Effects of Mating and Social Exposure on Cell Proliferation in the Adult Male Prairie Vole (Microtus ochrogaster).

Microtus ochrogaster is a rodent with a monogamous reproductive strategy characterized by strong pair bond formation after 6 h of mating. Here, we determine whether mating-induced pair bonding increases cell proliferation in the subventricular zone (SVZ), rostral migratory stream (RMS), and dentate gyrus (DG) of the hippocampus in male voles. Males were assigned to one of the four groups: (1) control: males were placed alone in a clean cage; (2) social exposure to a female (SE m/f): males that could see, hear, and smell a sexually receptive female but where physical contact was not possible, because the animals were separated by an acrylic screen with small holes; (3) social exposure to a male (SE m/m): same as group 2 but males were exposed to another male without physical contact; and (4) social cohabitation with mating (SCM): males that mated freely with a receptive female for 6 h. This procedure leads to pair bond formation. Groups 2 and 3 were controls for social interaction. Male prairie voles were injected with 5-bromo-2'-deoxyuridine (BrdU) during the behavioral tests and were sacrificed 48 h later. Brains were processed to identify the new cells (BrdU-positive) and neuron precursor cells (neuroblasts). Our principal findings are that in the dorsal region of the SVZ, SCM and SE m/f and m/m increase the percentage of neuron precursor cells. In the anterior region of the RMS, SE m/f decreases the percentage of neuron precursor cells, and in the medial region SE m/f and m/m decrease the number of new cells and neuron precursor cells. In the infrapyramidal blade of the subgranular zone of the DG, SE m/m and SCM increase the number of new neuron precursor cells and SE m/m increases the percentage of these neurons. Our data suggests that social interaction, as well as sexual stimulation, leads to pair bonding in male voles modulating cell proliferation and differentiation to neuronal precursor cells at the SVZ, RMS, and DG.

[Long-term individual cognitive stimulation program in patients with mild neurocognitive disorder: a pilot study]. / Programa de estimulacion cognitiva individual de larga duracion para personas con trastorno neurocognitivo leve: estudio piloto.

INTRODUCTION: There is evidence to suggest that cognitive stimulation produces cognitive benefits in people with mild neurocognitive disorder. However, the effect has been previously demonstrated to be minimal to moderate and the effect of long-term individual interventions, namely on specific cognitive domains, is unknown. AIM: To assess the efficacy, feasibility and acceptability of a long-term individual cognitive stimulation intervention for patients with mild neurocognitive disorder. PATIENTS AND METHODS: Patients (n = 30) with mild neurocognitive disorder were assigned to a cognitive stimulation intervention group (n = 15) or to a control group (n = 15). The intervention consisted of 88 individual sessions, approximately 45 minutes long, with two sessions per week. External evaluators assessed the level of alteration in cognitive performance, depressive symptoms and the level of independence in the performance of basic activities of daily living. RESULTS: After the intervention, a significant improvement was found in the intervention group compared to the control group in overall cognitive performance (d = 0.83), specifically in the language domain (d until 1.50). There were also lower depressive symptoms in the intervention group compared to the control group (d = 0.93). Only 6.7% of the participants dropped out the study, with participants attending a mean of 83 ± 12.1 sessions. CONCLUSIONS: The results support the efficacy, feasibility and acceptability of the intervention for mild neurocognitive disorder and justify a randomized controlled trial of the program with a larger sample.

TITLE: Programa de estimulacion cognitiva individual de larga duracion para personas con trastorno neurocognitivo leve: estudio piloto.Introduccion. Existen evidencias que sugieren que la estimulacion cognitiva produce beneficios cognitivos en personas con trastorno neurocognitivo leve. Sin embargo, el tamaño del efecto encontrado es de pequeño a moderado, y se desconoce el efecto de las intervenciones individuales de larga duracion y, mas concretamente, sobre dominios cognitivos especificos. Objetivo. Evaluar la eficacia, viabilidad y aceptabilidad de una intervencion de estimulacion cognitiva individual de larga duracion para personas con trastorno neurocognitivo leve. Pacientes y metodos. Un total de 30 personas con trastorno neurocognitivo leve fueron asignadas a un grupo de intervencion de estimulacion cognitiva (n = 15) o a un grupo control (n = 15). La intervencion consistio en 88 sesiones individuales de unos 45 minutos, con una periodicidad de dos veces por semana. Evaluadores independientes valoraron el nivel de rendimiento cognitivo, los sintomas depresivos y el nivel de autonomia en la realizacion de actividades basicas de la vida diaria. Resultados. Tras la intervencion, se encontro una mejoria significativa en el grupo de intervencion en comparacion con el grupo control en el rendimiento cognitivo global (d = 0,83), concretamente en el dominio del lenguaje (d hasta 1,50), y una menor sintomatologia depresiva en el grupo de intervencion en comparacion con el control (d = 0,93). Solo un 6,7% de los participantes abandono el estudio, asistiendo a un promedio de 83 ± 12,1 sesiones. Conclusiones. Los resultados apoyan la eficacia, viabilidad y aceptabilidad de la intervencion, y justifican la realizacion de un ensayo controlado aleatorizado aplicado a una muestra mayor.

Assessing cellular DNA damage from a helium plasma needle.

The aim of the present study is to determine the deoxyribonucleic acid (DNA) damage by cells exposed to atmospheric pressure non-thermal plasma (APNTP). Mouse leukocytes embedded in agarose were exposed to the plasma at two different distances from a helium plasma needle outlet and during three different exposure periods. Damage was assessed by the single cell gel electrophoresis assay. The results indicate that, at 0.1 cm from the plasma needle, the exposure caused complete DNA fragmentation determined by the presence of so called "clouds". Samples exposed at 0.5 cm from the slide sample surface presented damage proportional to the exposure periods in terms of tail intensity, tail moment and "clouds" frequency. Studies performed with alkaline single cell gel electrophoresis assay to determine DNA breaks and alkali-labile sites, indicates that DNA damage produced by exposure to APNTP was caused mainly by oxidative radicals, rather than by UV light which causes cyclobutane pyrimidine dimers. These results allow us to conclude that plasma needle induced DNA breaks in mice leukocytes proportionally to exposure time.

Inactivation of Escherichia coli in water by pulsed dielectric barrier discharge in coaxial reactor.

An experimental study of ATCC (American Type Culture Collection) 8739 Escherichia coli bacteria inactivation in water by means of pulsed dielectric barrier discharge (PDBD) atmospheric pressure plasmas is presented. Plasma is generated by an adjustable power source capable of supplying high voltage 25 kV pulses, ∼30 µs long and at a 500 Hz frequency. The process was conducted in a ∼152 cm(3) cylindrical stainless steel coaxial reactor, endowed with a straight central electrode and a gas inlet. The bacterial concentration in water was varied from 10(3) up to 10(8) E. coli cells per millilitre. The inactivation was achieved without gas flow in the order of 82% at 10(8) colony-forming units per millilitre (CFU mL(-1)) concentrations in 600 s. In addition, oxygen was added to the gas supply in order to increase the ozone content in the process, raising the inactivation percentage to the order of 90% in the same treatment time. In order to reach a higher efficiency however, oxygen injection modulation is applied, leading to inactivation percentages above 99.99%. These results are similarly valid for lower bacterial concentrations.

The role of endoscopic extraperitoneal herniorrhaphy: where do we stand in 2005?

Inguinal hernia repair is a common surgical procedure, but the most effective surgical technique remains controversial. The evolution of laparoscopic techniques has allowed reproduction of open preperitoneal repair via an endoscopic total extraperitoneal (TEP) approach. More recently, the advent of comprehensive training in laparoscopy has allowed TEP to continue evolving as the feasibility of this approach gains recognition as a preferable technique. Once considered very difficult to learn, TEP currently is adequately taught in many surgical training programs. This report reviews the fundamentals and details various modifications that make this procedure more desirable than open procedures and other laparoscopic techniques. A resultant decrease in operative time, cost of the procedure, and morbidity to the patient is routine. In addition, the authors review their institutional experience and examine other current evidence-based data.

Survival of avian influenza virus H7N2 in SPF chickens and their environments.

The survival or clearance of the avian influenza virus (AIV) of subtype H7N2 in its chicken host was evaluated using experimentally infected specific pathogen free (SPF) chickens of different age groups. Birds of different ages were successfully infected with infectious doses ranging between 10(4.7) and 10(5.7) ELD50 per bird. In infected birds, the infective virus was undetectable usually by the third week following exposure. The infectivity or inactivation time of the H7N2 AIV in various environmental conditions was studied using chicken manure, heat, ethanol, pH, and disinfectants. The H7N2 AIV was effectively inactivated by field chicken manure in less than a week at an ambient temperature of 15-20 degrees C. At a pH 2, heating at 56 degrees C, and exposure to 70% ethanol or a specific disinfectant, the AIV infectivity was destroyed in less than 30 min.

Epidemiology, production losses, and control measures associated with an outbreak of avian influenza subtype H7N2 in Pennsylvania (1996-98).

An outbreak of H7N2 low-pathogenicity (LP) avian influenza (AI) occurred in a two-county area in Pennsylvania from December of 1996 through April of 1998. The outbreak resulted in infection of 2,623,116 commercial birds on 25 premises encompassing 47 flocks. Twenty-one (one premise with infection twice) of the twenty-five infected premises housed egg-laying chickens and one premise each had turkeys, layer pullets, quail, and a mixed backyard dealer flock. Despite dose proximity of infected flocks to commercial broiler flocks, no infected broilers were identified. Experimentally, when market age broilers were placed on an influenza-infected premise they seroconverted and developed oviduct lesions. The outbreak was believed to have originated from two separate introductions into commercial layer flocks from premises and by individuals dealing in sales of live fowl in the metropolitan New York and New Jersey live-bird markets. Source flocks for these markets are primarily in the northeast and mid-Atlantic areas, including Pennsylvania. Mixed fowl sold include ducks, geese, guinea hens, quail, chukar partridges, and a variety of chickens grown on perhaps hundreds of small farms. Infections with the H7N2 AI virus were associated with variable morbidity and temporary decreases in egg production ranging from 1.6% to 29.1% in commercial egg-laying chickens. Egg production losses averaged 4.0 weeks duration. Mortality ranged from 1.5 to 18.3 times normal (mean of 4.3 times normal). Duration of mortality ranged from 2 to 13 weeks (average of 3.9 weeks) in flocks not depopulated. Lesions observed were primarily oviducts filled with a mucous and white gelatinous exudates and atypical egg yolk peritonitis. Quarantine of premises and complete depopulation were the early measures employed in control of this outbreak. Epidemiological studies suggested that depopulation furthered the spread of influenza to nearby flocks. Thereafter, later control measures included quarantine, strict biosecurity, and controlled marketing of products.

Single-tube single-enzyme reverse transcriptase PCR assay for detection of bovine viral diarrhea virus in pooled bovine serum.

A reverse transcriptase PCR (RT-PCR) was developed for use as a diagnostic screening test for the detection of bovine viral diarrhea virus (BVDV) in pooled bovine serum samples. Individual serum samples from 60 dairy cattle herds located in Pennsylvania were evaluated by the microplate virus isolation method, and pooled sera were analyzed by RT-PCR. RT-PCR was sensitive and specific and detected a single viremic serum sample in up to 100 pooled serum samples. RT-PCR analysis of pooled sera provides a rapid and cost-effective method for the screening of cattle herds for the presence of animals persistently infected with BVDV.

Production and evaluation criteria of specific monoclonal antibodies to the hemagglutinin of the H7N2 subtype of avian influenza virus.

To enhance the rapidity in diagnosing the spread of avian influenza virus (AIV) in chicken layer flocks, studies were initiated to develop more sensitive and specific immunological and molecular methods for the detection of AIV. In this study, the purification of the hemagglutinin protein (H) from field isolates of H7N2, the production of monoclonal antibodies (MAbs), and their evaluation as diagnostic reagents are reported. Hybridomas were generated by fusion of SP2/0-Ag14 myelomas and spleen cells from immunized mice. Hybridomas secreting antibodies specific for the H protein were assayed by an ELISA and cloned using limiting dilution. The MAbs produced were characterized by hemagglutination inhibition (HI), immunohistochemistry (IHC), indirect fluorescent antibody assay (IFA), Western blots, and IFA flow cytometry using various AIV subtypes (i.e., H4N2, H5N3, H7N2). Of the various MAbs assayed, 6 had consistent and reproducible results in each of the assays used. The results obtained in this investigation enhanced the usage of the MAbs to viral H protein in the surveillance of AIV in chickens.

A gene for isolated congenital ptosis maps to a 3-cM region within 1p32-p34.1.

Hereditary isolated congenital ptosis is an autosomal dominant disorder with incomplete penetrance characterized by a variable degree of unilateral or bilateral drooping of the upper eyelids. We report linkage of this disorder in a large family to markers on chromosome 1p. In our sample of 37 meioses, nine informative markers did not recombine with the disease. D1S2677 gave a maximum two-point LOD score of 8.8 on the assumption of 90% penetrance (theta = 0). D1S447/2733 and D1S1616 flank the disease locus, with two-point LOD scores of 5.6/6.6 (theta = .04) and 4.9 (theta = .05), respectively, defining a region of 2.8 cM. FISH of YACs containing flanking recombinant markers localizes the gene to chromosome 1p32-p34.1. These data establish a map location for an isolated congenital ptosis gene and demonstrate that this disorder is genetically distinct from other extraocular muscle-specific disorders such as congenital fibrosis of the extraocular muscles and blepharophimosis.

Experimental reproduction of hypoglycemia-spiking mortality syndrome in broiler chickens with the use of homogenized brains containing arenaviruslike particles.

Severe hypoglycemia-spiking mortality syndrome was experimentally reproduced in broiler chicks. Inoculum was homogenized brains from 28-day-old commercial broiler chicks with central nervous system signs (50% [v/v] in phosphate-buffered saline with 2% fetal calf serum). Oral inoculations of 1.2 ml of the homogenate were given at 1 day of age to broiler chicks (n = 15). Fourteen days later, chicks were fasted and stressed with a 2-sec cool water spray. Six chicks (40%) developed clinical signs of spiking mortality syndrome and were severely hypoglycemic. Uninoculated control chicks (n = 15) from the same hatch, also fasted and stressed simultaneously, were unaffected. Examination of a banded fraction produced from the inoculum with the use of transmission electron microscopy with negative staining revealed viruslike particles indistinguishable from arenavirus particles stained and examined simultaneously. Avian encephalomyelitis virus was isolated by one of three laboratories attempting virus isolation with the use of embryonating chicken eggs.

Experimental reproduction of severe hypoglycemia and spiking mortality syndrome using field-derived and embryo-passaged preparations.

The clinical signs, enteritis, weight depression, and hypoglycemia of spiking mortality syndrome were experimentally reproduced in broiler breeders and broiler chicks. Inocula included 1) virus-like particles from intestines of chicks with spiking mortality syndrome that had been banded in a discontinuous Renograffin gradient, 2) homogenized darkling beetles collected from litter of farms where spiking mortality syndrome had occurred repeatedly, and 3) homogenized embryos which had been inoculated with the Renograffin-banded material. Arkansas variant infectious bronchitis virus and arenavirus-like particles were identified in the inocula. Serology on samples from surviving chicks suggested the presence of an avian encephalomyelitis virus in one of the inocula. One-day-old (n = 172) and 2.5-day-old (n = 30) chicks were inoculated orally, and some were also injected intraperitoneally or subcutaneously, with 0.5 ml of the inocula. Twelve to fourteen days postinoculation, chicks were fasted for 4-6 hours, then briefly stressed with a cool water spray. Within 1.5 hours, inoculated chicks began dying with severe hypoglycemia and clinical signs of spiking mortality syndrome. Body weights were significantly depressed. Uninoculated controls (n = 130) from the same hatches, also fasted and stressed, were unaffected clinically and were not hypoglycemic. One group (n = 52) of inoculated chicks exposed to a controlled lighting program was unaffected clinically, had significantly higher mean plasma glucose levels, and had significantly less body weight depression than chicks exposed to continuous lighting. We concluded that exposure to controlled amounts of light/darkness can ameliorate much of the hypoglycemia, mortality, and runting-stunting associated with spiking mortality syndrome of chickens. The significance of the viruses and virus-like particles detected in the inocula is currently under investigation.

Hypoglycemia, enteritis, and spiking mortality in Georgia broiler chickens: experimental reproduction in broiler breeder chicks.

The clinical signs, hypoglycemia, and mortality of "spiking mortality syndrome" were experimentally reproduced. Seven groups of day-old male primary broiler breeder chicks were orally inoculated with tissue and/or fecal-urate homogenates taken from field broilers with spiking mortality syndrome and from field broilers with enteritis and/or runting-stunting syndrome. All homogenates used as inocula were shown by transmission electron microscopy and negative staining to contain arenavirus-like particles. Inocula produced from field broilers with spiking mortality syndrome contained the highest numbers of the arena-virus-like particles and produced the highest percentage of hypoglycemic chicks 13-15 days postinoculation after a 5-to-9-hour fast. These homogenates also produced the most significant differences in mean plasma growth hormone and insulin-like growth factor-1 levels. The significance of the arenavirus-like particles is unknown but is currently being investigated.

PCR detection of group A bovine rotaviruses in feces.

A polymerase chain reaction (PCR) protocol has been developed for identification of bovine group A rotavirus infection in feces. Primers (20mers) complementary to 3' ends of double-stranded RNA genome segment 6 of bovine rotavirus NCDV strain were synthesized and used in PCR. Bovine rotavirus RNA from infected cell culture was employed to optimize the PCR protocol. Rotavirus-negative fecal samples were spiked with known quantities of bovine rotavirus, and the sensitivity of the PCR assay was determined. Fecal samples were extracted with phenol and treated to eliminate unidentified PCR inhibitor(s) in feces, and PCR was performed. PCR products were either visualized on ethidium bromide-stained agarose gels or detected by chemiluminescent hybridization. The sensitivity of the assay was 6 x 10(4) viral particles/ml of feces with ethidium bromide-stained agarose gel visualization or 6 x 10(2) viral particles/ml of feces with chemiluminescent hybridization. The PCR assay was applied to 18 fecal specimens from clinical cases. All 16 clinical samples that were positive for rotavirus by enzyme-linked immunosorbent assay (ELISA) or by ELISA and electron microscopy (EM) were positive by PCR. The 2 samples that were rotavirus negative by ELISA or by ELISA and EM were also negative on PCR analysis.

Detection of bluetongue virus in clinical samples by polymerase chain reaction.

A previously described bluetongue virus (BTV) serogroup polymerase chain reaction (PCR) assay was applied to clinical samples. The sensitivity of the BTV serogroup PCR was increased by the use of non-radioactive chemiluminescent hybridization. Unfractionated whole blood samples from rams experimentally inoculated with cell culture-adapted BTV-11 UC-8 were analyzed by virus isolation (VI) on Vero cells and PCR. VI and PCR were in agreement, with the exception of 3 blood samples that were VI negative and PCR positive. In semen spiked with BTV-11 UC-8, PCR detected as little as 1.6 x 10(2) plaque-forming units of BTV/ml of semen. BTV in the spleen of a sheep submitted for necropsy for suspect BTV infection was detected by both PCR and VI in embryonated chicken eggs. BTV PCR with nonradioactive chemiluminescent hybridization resulted in a level of sensitivity comparable to that of VI and likely more sensitive than VI on Vero cells for blood. This BTV PCR has great promise for rapid, sensitive, and specific detection of active BTV infection in a variety of clinical samples.