In Experiment 4, with minor exceptions, the chicks treated with the CE culture had higher cecal concentrations of propionic acid and were less susceptible to Salmonella colonization than the non-CE-treated chicks. In the non-CE-treated chicks, T-2 toxin had no effect on any of the parameters, and 85 to 90% of the chicks were Salmonella cecal culture-positive. In the CE-treated chicks, there was a decrease in propionic acid concentration at 3 and 11 d and an increase in susceptibility to Salmonella colonization of the chicks fed the 15.0 mg T-2 toxin/kg diet. These results indicate that cecal concentrations of VFA can be affected by toxins, such as high concentrations of T-2 toxin, and that resistance to Salmonella colonization may be reduced. Further research is necessary to determine the biological significance of these changes.

Anti-inflammatory effects of ergotamine in steers.

The objective of this experiment was to investigate whether the ergot alkaloid, ergotamine (ET), an alkaloid used to model fescue toxicosis in cattle, modifies the response of cattle to endotoxin (LPS) challenge. Steers (n = 16) were divided into the following treatment groups: control (C), ergotamine (ET), endotoxin (LPS), and ET + LPS. ET and ET + LPS groups received a single bolus intravenous injection of ET (40 microg. kg. body wt(-1)), whereas C and LPS steers received a single bolus injection of sterile vehicle. Thirty minutes after ET/vehicle administration, a single bolus intravenous injection of LPS (0.2 microg. kg. body wt(-1)) was given. Blood was collected at various time points for 48 hr post. Endotoxin increased rectal temperature (RT) and the circulating levels of tumor necrosis factor-alpha (TNF-alpha), cortisol, haptoglobin (Hp), thromboxane B(2) (TXB(2)). The circulating Hp, TNF-alpha, and TXB(2) increases were blunted by pretreatment with ET compared with ET + LPS. Ergotamine by itself increased circulating cortisol and RT, whereas it decreased serum prolactin (PRL). Therefore, whereas administration of LPS at 0.2 microg/kg to steers resulted in an expected response, the combination of ET + LPS attenuated major effects of LPS alone. Thus, acute administration of ET appeared to be anti-inflammatory as it decreased the inflammatory response to LPS, an effect likely driven at least in part by the ET-caused cortisol increase.