Fasting Modulates GABAergic Synaptic Transmission to Arcuate Kisspeptin Neurons in Female Mice.

It is well-established that the hypothalamic-pituitary-gonadal (HPG) axis is suppressed due to negative energy balance. However, less information is available on whether kisspeptin neuronal activity contributes to fasting-induced responses. In the present study, female and male mice were fasted for 24 hours or provided food ad libitum (fed group) to determine whether acute fasting is sufficient to modulate kisspeptin neuronal activity. In female mice, fasting attenuated luteinizing hormone (LH) and prolactin (PRL) serum levels and increased follicle-stimulating hormone levels compared with the fed group. In contrast, fasting did not affect gonadotropin or PRL secretion in male mice. By measuring genes related to LH pulse generation in micropunches obtained from the arcuate nucleus of the hypothalamus (ARH), we observed that fasting reduced Kiss1 mRNA levels in female and male mice. In contrast, Pdyn expression was upregulated only in fasted female mice, whereas no changes in the Tac2 mRNA levels were observed in both sexes. Interestingly, the frequency and amplitude of the GABAergic postsynaptic currents recorded from ARH kisspeptin neurons (ARHKisspeptin) were reduced in 24-hour fasted female mice but not in males. Additionally, neuropeptide Y induced a hyperpolarization in the resting membrane potential of ARHKisspeptin neurons of fed female mice but not in males. Thus, the response of ARHKisspeptin neurons to fasting is sexually dependent with a female bias, associated with changes in gonadotropins and PRL secretion. Our findings suggest that GABAergic transmission to ARHKisspeptin neurons modulates the activity of the HPG axis during situations of negative energy balance.

Hypothalamic Kisspeptin Neurons as a Target for Whole-Cell Patch-Clamp Recordings.

Kisspeptins are essential for the maturation of the hypothalamic-pituitary-gonadal (HPG) axis and fertility. Hypothalamic kisspeptin neurons located in the anteroventral periventricular nucleus and rostral periventricular nucleus, as well as the arcuate nucleus of the hypothalamus, project to gonadotrophin-releasing hormone (GnRH) neurons, among other cells. Previous studies have demonstrated that kisspeptin signaling occurs through the Kiss1 receptor (Kiss1r), ultimately exciting GnRH neuron activity. In humans and experimental animal models, kisspeptins are sufficient for inducing GnRH secretion and, consequently, luteinizing hormone (LH) and follicle stimulant hormone (FSH) release. Since kisspeptins play an essential role in reproductive functions, researchers are working to assess how the intrinsic activity of hypothalamic kisspeptin neurons contributes to reproduction-related actions and identify the primary neurotransmitters/neuromodulators capable of changing these properties. The whole-cell patch-clamp technique has become a valuable tool for investigating kisspeptin neuron activity in rodent cells. This experimental technique allows researchers to record and measure spontaneous excitatory and inhibitory ionic currents, resting membrane potential, action potential firing, and other electrophysiological properties of cell membranes. In the present study, crucial aspects of the whole-cell patch-clamp technique, known as electrophysiological measurements that define hypothalamic kisspeptin neurons, and a discussion of relevant issues about the technique, are reviewed.

Gestational exposure to continuous light impairs the development of the female reproductive system in adult Wistar rat offspring.

INTRODUCTION: It has been suggested that maternal exposure to constant light during the gestational period could be considered as a chronic stressor, impairing offspring development by interfering in neuroendocrine and behavior responses. OBJECTIVE: This study aimed to evaluate whether maternal exposure to continuous light during pregnancy affects the adult reproductive system in the female offspring. MATERIALS AND METHODS: Pregnant Wistar rats were allocated into light-dark (LD) group, exposed to light and dark photoperiod during gestation, and the light-light (LL) group, exposed to a photoperiod of constant light during gestation. After birth, pups were maintained under normal light-dark photoperiod until adulthood. At postnatal day 90, blood was collected from the female offspring, to analyze plasma luteinizing hormone (LH) and progesterone levels, and the uterus and ovaries were harvested for morphometric, histological, and oxidative stress evaluations. RESULTS AND DISCUSSION: Female exposure to continuous light during the intrauterine period resulted in the adult reduction of LH and increased progesterone plasma levels, and uterine injuries a higher number of endometrial glands and reduced levels of antioxidant enzymes, such as glutathione reductase and glutathione S-transferase. In these experimental conditions, gestational continuous light exposure disturbs sex hormone balance and reduces the antioxidant enzymatic activity in the uterus of female offspring in adult life.

Pattern of gonadotropin secretion along the estrous cycle of C57BL/6 female mice.

The pattern of gonadotropin secretion along the estrous cycle was elegantly described in rats. Less information exists about the pattern of gonadotropin secretion in gonad-intact mice, particularly regarding the follicle-stimulating hormone (FSH). Using serial blood collections from the tail-tip of gonad-intact C57BL/6 mice on the first day of cornification (transition from diestrus to estrus; hereafter called proestrus), we observed that the luteinizing hormone (LH) and FSH surge cannot be consistently detected since only one out of eight females (12%) showed increased LH levels. In contrast, a high percentage of mice (15 out of 21 animals; 71%) exhibited LH and FSH surges on the proestrus when a single serum sample was collected. Mice that exhibited LH and FSH surges on the proestrus showed c-Fos expression in gonadotropin-releasing hormone- (GnRH; 83.4% of co-localization) and kisspeptin-expressing neurons (42.3% of co-localization) of the anteroventral periventricular nucleus (AVPV). Noteworthy, mice perfused on proestrus, but that failed to exhibit LH surge, showed a smaller, but significant expression of c-Fos in GnRH (32.7%) and AVPVKisspeptin (14.0%) neurons. Finally, 96 serial blood samples were collected hourly in eight regular cycling C57BL/6 females to describe the pattern of LH and FSH secretion along the estrous cycle. Small elevations in LH and FSH levels were detected at the time expected for the LH surge. In summary, the present study improves our understanding of the pattern of gonadotropin secretion and the activation of central components of the hypothalamic-pituitary-gonadal axis along the estrous cycle of C57BL/6 female mice.

Neurobehavioral assessment of Danio reriointoxicated by Mercury and the use of Mercurius solubilis

Mercury is used in various industrial. Part of Mercury's industrial waste is discharged into the environment, rivers and their tributaries, thus contaminating aquatic animals. Aim:to evaluate Mercury-induced behavioral changes in Zebrafish (Danio rerio) by the analysis of locomotor activity and parameters related to neurotoxicity and to verify whether ultra-diluted substances can decrease neurobehavioral effects and toxic. Methodology:The fishes were separated into 4 monitoring aquariums with 8 fishes each, with temperature, pH controlled, until the time of the toxicological experiments. 0.5 mL of Mercury 6cH, 30cH and distilled water (positive control) were added per liter of water in each aquarium containing 6 liters of water, then 3 mL of medication per aquarium, the white control received no medication and the toxic agent. After 1 hour the drugs were added, toxic mercury (200 µg/L), 4 mL per aquarium was added and remained so for 24 hours. All the experiment was run in blind, and the drugs identified by codes. The animals were subjected to behavioral tests (Open Field-locomotion; Vertical Open Field for neurotoxicity evaluation and Light and Dark Test), and each stage was recorded for later evaluation of movements and neurobehavioral changes. ANOVA was performed, followed by Tukey test, with p <0.05. Results: Mercury produced an anxiogenic effect in animals that were submitted to it without medication. In the vertical open field, there was an increase in erratic movements (1.25 ± 1.0) and tremors (0.87 ± 0.35) compared to the control (0.12 ± 0.35 and 0.25 ± 0.46 respectively), proving the toxic effect. Fishes which received the medication at 6 cH and 30 ch showed tremors and erratic movements similar to control. Conclusion:200 µg/L mercury in water can cause neurobehavioral disturbances in fishes, and animals receiving Mercurius6 cH and 30 cH ultra-diluted drug did not show neurotoxicity.

Can CBCT change the level of confidence of oral maxillofacial surgeons in mandibular third molar management?

Abstract This study analyzed the impact of CBCT on the level of confidence in diagnostic and treatment thinking in mandibular lower molar (M3M) clinical management. Thirty cases for which panoramic radiographs and CBTC images were available were selected and classified according to radiologic signs indicating the proximity of the M3M to the mandibular canal (interruption of the radiopaque borders of the canal of the mandibular canal wall, darkening of the roots, and diversion or narrowing of the canal, n = 10 for each classification). Twelve oral and maxillofacial surgeons (OMS) contributed to this study by answering two questionnaires. The first questionnaire contained a clinical description of the case and a panoramic radiograph. After 30 days, a second questionnaire with the same clinical illustrations and tomographic multiplanar reconstruction images was administered. Both questionnaires asked specialists to rate diagnostic confidence, the surgical complexity, chosen treatment, and surgical confidence. In approximately 40% of answers, CBCT images had a positive impact on ratings of diagnostic confidence and treatment thinking confidence, and in 24.4%, they increased the surgical complexity score. There was no change in the treatment plan following the use of CBCT, but the CBCT examination was a determining factor for diagnosis and treatment planning in 72.8% of the answers CBCT improved the confidence level in diagnostic and treatment thinking of the M3M management while also increasing the perceived level of surgical complexity. The findings of this study support the need to consider using CBCT in diagnosis and treatment planning for M3Ms with radiographic signs such as darkening of the roots, interruption of the radiopaque borders of the mandibular canal, or deviation of the mandibular canal and narrowing of the roots.

Distinct effects of growth hormone deficiency and disruption of hypothalamic kisspeptin system on reproduction of male mice.

Growth hormone (GH) deficiency is a common cause of late sexual maturation and fertility issues. To determine whether GH-induced effects on reproduction are associated with alterations in hypothalamic kisspeptin system, we studied the male reproduction in two distinct GH deficiency mouse models. In the first model, mice present GH deficiency secondary to arcuate nucleus of the hypothalamus (ARH) lesions induced by posnatal monosodium glutamate (MSG) injections. MSG-induced ARH lesions led to significant reductions in hypothalamic Ghrh mRNA expression and consequently growth. Hypothalamic Kiss1 mRNA expression and Kiss1-expressing cells in the ARH were disrupted in the MSG-treated mice. In contrast, kisspeptin immunoreactivity remained preserved in the anteroventral periventricular and rostral periventricular nuclei (AVPV/PeN) of MSG-treated mice. Importantly, ARH lesions caused late sexual maturation and infertility in male mice. In our second mouse model, we studied animals profound GH deficiency due to a loss-of-function mutation in the Ghrhr gene (Ghrhrlit/lit mice). Interestingly, although Ghrhrlit/lit mice exhibited late puberty onset, hypothalamic Kiss1 mRNA expression and hypothalamic kisspeptin fiber density were normal in Ghrhrlit/lit mice. Despite presenting dwarfism, the majority of Ghrhrlit/lit male mice were fertile. These findings suggest that spontaneous GH deficiency during development does not compromise the kisspeptin system. Furthermore, ARH Kiss1-expressing neurons are required for fertility, while AVPV/PeN kisspeptin expression is sufficient to allow maturation of the hypothalamic-pituitary-gonadal axis in male mice.

Neonatal metformin short exposure inhibits male reproductive dysfunction caused by a high-fat diet in adult rats.

Metformin (Met) is widely used to control blood glucose levels and acts on various organs, including reproductive tissues, to improve reproductive and lifespan. This study evaluated whether neonatal Met exposure prevented male reproductive dysfunction caused by being overweight during adulthood. Randomized Wistar rat pups received an intraperitoneal injection from postnatal days (PNDs) 1 to 12of saline (Sal; 0.9% NaCl/day in 2mL/kg) or Met (100 mg/kg/day in 2 mL/kg). From PNDs 60 to 90, the animals received a regular (R; 4.5% fat; Sal R and Met R groups) or a high-fat (HF; 35% fat; Sal HF and Met HF groups) diet. At PND 90, all animals were euthanized to evaluate their biometric and reproductive parameters. The Sal and Met groups with R showed similar body weights, however, the HF diet increased the body weight in both groups. The Sal HF group showed testicular damage regarding in antioxidant status and inflammatory profile in the epididymal cauda. The HF diet reduced Leydig and Sertoli cells numbers, with lower sperm quality. The Met R animals showed positive reproductive programming, due to improved antioxidant defense, inflammatory biomarkers, and sperm morphology. Met HF prevented HF diet damage to reproductive organs and sperm morphology, but not to sperm motility. Early Met exposure positively affected the male reproductive system of adult rats, preventing reproductive HF disorders. STATEMENT OF NOVELTY AND SIGNIFICANCE: Metformin is used to control type 2 diabetes mellitus and can act to improve metabolism and lifespan. Metformin avoidance is recommended during pregnancy, but there is no information regarding its use when breastfeeding. For the first time, we showed in this current study that metformin positively acts in the male reproductive tissues and helps involved in later life. These data showed a better antioxidant defense and anti-inflammatory profile of Metformin animals than Saline animals and might directly improve reproductive organs morphophysiology and sperm morphology. Also, the neonatal Met application programs the male reproduction to counterbalance damages from an obesogenic environment in later life.

Pulmonary Emphysema Impairs Male Reproductive Physiology Due To Testosterone and Oxidative Stress Imbalance in Mesocricetus auratus.

This study evaluated whether pulmonary emphysema affects sperm quality, male reproductive organs, and testosterone levels in adult male hamsters. Mesocricetus auratus males (130-150 g) were subdivided into a control group (C group) and an emphysema group (E group). The C group received an intratracheal instillation of saline solution (0.3 mL/100 g of body weight), and the E group received papain (40 mg/100 g of body weight). After 60 days, the biometric, pulmonary, and reproductive parameters of each group were evaluated. The E group developed pulmonary emphysema, which decreased body weight and sperm quality compared to the C group. In oxidative stress-related assays, lipid peroxidation was increased in the testis and epididymis (caput and cauda) in the E group compared with the C group. However, only the caput epididymis showed a reduction in glutathione levels. Pulmonary emphysema also affected the testicle by inducing an increase in abnormal seminiferous tubules, accompanied by a decrease in seminiferous epithelium height. Spermatogenesis kinetics were also modified by pulmonary emphysema. The number of Leydig and Sertoli cells decreased in the E group, accompanied by an increase in the nuclear volume of Leydig cells. Testosterone concentration was increased in the E group. Similarly, pulmonary emphysema altered epididymal components in all regions. In conclusion, pulmonary emphysema affected the reproductive system in this experimental model, as shown by testicular and epididymal morphophysiology changes, hormonal alteration, and oxidative stress imbalance, inducing the loss of correct function in the male reproductive system.

Diagnostic efficacy of different cone beam computed tomography scanning protocols in the detection of chemically simulated external root resorption.

OBJECTIVES: The aim of this study was to evaluate the efficacy of different high-resolution cone beam computed tomography protocols in the diagnosis of chemically simulated external root resorption (ERR). STUDY DESIGN: ERR was simulated in 3 sizes by using an acid decalcification protocol in the cervical, middle, and apical thirds of 30 single-rooted premolars. Four scans of each tooth were acquired with different voxel sizes: 0.080, 0.085, 0.120, and 0.133 mm. The images were analyzed by 2 blinded evaluators for the presence of ERR. The evaluation of diagnostic performance was based on receiver operating characteristic (ROC) curve analysis. RESULTS: No statistically significant differences were found in the detection of ERR with regard to lesion sizes or the different root thirds. Areas under the ROC curve were 0.901, 0.892, 0.887, and 0.767 for voxel sizes of 0.080, 0.085, 0.120, and 0.133 mm, respectively. Voxel sizes of 0.080 and 0.085 mm presented significantly larger areas under the ROC curve compared with the voxel size of 0.133 mm. There were no statistically significant differences among the protocols with the use of voxel sizes of 0.080, 0.085, and 0.120 mm. CONCLUSIONS: Cone beam computed tomography examinations with higher-resolution protocols demonstrated better performance in the diagnosis of ERR. Further investigations using simulations that more closely resemble the actual ERR process are warranted.

Physical and physiological quality of seeds of Campo Grande Stylosanthes coated with different binder materials / Qualidade física e fisiológica de sementes de Stylosanthes Campo Grande revestidas com diferentes materiais cimentantes

ABSTRACT: The binder materials used in seed coating provide adherence of the filler materials onto the seeds. Adequate concentrations of the binder promote good physical characteristics of coating while not impairing seed physiological characteristics. Thus, the objective of this study was to evaluate the type and the concentration of binder materials that allow the best physical and physiological characteristics in coated seeds of Campo Grande Stylosanthes. Seed coating was carried out in a coating pan, using sand as the filling material. The treatments consisted of 2 types of binders mixed in 3 ratios (PVA glue and gum arabic glue, diluted in water, 3:1, 2:1, and 1:1), and uncoated seeds as control. The coated seeds were evaluated for physical and physiological characteristics. The laboratory and greenhouse experiments were arranged in a completely randomized design. Results showed that at the different concentrations used this study, neither the PVA glue nor the gum arabic glue had any effect on the physiological quality of Campo Grande Stylosanthes seeds. However, PVA glue in the ratio of 2:1 provided the greatest surface area, maximum diameter, minimum diameter, and sphericity to coated seeds.

RESUMO: No processo de revestimento, os materiais cimentantes são os responsáveis por conferir a adesão dos materiais de enchimento às sementes. Concentrações adequadas de material cimentante promovem revestimento com boas características físicas e que não prejudiquem as características fisiológicas das sementes. Assim, o objetivo desse trabalho foi verificar o tipo e a concentração de material cimentante que possibilite as melhores características físicas e fisiológicas em sementes revestidas de estilosantes Campo Grande. O revestimento foi realizado em drageadora, utilizando areia como material de enchimento. Os tratamentos foram compostos por 2 tipos de materiais cimentantes em 3 proporções (cola cascorez extra e cola goma arábica, diluídas em água, 3:1, 2:1 e 1:1), além das sementes não revestidas, como controle. Após o revestimento, as sementes foram avaliadas quanto às características físicas e fisiológicas. O delineamento adotado foi inteiramente casualizado em laboratório e em casa de vegetação. Verificou-se que as colas cascorez extra e goma arábica não interferiram na qualidade fisiológica das sementes de estilosantes Campo Grande nas diferentes concentrações utilizadas. Entretanto, a cola cascorez extra na proporção 2:1 proporcionou maior área, diâmetro máximo, diâmetro mínimo e esfericidade nas sementes revestidas.

Effect of packaging materials and storage environment on fennel seedling growth / Efeito da embalagem e do ambiente de armazenamento no crescimento de plântulas de erva doce

The objective of the present study was to assess the influence of the packaging material (cotton cloth bag, multifolium paper and glass container) and storage environment (laboratory and cold chamber conditions) on fennel seedling growth. Seeds were placed in different packaging materials and exposed to the two environments for twelve months. Assessments were made before storage and at every two months and the following were determined: total emergence, emergence speed index, seedling fresh and dry matter. A completely randomized experimental design was used, with four replications in a 2 x 3 x 6+1 factorial design in split split plots, where the plot was the storage environment (laboratory and cold chamber conditions); the split plot was the storage (cotton cloth bag, multifolium paper and glass container) and the split plot was the storage periods (2, 4, 6, 8, 10 and 12 months) and plus an additional treatment (before storage). The results showed that the fennel seedlings had superior growth when their seeds were placed in the glass packaging material, regardless of the storage environment.

O objetivo deste trabalho foi avaliar a influência da embalagem (saco de algodão, papel multifoliado e vidro) e do ambiente de armazenamento (condições de laboratório e câmara fria) no crescimento de plântulas de erva doce. Para isso, as sementes foram acondicionadas nas diferentes embalagens e expostas aos dois ambientes durante doze meses. As avaliações foram feitas antes do armazenamento e a cada dois meses, tendo sido determinado: emergência total, índice de velocidade de emergência, massa fresca e seca das plântulas. Utilizou-se o delineamento experimental inteiramente casualizado, com quatro repetições, num esquema fatorial 2 x 3 x 6 + 1 em parcelas subsubdivididas, no qual a parcela foi o ambiente de armazenamento (condições de laboratório e câmara fria); a subparcela as embalagens (saco de algodão, papel multifoliado e vidro) e a subsubparcela os períodos de armazenamento (2, 4, 6, 8, 10 e 12 meses) e mais um tratamento adicional (antes do armazenamento). Pelos resultados, demonstrou-se que as plântulas de erva doce apresentaram crescimento superior quando suas sementes foram acondicionadas na embalagem de vidro, independente do ambiente de armazenamento.

Genome-wide DNA methylation profile in the peripheral blood of cocaine and crack dependents

Objective: Cocaine use disorders (CUDs) represent a major public health problem in many countries. To better understand the interaction between the environmental modulations and phenotype, the aim of the present study was to investigate the DNA methylation pattern of CUD patients, who had concomitant cocaine and crack dependence, and healthy controls. Methods: We studied DNA methylation profiles in the peripheral blood of 23 CUD patients and 24 healthy control subjects using the Illumina Infinium HumanMethylation450 BeadChip arrays. Results: Comparison between CUD patients and controls revealed 186 differentially methylated positions (DMPs; adjusted p-value [adjP] < 10-5) related to 152 genes, with a subset of CpGs confirmed by pyrosequencing. DNA methylation patterns discriminated CUD patients and control groups. A gene network approach showed that the EHMT1, EHMT2, MAPK1, MAPK3, MAP2K1, and HDAC5 genes, which are involved in transcription and chromatin regulation cellular signaling pathways, were also associated with cocaine dependence. Conclusion: The investigation of DNA methylation patterns may contribute to a better understanding of the biological mechanisms involved in CUD.

Soil patterns and foliar standards for two cocoa clones in the States of Espírito Santo and Bahia, Brazil / Padrões de solo e normas foliares para dois clones de cacau cultivados nos Estados do Espírito Santo e Bahia, Brasil

ABSTRACT: The objective of this research was to establish soil patterns and foliar standards for clonal crops of cocoa CCN51 and PS1319 in the Atlantic regions within the northern Espírito Santo and southern Bahia States. Samples were collected from September to October 2015, in 45 high productive plots, being 23 plots of clone CCN51 and 22 of clone PS1319. Sufficiency ranges for soil and foliar standards were recommended for cocoa clones CCN51 and PS1319 planted in the Northern Espírito Santo and Southern Bahia states. Differences in the contents of foliar macronutrients K, Ca, Mg, S and micronutrients Fe and Mn, were verified between clones CCN51 and PS1319. Differences reported for standards from different geographical regions confirmed that these standards need to be regional and specific for different cocoa clones.

RESUMO: O objetivo deste trabalho foi estabelecer padrões de solo e padrões foliares para culturas clonais de cacau CCN51 e PS1319 nas regiões do Atlântico no norte do Espírito Santo e no sul da Bahia. As amostras foram coletadas de setembro a outubro de 2015, em 45 parcelas de alta produtividade, sendo 23 parcelas do clone CCN51 e 22 do clone PS1319. As faixas de suficiência para solo e padrões foliares foram sugeridas para os clones de cacau CCN51 e PS1319 plantados no norte do Espírito Santo e no sul da Bahia. Diferenças nos teores dos macronutrientes foliares K, Ca, Mg, S e micronutrientes Fe e Mn foram verificadas entre os clones CCN51 e PS1319. As diferenças relatadas para padrões de diferentes regiões geográficas permitem confirmar que esses padrões precisam ser regionais e específicos para diferentes clones de cacau.

Bisphenol A Exposure Impairs Epididymal Development during the Peripubertal Period of Rats: Inflammatory Profile and Tissue Changes.

Bisphenol A (BPA) is a synthetic non-steroidal oestrogen used in the production of plastics. BPA can cause alterations in the endocrine system of human beings and animals at varied stages of development. During puberty, altered morphological, sexual behaviour and completion of the epididymal development occur. Therefore, this study aimed to evaluate the effects of BPA on epididymal development during the peripubertal period of rats. Male Wistar rats were treated with BPA via gavage at doses of 20 µg/kg or 200 µg/kg per day [post-natal day (PND] 36-66). The control group received the vehicles under the same conditions. Feed and water were provided ad libitum. On PND 67, the epididymis was removed, weighed, divided into caput/corpus and cauda sections. It was then used for sperm count determination; histopathological and stereological evaluation; inflammatory cell enzymatic profiling (myeloperoxidase activity - MPO; N-acetylglucosaminidase - NAG); immunohistochemistry for IL-6; and evaluation of superoxide anion levels and malondialdehyde (MDA). Exposure to BPA at 200 µg/kg caused a significant increase of MPO activity and immunoreactivity to IL-6 (interleukin-6) as well as remodelling of tissue components in the caput/corpus and cauda regions of the epididymis. Under these experimental conditions, it is concluded that BPA alters post-natal epididymal development.

Co-expression network of neural-differentiation genes shows specific pattern in schizophrenia.

BACKGROUND: Schizophrenia is a neurodevelopmental disorder with genetic and environmental factors contributing to its pathogenesis, although the mechanism is unknown due to the difficulties in accessing diseased tissue during human neurodevelopment. The aim of this study was to find neuronal differentiation genes disrupted in schizophrenia and to evaluate those genes in post-mortem brain tissues from schizophrenia cases and controls. METHODS: We analyzed differentially expressed genes (DEG), copy number variation (CNV) and differential methylation in human induced pluripotent stem cells (hiPSC) derived from fibroblasts from one control and one schizophrenia patient and further differentiated into neuron (NPC). Expression of the DEG were analyzed with microarrays of post-mortem brain tissue (frontal cortex) cohort of 29 schizophrenia cases and 30 controls. A Weighted Gene Co-expression Network Analysis (WGCNA) using the DEG was used to detect clusters of co-expressed genes that were non-conserved between adult cases and controls brain samples. RESULTS: We identified methylation alterations potentially involved with neuronal differentiation in schizophrenia, which displayed an over-representation of genes related to chromatin remodeling complex (adjP = 0.04). We found 228 DEG associated with neuronal differentiation. These genes were involved with metabolic processes, signal transduction, nervous system development, regulation of neurogenesis and neuronal differentiation. Between adult brain samples from cases and controls there were 233 DEG, with only four genes overlapping with the 228 DEG, probably because we compared single cell to tissue bulks and more importantly, the cells were at different stages of development. The comparison of the co-expressed network of the 228 genes in adult brain samples between cases and controls revealed a less conserved module enriched for genes associated with oxidative stress and negative regulation of cell differentiation. CONCLUSION: This study supports the relevance of using cellular approaches to dissect molecular aspects of neurogenesis with impact in the schizophrenic brain. We showed that, although generated by different approaches, both sets of DEG associated to schizophrenia were involved with neocortical development. The results add to the hypothesis that critical metabolic changes may be occurring during early neurodevelopment influencing faulty development of the brain and potentially contributing to further vulnerability to the illness.

Hiperparatireoidismo como causa de demência reversível em idosos / Hyperparathyroidism as a cause of reversible dementia in elderly

Introdução: demência é uma síndrome caracterizada pelo declínio progressivo e global das funções cognitivas. Existem diversas condições reversíveis que podem causá-la ou mimetizá-la. Objetivos: investigar o hiperparatireoidismo (HPT) como causa de demência e seu potencial de reversibilidade. Métodos: avaliação clínico-laboratorial de 60 idosos com suspeita de demência encaminhados ao Ambulatório Affonso Silviano Brandão da Faculdade de Ciências Médicas de Minas Gerais e HospitalUniversitário São José ? Belo Horizonte. Os níveis séricos do cálcio total, 25 hidroxivitamina D e paratormônio (PTH), no diagnóstico, foram, respectivamente, de 12,4 e 13,2 mg/dL; 14 e 10 ng/mL; e 227 e 302 pg/mL. Resultados: a idade média foi de 73 ± 8 anos; 70% femininos; sendo duas pacientes com níveis elevados de cálcio e PTH e baixos de vitamina D. Apresentavam também litíase renal, alterações da função cognitiva,sintomas depressivos, osteoporose com fraturas e nódulo em região cervical visível ao ultrassom e hipercaptantes à Sestamibi-99mTc por cintilografia. A causa provável do HPT, epidemiologicamente, era um adenoma. Confirmado o diagnóstico, optou-se pelo tratamento cirúrgico pela técnica minimamente invasiva, com monitorização do PTH intraoperatório (PTH?IO). Foram submetidos pré e pós-operatório (seis meses), ao MEEM e CAMDEX-R. Obtiveram melhora dos escores, no primeiro, de 11 em ambas as pacientes para 16 na paciente A e 15 na paciente B; e no segundo, inicialmente de 65 na paciente A e de 63 na paciente B para 82 em ambas as pacientes. Conclusões: o HPTdeve ser considerado na avaliação dos quadros de demência.

Introduction: dementia is a syndrome characterized by a progressive decline in cognitive and global functions. There are several reversible conditions that may cause it or mimic it. Objectives: to investigate the hyperparathyroidism (HPT) as the cause of dementia andtheir potential reversibility. Methods: clinical and laboratory evaluation of 60 elderly people with suspected dementia referred to the Affonso Silviano Brandão Outpatient Ambulatory at the Medical Sciences College of Minas Gerais and São José University Hospital - Belo Horizonte. The total serum calcium, 25-hydroxyvitamin D, and parathyroid hormone (PTH) in the diagnosis were 12.4, and 13.2 mg/dL; 14 and 10 ng/mL; and 227 and 302 pg/ml, respectively. Results: The mean age was 73 ± 8 years; 70% female; including two patients with high levels of calcium and PTH and low levels of vitamin D. They also presented low kidney stones, changes in cognitive function, depressive symptoms, osteoporosis with fractures,and visible lump in cervical region ultrasound, and hypercaptant to Sestamibi-99mTc by scintigraphy. The probable cause of HPT, epidemiologically, was an adenoma. Once the diagnosis was confirmed, we opted for surgical treatment through a minimally invasivetechnique, with intraoperative PTH monitoring (PTH-IO). These patients underwent pre and postoperative (six months) to the MMSE and CAMDEX-R. They showed improved scores from 11 points in the first inboth patients to 16 in patient A and 15 in patient B; and inthe second, from 65 in patient A and 63 in patient B to 82in both patients. Conclusions: HPT should be consideredin the evaluation of dementia cases.

Genome-wide DNA methylation profile of leukocytes from melanoma patients with and without CDKN2A mutations.

Melanoma is a highly aggressive cancer, accounting for up to 75% of skin cancer deaths. A small proportion of melanoma cases can be ascribed to the presence of highly penetrant germline mutations, and approximately 40% of hereditary melanoma cases are caused by CDKN2A mutations. The current study sought to investigate whether the presence of germline CDKN2A mutations or the occurrence of cutaneous melanoma would result in constitutive genome-wide DNA methylation changes. The leukocyte methylomes of two groups of melanoma patients (those with germline CDKN2A mutations and those without CDKN2A mutations) were analyzed together with the profile of a control group of individuals. A pattern of DNA hypomethylation was detected in the CDKN2A-negative patients relative to both CDKN2A-mutated patients and controls. Additionally, we delineated a panel of 90 CpG sites that were differentially methylated in CDKN2A-mutated patients relative to controls. Although we identified a possible constitutive epigenetic signature in CDKN2A-mutated patients, the occurrence of reported SNPs at the detected CpG sites complicated the data interpretation. Thus, further studies are required to elucidate the impact of these findings on melanoma predisposition and their possible effect on the penetrance of CDKN2A mutations.