An Integrated Platform for in vivo Electrophysiology in Spatial Cognition Experiments.

Spatial cognition research requires behavioral paradigms that can distinguish between different navigational elements, such as allocentric (map-like) navigation and egocentric (e.g., body centered) navigation. To fill this need, we developed a flexible experimental platform that can be quickly modified without the need for significant changes to software and hardware. In this paper, we present this inexpensive and flexible behavioral platform paired with software which we are making freely available. Our behavioral platform serves as the foundation for a range of experiments, and though developed for assessing spatial cognition, it also has applications in the non-spatial domain of behavioral testing. There are two components of the software platform, 'Maze' and 'Stim Trigger'. Both programs can work in conjunction with electrophysiology acquisition systems, allowing for precise time stamping of neural events with behavior. The Maze program includes functionality for automatic reward delivery based on user defined zones. 'Stim Trigger' permits control of brain stimulation via any equipment that can be paired with an Arduino board. We seek to share our software and leverage the potential by expanding functionality in the future to meet the needs of a larger community of researchers.

Description of the ovarian microbiota of Aedes aegypti (L) Rockefeller strain.

Aedes aegypti is one of the vectors responsible for transmitting the viruses that cause dengue, Zika and chikungunya in the human population. Mosquitoes have bacterial communities in different organs, mainly in the midgut, but to a lesser extent in their reproductive organs, such as the ovaries, where replication and vertical transmission is decisive for dengue virus. These bacteria also influence metabolic and physiological processes such as ingestion and digestion of blood. In this study, aerobic bacterial communities associated with ovaries of A. aegypti Rockefeller strain were determined, describing their potential function during ovocitary development. The groups of mosquitoes were separated into three treatments: diet with 10% sugar solution, diet with blood supply, and blood feeding combined with tetracycline. The ovaries were extracted from the mosquitoes, and then put in enriched culture media (blood and nutritive agar) by direct inoculation, for subsequent isolation and macroscopic and microscopic characterization of the colonies. The taxonomic determination of bacterial isolates was achieved by sequence analysis of the 16S rRNA gene. A higher bacterial load was observed in the sugar feeding group (6 × 10³ CFU/ml) in contrast to the group fed only with blood, with and without an antibiotic (4.03-4.04 × 10³CFU/ml; 4.85-5.04 × 10³CFU/ml). As a result, a total of 35 colonies were isolated, of which 80% were gram-negative and 20% gram-positive; 72% were lactose negative and 8% lactose positive. Of the total bacteria, 83% had gamma hemolysis, 17% alpha hemolysis, and none presented beta hemolysis. After phenotypic and biochemical characterization, 17 isolates were selected for molecular identification. Only phyla Actinobacteria and Proteobacteria were found. Bacteria associated with ovaries of A. aegypti were mainly identified as belonging to the Serratia and Klebsiella genera. Some bacteria (Serratia marcescens, Pantoea dispersa and Klebsiella oxytoca) have wide biotechnological potential due to their entomopathogenic power and their bioactivity against different pathogens.

Afecciones respiratorias asociadas a factores ambientales y sanitarios en tres veredas de Guarne, Colombia, 2015 / Respiratory disease associated with environmental and health factors in three villages from Guarne, Colombia, 2015

RESUMEN Objetivo Determinar la asociación existente entre factores ambientales y sanitarios, y la prevalencia de afecciones respiratorias en los habitantes de las veredas El Sango, Romeral y Pastorcita del municipio de Guarne, Colombia, segundo semestre del año 2015. Materiales y Métodos El estudio fue de carácter descriptivo con análisis correlaciónales, en él se emplearon fuentes primarias de información, se diseñó una encuesta con preguntas cerradas. La población fue constituida por 493 habitantes, con edad ≥18 años. Se realizaron análisis descriptivos, bivariados con cálculos de prueba Chi 2 y Fischer, con RP y sus respectivos IC del 95%. Resultados Entre los más importantes, se encontró que el 6,5% de la población ha padecido en el último año, alguna afección respiratoria, las más frecuentes fueron: asma con 2,9% y EPOC con un 2,7%. Se encontraron asociaciones estadísticamente significativas entre padecer afecciones respiratorias y residir cerca de fábricas o industrias: RP 2,608 (IC 1,344-5,059) P<0,05; también se encontraron riesgos en personas que residen cerca de aguas negras o basureros: RP 2,333 (IC 1,278-4,257) P<0,05. Pudo encontrarse mayor prevalencia en personas cuyas casas tienen techos de barro, caña o esterilla: X2 3,214, P<0,05; así como vivir en la vereda Romeral: X2 7,032, P<0,05. Conclusiones La población de interés se encuentra expuesta a factores de contaminación ambiental (micro fábricas o industrias) y sanitaria (aguas negras y basureros) y que en efecto estos están asociados a la prevalencia de afecciones respiratorias en el sector.(AU)

ABSTRACT Objective To establish the association between environmental and health factors, as well as the prevalence of respiratory disease among the inhabitants of the villages of El Sango, Romeral and Pastorcita, located in the municipality of Guarne, Colombia, during the second half of 2015. Materials and Methods Descriptive correlational study. Primary sources of information were consulted and a survey with closed questions was designed. The population was made up of 493 inhabitants, aged ≥18 years. Descriptive bivariate analyzes were performed using Chi2 and Fischer test calculations, and including their corresponding PR and 95%CI. Results It was found that 6.5% of the population suffered from some respiratory condition within the past year, the most frequent being asthma (2.9%) and COPD (2.7%). Statistically significant associations were found between suffering from a respiratory disease and residing near factories or industries (PV: 2.608; 95% Cl: 1.344-5.059; p<0.05). People residing near sewage or garbage dumps were also at risk for respiratory disease (PR: 2.333; 95% CI: 1.278-4.257; p <0.05). A higher prevalence was observed in people whose houses had mud, cane or mat roofs (X2: 3.214; p <0.05) or lived in the village of Romeral (X2: 7.032; p <0.05). Conclusions The population of interest is exposed to environmental pollution (micro factories or industries) and sanitary factors (sewage and garbage), which are associated with the prevalence of respiratory diseases in the sector.(AU)

[Respiratory disease associated with environmental and health factors in three villages from Guarne, Colombia, 2015]. / Afecciones respiratorias asociadas a factores ambientales y sanitarios en tres veredas de Guarne, Colombia, 2015.

OBJECTIVE: To establish the association between environmental and health factors, as well as the prevalence of respiratory disease among the inhabitants of the villages of El Sango, Romeral and Pastorcita, located in the municipality of Guarne, Colombia, during the second half of 2015. MATERIALS AND METHODS: Descriptive correlational study. Primary sources of information were consulted and a survey with closed questions was designed. The population was made up of 493 inhabitants, aged ≥18 years. Descriptive bivariate analyzes were performed using Chi2 and Fischer test calculations, and including their corresponding PR and 95%CI. RESULTS: It was found that 6.5% of the population suffered from some respiratory condition within the past year, the most frequent being asthma (2.9%) and COPD (2.7%). Statistically significant associations were found between suffering from a respiratory disease and residing near factories or industries (PV: 2.608; 95% Cl: 1.344-5.059; p<0.05). People residing near sewage or garbage dumps were also at risk for respiratory disease (PR: 2.333; 95% CI: 1.278-4.257; p <0.05). A higher prevalence was observed in people whose houses had mud, cane or mat roofs (X2: 3.214; p <0.05) or lived in the village of Romeral (X2: 7.032; p <0.05). CONCLUSIONS: The population of interest is exposed to environmental pollution (micro factories or industries) and sanitary factors (sewage and garbage), which are associated with the prevalence of respiratory diseases in the sector.

OBJETIVO: Determinar la asociación existente entre factores ambientales y sanitarios, y la prevalencia de afecciones respiratorias en los habitantes de las veredas El Sango, Romeral y Pastorcita del municipio de Guarne, Colombia, segundo semestre del año 2015. MATERIALES Y MÉTODOS: El estudio fue de carácter descriptivo con análisis correlaciónales, en él se emplearon fuentes primarias de información, se diseñó una encuesta con preguntas cerradas. La población fue constituida por 493 habitantes, con edad ≥18 años. Se realizaron análisis descriptivos, bivariados con cálculos de prueba Chi 2 y Fischer, con RP y sus respectivos IC del 95%. RESULTADOS: Entre los más importantes, se encontró que el 6,5% de la población ha padecido en el último año, alguna afección respiratoria, las más frecuentes fueron: asma con 2,9% y EPOC con un 2,7%. Se encontraron asociaciones estadísticamente significativas entre padecer afecciones respiratorias y residir cerca de fábricas o industrias: RP 2,608 (IC 1,344-5,059) P<0,05; también se encontraron riesgos en personas que residen cerca de aguas negras o basureros: RP 2,333 (IC 1,278-4,257) P<0,05. Pudo encontrarse mayor prevalencia en personas cuyas casas tienen techos de barro, caña o esterilla: X2 3,214, P<0,05; así como vivir en la vereda Romeral: X2 7,032, P<0,05. CONCLUSIONES: La población de interés se encuentra expuesta a factores de contaminación ambiental (micro fábricas o industrias) y sanitaria (aguas negras y basureros) y que en efecto estos están asociados a la prevalencia de afecciones respiratorias en el sector.

Delivering kidney cancer care in rural Central and Southern Illinois: a telemedicine approach.

There is a growing body of experience and research suggesting that telemedicine (video conferencing, smart phones and online patient portals) could be the solution to addressing gaps in the provision of specialised healthcare in rural areas. The proposed role of telemedicine in providing needed services in hard to reach areas is not new. The United States Telecommunication Act of 1996 provided the initial traction for telemedicine by removing important economic and legal obstacles regarding the use of technology in healthcare delivery. This initial ruling has been supplemented by the availability of federal funding to support efforts aimed at developing telemedicine in underserved areas. In this paper, we explore one aspect of disease disparity pertinent to rural Illinois (kidney cancer incidence and mortality) and describe how we are planning to use an existing telemedicine program at Southern Illinois University School of Medicine (SIUSOM) to improve kidney cancer (Kca) care in rural Illinois. This represents an example of the possible role of telemedicine in addressing healthcare disparities in rural areas/communities and provides a description of general challenges and barriers to the implementation and maintenance of such systems.

Nitric oxide generation affects pro- and anti-angiogenic growth factor expression in primary human trophoblast.

OBJECTIVES: Preeclampsia is associated with reduced trophoblast placenta growth factor (PGF) expression, elevated soluble fms-like tyrosine kinase-1 (sFlt-1) and decreased bioactivity of nitric oxide (NO). Elevated sFlt-1 reduces bio-availability of PGF and vascular endothelial growth factor (VEGF) leading to maternal endothelial dysfunction. Although NO can regulate gene expression, its ability to regulate trophoblast expression of angiogenic growth factors is not known. STUDY DESIGN: Human primary term trophoblast and JEG-3 choriocarcinoma cells were cultured under 21%O(2) or 1%O(2) conditions in the presence or absence of NO donor (SNP) or inhibitor (L-NAME). Effects on PGF, VEGF and Flt-1 isoform mRNA expression were determined by quantitative real-time PCR. Changes in expression of soluble protein isoforms of FLT-1 was monitored by ELISA. RESULTS: Hypoxia decreased PGF mRNA but increased VEGF, sFlt-1 and Flt-1 mRNA expression in trophoblast. Generation of NO in trophoblast under 1%O(2) culture conditions significantly reversed sFlt-1 mRNA and protein expression, independent of mFlt-1. Conversely NO generation in hypoxic trophoblast increased VEGF and PGF mRNA expression. CONCLUSIONS: NO production in primary human trophoblast cultures had divergent effects on pro-angiogenic (PGF, VEGF) versus anti-angiogenic (sFlt-1) mRNA expression, resulting in an enhanced pro-angiogenic gene expression environment in vitro.

Neonatal corticosterone administration impairs adult eyeblink conditioning and decreases glucocorticoid receptor expression in the cerebellar interpositus nucleus.

Neonatal maternal separation alters adult learning and memory. Previously, we showed that neonatal separation impaired eyeblink conditioning in adult rats and increased glucocorticoid receptor (GR) expression in the cerebellar interpositus nucleus, a critical site of learning-related plasticity. Daily neonatal separation (1 h/day on postnatal days 2-14) increases neonatal plasma corticosterone levels. Therefore, effects of separation on GR expression in the interpositus and consequently adult eyeblink conditioning may be mediated by neonatal increases in corticosterone. As a first step in exploring a potential role for corticosterone in the neonatal separation effects we observed, we assessed whether systemic daily (postnatal days 2-14) corticosterone injections mimic neonatal separation effects on adult eyeblink conditioning and GR expression in the interpositus. Control uninjected animals were compared to animals receiving either daily corticosterone injections or daily injections of an equal volume of vehicle. Plasma corticosterone values were measured in a separate group of control, neonatally separated, vehicle injected, or corticosterone injected pups. In adulthood, rats underwent surgery for implantation of recording and stimulating electrodes. After recovery from surgery, rats underwent 10 daily sessions of eyeblink conditioning. Then, brains were processed for GR immunohistochemistry and GR expression in the interpositus nucleus was assessed. Vehicle and corticosterone injections both produced much larger increases in neonatal plasma corticosterone than did daily maternal separation, with the largest increases occurring in the corticosterone-injected group. Neonatal corticosterone injections impaired adult eyeblink conditioning and decreased GR expression in the interpositus nucleus, while the effects of vehicle injections were intermediate. Thus, while neonatal injections and maternal separation both produce adult impairments in learning and memory, these manipulations produce opposite changes in GR expression. This suggests an inverted U-shaped relationship may exist between both neonatal corticosterone levels and adult GR expression in the interpositus nucleus, and adult GR expression in the interpositus and eyeblink conditioning.

Chronic stress alters neural activity in medial prefrontal cortex during retrieval of extinction.

Chronic restraint stress produces morphological changes in medial prefrontal cortex and disrupts a prefrontally mediated behavior, retrieval of extinction. To assess potential physiological correlates of these alterations, we compared neural activity in infralimbic and prelimbic cortex of unstressed versus stressed rats during fear conditioning and extinction. After implantation of microwire bundles into infralimbic or prelimbic cortex, rats were either unstressed or stressed via placement in a plastic restrainer (3 h/day for 1 week). Rats then underwent fear conditioning and extinction while activity of neurons in infralimbic or prelimbic cortex was recorded. Percent freezing and neural activity were assessed during all phases of training. Chronic stress enhanced freezing during acquisition of conditioned fear, and altered both prelimbic and infralimbic activity during this phase. Stress did not alter initial extinction or conditioned stimulus (CS)-related activity during this phase. However, stress impaired retrieval of extinction assessed 24 h later, and this was accompanied by alterations in neuronal activity in both prelimbic and infralimbic cortex. In prelimbic cortex, unstressed rats showed decreased activity in response to CS presentation, whereas stressed rats showed no change. In infralimbic cortex, neurons in unstressed rats exhibited increased firing in response to the CS, whereas stressed rats showed no increase in infralimbic firing during the tone. Finally, CS-related firing in infralimbic but not prelimbic cortex was correlated with extinction retrieval. Thus, the stress-induced alteration of neuronal activity in infralimbic cortex may be responsible for the stress-induced deficit in retrieval of extinction.

Neonatal maternal separation alters the development of glucocorticoid receptor expression in the interpositus nucleus of the cerebellum.

Adverse early experience impairs adult learning and memory. Previously, we showed that neonatal maternal separation impaired eyeblink conditioning in adult male rats. This impairment was correlated with increases in glucocorticoid receptor expression in the posterior region of the cerebellar interpositus nucleus, a key structure in the neural circuitry controlling eyeblink conditioning. To begin to establish how separation results in altered glucocorticoid receptor expression in adulthood, we assessed the developmental pattern of glucocorticoid receptor expression in the interpositus nucleus in controls versus rats that had undergone maternal separation for 1h per day on postnatal days 2-14. Rat pups were exposed to either standard rearing (control) or maternal separation and glucocorticoid receptor expression was assessed at postnatal day 15, postnatal day 21, and adulthood. In control males, glucocorticoid receptor expression in the interpositus nucleus declined between postnatal days 15 and 21, then increased into adulthood. On postnatal day 15, there was less glucocorticoid receptor expression in the interpositus nucleus in males that were maternally separated than in controls. However, neonatal separation significantly attenuated the normal decline in the third postnatal week, resulting in significantly greater glucocorticoid receptor expression in the interpositus in separated males than in control rats at postnatal day 21. The developmental pattern of glucocorticoid receptor expression was not altered by maternal separation in female rats. Thus, maternal separation may impair learning and memory in adult males by altering normal developmental changes in glucocorticoid receptor expression.

Neonatal maternal separation-induced changes in glucocorticoid receptor expression in posterior interpositus interneurons but not projection neurons predict deficits in adult eyeblink conditioning.

Neonatal maternal separation impairs adult eyeblink conditioning. This impairment is correlated with increases in adult glucocorticoid receptor (GR) expression in the posterior interpositus nucleus [A.A. Wilber, C. Southwood, G. Sokoloff, J.E. Steinmetz, C.L. Wellman, Neonatal maternal separation alters adult eyeblink conditioning and glucocorticoid receptor expression in the interpositus nucleus of the cerebellum, Developmental Neurobiology 67 (2007) 1751-1764], a key structure in the neural circuitry controlling eyeblink conditioning. To further localize this effect, we assessed adult eyeblink conditioning and GR expression in projection versus interneurons in the interpositus of rats that had undergone standard rearing or maternal separation (1h/day) on postnatal days 2-14. At 3 months of age, interpositus neurons were labeled with the retrograde tracer biotinylated dextran amine (BDA). After delay eyeblink conditioning, brains were processed immunohistochemically for GR and BDA labeling of interpositus neurons. GR expression was quantified in BDA-labeled and unlabeled neurons. Neonatal maternal separation impaired adult eyeblink conditioning. Control rats had significantly less GR expression in posterior interpositus BDA-unlabeled versus BDA-labeled neurons, but this difference was absent in maternally separated rats. While neonatal separation significantly increased GR expression in BDA-labeled and unlabeled posterior interpositus neurons, only GR expression in non-BDA-labeled neurons was associated with eyeblink conditioning. Thus, neonatal maternal separation may alter interneuronal modulation of interpositus output neurons, producing deficits in adult eyeblink conditioning.

Dnase1l3 deficiency in lupus-prone MRL and NZB/W F1 mice.

Loss of deoxyribonuclease I (Dnase1) function is associated with systemic lupus erythematosus (SLE) in humans and mice; however, no coding mutations in Dnase1 are found in polygenic murine models. Instead, both MRL-lpr strains and NZB/W F1 hybrids are homozygous for T89I missense in the macrophage-DNASE, desoxyribonuclease I-like 3 (Dnase1l3). By in vitro expression studies, this substitution decreases this enzyme's nuclease activity against free DNA by only approximately twofold; however, the mutation has a greater effect on the capacity of media conditioned with Dnase1l3 to confer a barrier to liposomal gene transfection to HeLa cells. The 89I substitution decreases the Dnase1l3 barrier function in vitro by eightfold (P < 0.01). In splenocytes and BM-derived macrophages of SLE mice, while cellular Dnase1l3 levels are induced relative to C57BL/6 (control) mice, levels of FD-nuclease activity are similar. Finally, media conditioned by MRL and NZB/W F1 macrophages, relative to control, contains a weak interferon-gamma (IFN-gamma) inducible Dnase1l3-associated barrier to transfection. This barrier function is hypothesized to reflect the inability of SLE mice to degrade membrane-enveloped DNA-associated antigens, such as apoptotic bodies, which are predicted to stimulate the characteristic autoimmunity of SLE. Our results for these two generally independent models strongly suggest that Dnase1l3 deficiency increases the susceptibility of these mice to polygenic SLE.

Reverse microdialysis of a dopamine uptake inhibitor in the nucleus accumbens of alcohol-preferring rats: effects on dialysate dopamine levels and ethanol intake.

BACKGROUND: The mesolimbic dopamine (DA) system has been implicated in mediating the reinforcing actions of ethanol (EtOH). This study examines the effects of local perfusion of the DA uptake inhibitor GBR12909 (GBR) on (1) DA levels in the nucleus accumbens (NAc) and (2) EtOH drinking in alcohol-preferring rats. METHODS: Stable drinking of a 15% (v/v) EtOH solution (minimum of 0.75 g/kg body weight) was established in daily 1 hr limited access sessions. Rats were then implanted with bilateral guide cannulae aimed 4 mm above the NAc. After recovery from surgery, concentric microdialysis probes (2 mm dialysis membrane surface) were inserted into the NAc. Most placements were in the shell or overlapping both shell and core. Two days later, the probes were perfused at 1.0 microl/min with artificial cerebral spinal fluid (aCSF) for at least a 90 min washout period followed by collection of five basal samples over 150 min. Rats were then perfused with either aCSF alone or 10, 25, 100, or 200 microM of GBR for 240 min on the first day of microdialysis. During the last 60 min of the drug treatment phase, rats were given their scheduled access to 15% EtOH. All rats were then perfused with aCSF for the last 90 min of the experiment. The following day, the procedure was repeated, but animals that received aCSF on the first day were given a dose of GBR and rats given GBR on the first day received only aCSF. RESULTS: GBR perfusion increased extracellular NAc DA levels dose dependently to more than 800% of basal levels at 100 to 200 microM but failed to alter EtOH intake (p > 0.05, paired t test) at any concentration tested. Moreover, after 100 microM of GBR perfusion had terminated, the extracellular levels of DA in the NAc remained elevated for approximately 24 hr (790% of day 1 basal; p < 0.05). The increase in dialysate DA levels observed during GBR perfusion with 100 microM was significantly greater for EtOH-experienced rats than for EtOH-naïve rats [F(7,59) = 14.85, p < 0.0001, analysis of variance, Student-Newman-Keuls post hoc test]. CONCLUSIONS: The results suggest (1) that EtOH drinking experience induces neuroadaptations that increase DA release in the NAc, and (2) that additional elevation in synaptic levels of DA in the NAc does not influence the maintenance of ongoing alcohol drinking under scheduled access conditions in alcohol-preferring animals.

Studies on some trace and minor elements in blood. A survey of the Kalpakkam (India) population. Part III: Studies on dietary intake and its correlation to blood levels.

In our studies on elemental levels in blood of the Kalpakkam population, it was found that the reference values for many elements were normal, but some deficiency with respect to Se was noticed. As a followup study, the dietary ingredients of the local population were analyzed for trace and minor elements to assess the dietary intake of these elements. Details of the analytical methods developed using the technique of inductively coupled plasma-mass spectrometry (ICP-MS) and atomic absorption spectrometry (AAS) have been described. The dietary intake of many of these trace and minor elements were found to be quite adequate according to the recommended dietary allowance (RDA) levels prescribed, except for Se and Zn. The dietary intake of Se was found to be in the range 20-50 micrograms/d (as opposed to the RDA of 50-200 micrograms/d), whereas the intake of Zn was found to be in the range 8-10 mg/d (as opposed to the RDA of 15 mg/d). Although the deficiency of Se intake was reflected in the blood, that of Zn was not, probably owing to the high level of homeostasis for this element. Fish and egg were found to be rich sources of Se, followed by cereals and pulses, which were found to be the major sources of Zn.