It Is All in the Right Amygdala: Increased Synaptic Plasticity and Perineuronal Nets in Male, But Not Female, Juvenile Rat Pups after Exposure to Early-Life Stress.

Early-life stress (ELS) is associated with increased vulnerability to mental disorders. The basolateral amygdala (BLA) plays a critical role in fear conditioning and is extremely sensitive to ELS. Using a naturalistic rodent model of ELS, the limited bedding paradigm (LB) between postnatal days 1-10, we previously documented that LB male, but not female preweaning rat pups display increased BLA neuron spine density paralleled with enhanced evoked synaptic responses and altered BLA functional connectivity. Since ELS effects are often sexually dimorphic and amygdala processes exhibit hemispheric asymmetry, we investigated changes in synaptic plasticity and neuronal excitability of BLA neurons in vitro in the left and right amygdala of postnatal days 22-28 male and female offspring from normal bedding or LB mothers. We report that LB conditions enhanced synaptic plasticity in the right, but not the left BLA of males exclusively. LB males also showed increased perineuronal net density, particularly around parvalbumin (PV) cells, and impaired fear-induced activity of PV interneurons only in the right BLA. Action potentials fired from right BLA neurons of LB females displayed slower maximal depolarization rates and decreased amplitudes compared with normal bedding females, concomitant with reduced NMDAR GluN1 subunit expression in the right BLA. In LB males, reduced GluA2 expression in the right BLA might contribute to the enhanced LTP. These findings suggest that LB differentially programs synaptic plasticity and PV/perineuronal net development in the left and right BLA. Furthermore, our study demonstrates that the effects of ELS exposure on BLA synaptic function are sexually dimorphic and possibly recruiting different mechanisms.SIGNIFICANCE STATEMENT Early-life stress (ELS) induces long-lasting consequences on stress responses and emotional regulation in humans, increasing vulnerability to the development of psychopathologies. The effects of ELS in a number of brain regions, including the amygdala, are often sexually dimorphic, and have been reproduced using the rodent limited bedding paradigm of early adversity. The present study examines sex differences in synaptic plasticity and cellular activation occurring in the developing left and right amygdala after limited bedding exposure, a phenomenon that could shape long-term emotional behavioral outcomes. Studying how ELS selectively produces effects in one amygdala hemisphere during a critical period of brain development could guide further investigation into sex-dependent mechanisms and allow for more targeted and improved treatment of stress-and emotionality-related disorders.

Experience of Adversity during a First Lactation Modifies Prefrontal Cortex Morphology in Primiparous Female Rats: Lack of Long Term Effects on a Subsequent Lactation.

Reproductive experience is associated with morphological and functional plasticity in brain areas important for cognitive and emotional responses, including the infralimbic (IL) medial prefrontal cortex (mPFC). Here we examined whether suboptimal conditions during a first lactation could modify lactation-induced morphological IL mPFC changes, leading to alterations in stress responses and attention and whether any observed effects would persist into a second lactation. Reduced availability of bedding and nesting material (LB) was used to induce unfavorable conditions in primiparous (P) mothers. In normal bedding (NB) conditions, P mothers exhibited high spine number and density on postpartum day (PPD)10, which greatly decreased 2 weeks after weaning of their pups. In contrast, P-LB mothers had a lower spine number and density on PPD10, which markedly increased after weaning. LB exposure did not modify stress responsiveness to a ferret odor on PPD5 in primiparous or in multiparous (M) females. Number of errors and trials to criterion in the attention set shifting task were not modified by a history of adversity in multiparous females, although this group tended to exhibit higher attentional abilities than M-NB females. These results suggest that adversity acutely reduces morphological plasticity in the maternal mPFC during lactation, an effect that is not associated with significant changes in stress responses and/or glucocorticoid production. Medial PFC morphological changes induced by LB subside during a subsequent lactation as does the effect of maternity itself.

Increased Hypothalamic Projections to the Lateral Hypothalamus and Responses to Leptin in Rat Neonates From High Fat Fed Mothers.

The lateral hypothalamus (LHA) is a central hub in the regulation of food intake and metabolism, as it integrates homeostatic and hedonic circuits. During early development, maturing input to and output from the LHA might be particularly sensitive to environmental dietary changes. We examined the effects of a maternal high fat diet (HFD, 60% Kcal in fat) on the density of hypothalamic projections to the orexin (ORX-A) field of the LHA in 10 day-old (PND10) rat pups using retrograde labeling with fluorescent microspheres. We also compared responsiveness of phenotypically identified LHA neurons to leptin administration (3 mg/kg, bw) between pups from control (CD) or high fat (HFD) fed mothers on PND10 and 15-16, at the onset of independent feeding. HFD pups exhibited a higher density of LHA projections (p = 0.05) from the ventromedial hypothalamus (VMH) compared to CD pups and these originated from both SF-1 and BDNF-positive neurons in the VMH. Increased circulating leptin levels in HFD pups, particularly on PND15-16 was consistent with enhanced pSTAT3 responses to leptin in the orexin (ORX-A) field of the LHA, with some of the activated neurons expressing a GABA, but not CART phenotype. ORX-A neurons colocalizing with pERK were significantly higher in PND15-16 HFD pups compared to CD pups, and leptin-induced increase in pERK signaling was only observed in CD pups. There was no significant effect of leptin on pERK in HFD pups. These results suggest that perinatal maternal high fat feeding increases hypothalamic projections to the ORX-A field of the LHA, increases basal activation of ORX-A neurons and direct responsiveness of LHA neurons to leptin. Since these various LHA neuronal populations project quite heavily to Dopamine (DA) neurons in the ventral tegmental area, they might participate in the early dietary programming of mesocorticolimbic reward circuits and food intake.

Gating of the neuroendocrine stress responses by stressor salience in early lactating female rats is independent of infralimbic cortex activation and plasticity.

In early lactation (EL), stressor salience modulates neuroendocrine stress responses, but it is unclear whether this persists throughout lactation and which neural structures are implicated. We hypothesized that this process is specific to EL and that the infralimbic (IL) medial prefrontal cortex (mPFC) might provide a critical link between assessment of threat and activation of the hypothalamo-pituitary-adrenal (HPA) axis in EL. We measured neuroendocrine responses and neuronal Fos induction to a salient (predator odor) or non-salient (tail pinch) psychogenic stressor in EL and late lactation (LL) females. We found that EL females exhibited a large response to predator stress only in the presence of pups, while responses to tail pinch were reduced independently of pup presence. In LL, HPA axis responses were independent of pup presence for both stressors and only responses to tail pinch were modestly reduced compared to virgins. Intracerebral injection of the local anesthetic bupivacaine (BUP) (0.75%; 0.5 µl/side) in the IL mPFC did not differentially affect neuroendocrine responses to predator odor in virgin and EL females, suggesting that lactation-induced changes in this structure might not regulate stressor salience for the HPA axis. However, the IL mPFC displayed morphological changes in lactation, with significant increases in dendritic spine numbers and density in EL compared to LL and virgin females. EL females also showed improved performance in the attention set-shifting task (AST), which could reflect early plasticity in the IL mPFC at a time when rapid adaptation of the maternal brain is necessary for pup survival.

Measurement of cortisol in saliva: a comparison of measurement error within and between international academic-research laboratories.

OBJECTIVE: Hundreds of scientific publications are produced annually that involve the measurement of cortisol in saliva. Intra- and inter-laboratory variation in salivary cortisol results has the potential to contribute to cross-study inconsistencies in findings, and the perception that salivary cortisol results are unreliable. This study rigorously estimates sources of measurement variability in the assay of salivary cortisol within and between established international academic-based laboratories that specialize in saliva analyses. One hundred young adults (Mean age: 23.10 years; 62 females) donated 2 mL of whole saliva by passive drool. Each sample was split into multiple- 100 µL aliquots and immediately frozen. One aliquot of each of the 100 participants' saliva was transported to academic laboratories (N = 9) in the United States, Canada, UK, and Germany and assayed for cortisol by the same commercially available immunoassay. RESULTS: 1.76% of the variance in salivary cortisol levels was attributable to differences between duplicate assays of the same sample within laboratories, 7.93% of the variance was associated with differences between laboratories, and 90.31% to differences between samples. In established-qualified laboratories, measurement error of salivary cortisol is minimal, and inter-laboratory differences in measurement are unlikely to have a major influence on the determined values.

Inhibition of anandamide hydrolysis dampens the neuroendocrine response to stress in neonatal rats subjected to suboptimal rearing conditions.

Exposure to stress during early development can exert profound effects on the maturation of the neuroendocrine stress axis. The endocannabinoid (ECB) system has recently surfaced as a fundamental component of the neuroendocrine stress response; however, the effect of early-life stress on neonatal ECB signaling and the capacity to which ECB enhancement may modulate neonatal stress responses is relatively unknown. The present study assessed whether exposure to early-life stress in the form of limited access to nesting/bedding material (LB) from postnatal (PND) day 2 to 9 alters neuroendocrine activity and hypothalamic ECB content in neonatal rats challenged with a novel immobilization stressor. Furthermore, we examined whether inhibition of fatty acid amide hydrolase (FAAH), the enzyme responsible for the degradation of anandamide (AEA) affects neuroendocrine responses in PND10 pups as a function of rearing conditions. Neonatal rats showed a robust increase in corticosterone (CORT) and adrenocorticotropin hormone (ACTH) secretion in response to immobilization stress, which was significantly blunted in pups reared in LB conditions. Accordingly, LB pups exhibited reduced stress-induced Fos immunoreactivity in the paraventricular nucleus of the hypothalamus, with no significant differences in hypothalamic ECB content. Administration of the FAAH inhibitor URB597 (0.3 mg/kg, ip) 90 min prior to immobilization stress significantly dampened stress-induced CORT release, but only in pups reared in LB conditions. These results suggest that rearing in restricted bedding conditions dampens the neuroendocrine response to stress, while augmenting AEA mitigates stress-induced alterations in glucocorticoid secretion preferentially in pups subjected to early-life stress.

Distribución genotipo-específico del virus papiloma humano entre hombres y mujeres de Caracas, Venezuela / Genotype-specific distribution of HPV between men and women od Caracas, Venezuela

Determinar la distribución de VPH entre hombres y mujeres con técnicas de biología molecular con la finalidad de aportar información epidemiológica para la evaluación de la utilidad del uso de las vacunas contra VPH en la población. Se procesaron muestras de hisopados endocervicales, uretrales, balano prepuciales, biopsias y cepillados cervicales, para extracción de ADN: Kit AxyPrep Body Fluid Viral DNA/RNA Miniprep (Axygen), para la amplificación: Kit INNO-LIPA HPV Genotyping Extra. Se recolectaron datos en el sistema electrónico INFOLAB. 1 545 pacientes en total fueron separados por sexo, grupo etario y amplificación de ADN-VPH. 77,6 por ciento de los pacientes del sexo masculino y 67,9 por ciento del sexo femenino resultaron positivos para la amplificación de ADN-VPH. Se encontró un pico de positividad entre los 30 a 34 años de edad (24,3 por ciento) al igual que para el sexo masculino (20,0 por ciento). En el sexo femenino 50,6 por ciento,0 por ciento a otros genotipos. En el sexo masculino 41,3 por ciento corresponde a VPH-AR, 34,6 por ciento VPH-BR y un 24,2 por ciento a otros genotipos. Los genotipos AR detectados con más frecuencia fueron VPH-52/51/16 y los de BR VPH-6/11. El cáncer cervical es un problema de salud pública. El tamizaje con citología no ha tenido impacto sobre la prevalencia, se sugiere el uso de pruebas más sensibles. Se debe evaluar el uso de la vacuna en conjunto con otras estrategias, realizando un análisis identificando las debilidades y fortalezas, con la finalidad de establecer programas de prevención óptimos

Determine the distribution of HPV in men and women with Molecular Biology techniques in order to provide epidemiological information to evaluate the usefulness of the HPV vaccine in the population. Samples of endocervical swabs, urethral, preputial, biopsies and cervical scrapes for DNA extraction: Body Fluid Kit AxyPrep Viral DNA / RNA Miniprep (Axygen) for the amplification: INNO-LIPA HPV Kit Extra Genotyping. Data were collected in the electronic system Infolab. 1 545 patients in total were separated by sex, age group and HPV DNA amplification. 77.6 percent of male patients and 67.9 percent of females were positive for HPV DNA amplification. We found a positive peak between 30 and 34 years of age (24.3 percent) as well as for males (20.0 percent). Among females 50.6 percent of genotypes identified corresponded to HR-HPV, 22.4 percent to HPV-LR, 27.0 percent for other genotypes. In males, 41.3 percent were HR-HPV, HPV-LR 34.6 percent and 24.2 percent other genotypes. AR genotypes detected more frequently were HPV- 52/51/16 and LR were HPV-6/11. Cervical cancer is a public health problem. Cytology screening has had no impact on the prevalence, we suggest the use of more sensitive tests. It is necessary to evaluate the use of the vaccine in conjunction with other strategies, analyzing the strengths and weaknesses identified, in order to establish optimal prevention programs