Role of the hypothalamus in ghrelin effects on reproduction: sperm function and sexual behavior in male mice.

In brief: Ghrelin signals to the hypothalamus inhibit reproduction during times of food scarcity. In this study, we demonstrate that ghrelin impairs sperm quality in male mice. Abstract: Ghrelin (GHRL) is an orexigenic peptide that has been investigated as one of the signals responsible for the reproductive performance of mammals under fluctuating metabolic conditions. Central GHRL administration impairs spermatogenesis in mice by regulating the hypothalamic-pituitary-gonadal axis function. In the present study, the hypothalamus role as a mediator of GHRL effects on sperm fertilizing capacity and male sexual behavior was evaluated. After 42 days of hypothalamic GHRL infusion or artificial cerebrospinal fluid, in vitro and in vivo sperm fertilizing capacity, testicular α-tubulin, speriolin gene expression and spermatic α-tubulin protein were evaluated. Hypothalamic expression of genes Kiss1, Gpr54 and Gnrh was also studied. The second group of animals was infused with one time only GHRL or artificial cerebrospinal fluid into the hypothalamus to evaluate the effects on sexual behavior. Results demonstrated that chronic GHRL administration to male mice significantly increased the percentages of pre-implantation embryo loss and the number of post-implantation embryo loss. In relation to the gene expression, our results show a relative decrease of Kiss1, Gpr54 and Spatc1. Although no significant differences were observed in the quantitative expression of α-tubulin protein, qualitative changes in its expression pattern were observed. In addition, a dual effect on sexual behavior was observed: 40% of the treated animals showed a significant reduction in the number of mounts and intromissions, while a 60% showed a significant decrease in ejaculation latency vs control animals. In conclusion, our results provide evidence that central GHRL administration possibly induces failure in embryo development and/or implantation in the females mated with treated males, possibly because of a negative effect in the α-tubulin pattern.

Levels of energy supplementation for heifers in Tifton 85 pasture on carcass characteristics, internal organs and meat quality.

The objective of this study was to evaluate the effect of different levels of concentrate supplementation on carcass characteristics of heifers finished on Tifton 85 pasture. Thirty-two cross-breed heifers (Charolais and Nellore) on average aged 25.51 months and weighing 298 kg were used in a completely randomized experiment. The animals were distributed uniformly to receive four different levels of concentrates based on the body weight: 0.0; 0.4; 0.8 and 1.2%. Animals were slaughtered at 29 months of age. There was increasing linear effect of levels energy supplementation on: slaughter weight, hot and cold carcass weights,hot and cold dressing percentage, thigh thickness, carcass compactness, and L* and b* color values. It was observed quadratic effect of levels of energy supplementation on the percentage of fat in the carcass. Different levels of energy supplementation did not modify the accumulation of vital organs and visceral fat or the sensory attributes of the meat. Higher levels of energy supplementation promote increases in carcass traits of heifers when finished on Tifton 85 pasture.

Inter- and trans-generational effects of gestational ghrelin imbalance on development and reproduction in the mouse.

CONTEXT AND AIMS: We have demonstrated that ghrelin (Ghrl) participates in fetal programming, since intragestational hyperghrelinaemia increased pup's growth and a Ghrl-receptor antagonist accelerated offspring's sexual maturation and impaired their adult reproductive function. Now, we aim to analyse if these phenotypic changes (found in F1) also occurred in F2 and/or F3 generations. METHODS: We treated mice dams (F0), with 4nmol/animal/day of Ghrl or 6nmol/animal/day of an antagonist [Ant:(d -Lys3)GHRP6] from day 1 of pregnancy until delivery. When F1 female pups reached adulthood, they were paired to obtain F2, and subsequently, F2 females were paired to obtain F3. Parameters evaluated in F2 and F3 pups were: growth, physical development, neurobiological maturation, puberty onset and in adulthood, reproductive function. KEY RESULTS: The F2 and F3 Ant groups showed a significant increase in litter size. Although no differences were detected in the weight of these pups at birth, in adulthood, they were heavier. At F3, pups from the Ant group showed advanced incisors eruption and eye opening compared to controls. Furthermore, F3 male pups from the Ant group showed earlier testis descent, although in adulthood, these males exhibited reduced sperm concentration in comparison to Ghrl. No differences were detected in F2 or F3 females regarding puberty onset or reproduction. CONCLUSIONS AND IMPLICATIONS: Some fetal programming effects of Ghrl seen in F1, also appeared transgenerationally. Since many women at reproductive age suffer from conditions with reduced Ghrl levels (i.e. obesity or polycystic ovarian syndrome), these results could be relevant to the health of their descendants.

Therapeutic hypothermia for the treatment of neonatal hypoxia-ischemia: sex-dependent modulation of reactive astrogliosis.

Therapeutic hypothermia (TH) is the standard treatment for neonatal hypoxia-ischemia (HI) with a time window limited up to 6 h post injury. However, influence of sexual dimorphism in the therapeutic window for TH has not yet been elucidated in animal models of HI. Therefore, the aim of this study was to investigate the most effective time window to start TH in male and female rats submitted to neonatal HI. Wistar rats (P7) were divided into the following groups: NAÏVE and SHAM (control groups), HI (submitted to HI) and TH (submitted to HI and TH; 32ºC for 5 h). TH was started at 2 h (TH-2 h group), 4 h (TH-4 h group), or 6 h (TH-6 h group) after HI. At P14, animals were subjected to behavioural tests, volume of lesion and reactive astrogliosis assessments. Male and female rats from the TH-2 h group showed reduction in the latency of behavioral tests, and decrease in volume of lesion and intensity of GFAP immunofluorescence. TH-2 h females also showed reduction of degenerative cells and morphological changes in astrocytes. Interestingly, females from the TH-6 h group showed an increase in volume of lesion and in number of degenerative hippocampal cells, associated with worse behavioral performance. Together, these results indicate that TH neuroprotection is time- and sex-dependent. Moreover, TH started later (6 h) can worsen volume of brain lesion in females. These data indicate the need to develop specific therapeutic protocols for each sex and reinforce the importance of early onset of the hypothermic treatment.

Is seminal quality worsening? A 20-year experience in Córdoba, Argentina.

PURPOSE: To assess the possible variations in semen quality during the last 20 years in Córdoba, Argentina, and to identify possible causal lifestyle or genitourinary factors. METHODS: Retrospective study of 23,130 patients attending an andrology laboratory. The 20-year period (2001-2020) was divided into four quinquenniums. Seminal parameters (sperm concentration, motility, morphology, viability, and membrane functional integrity) were classified as normal or abnormal according to WHO, and results were expressed as percentage of patients abnormal for each parameter per quinquennium. In addition, the percentage of patients per quinquennium exposed to the different risk factors (daily alcohol and/or tobacco consumption; occupational exposure to heat or toxics; history of parotitis or varicocele; and high body mass index, BMI) was reported. RESULTS: Patients included in our study did not show impairment in seminal quality over time. Beyond a transient decrease in normozoospermia in the second and third quinquennium, possibly explained by a parallel increase in teratozoospermia, other important parameters of the spermogram did not change. In fact, abnormalities in sperm concentration (oligozoospermia), total sperm count, viability and response to hypoosmotic test showed a decreasing trend over time. On the other hand, parotitis, varicocele, morbid obesity and regular exposition to heat/toxics were the factors more frequently associated with semen abnormalities; the last two increased their frequency over the study period. CONCLUSION: The population included in this study did not show a clear impairment in semen quality during the last 20 years. The decreasing patterns found were associated with high BMI and exposure to heat/toxics.

Do aging, drinking, and having unhealthy weight have a synergistic impact on semen quality?

PURPOSE: To evaluate if age, alcohol consumption, and body mass index (BMI) have synergistic effects on seminal quality, and to rank these factors based on their impact on semen. METHODS: Retrospective study of 9464 patients attending an andrology laboratory. Data on patients' age and daily alcohol intake were provided by the patients. BMI was recorded in the laboratory. Seminal parameters evaluated were volume, sperm concentration and total count, motility, morphology, viability, nuclear maturity, and membrane functional integrity. RESULTS: All the seminal parameters evaluated were affected by the synergistic interaction Age x BMI, suggesting that this combination is more potent in affecting semen quality. The variables sperm morphology and nuclear maturity seemed to be especially susceptible since they were affected by the three synergistic interactions. In the logistic regression analysis, age was the most powerful factor since it impacted first on five of the nine parameters, impacting mainly on sperm motility, viability, and morphology, with no effects on sperm count. On the contrary, BMI impacted first in sperm concentration and total sperm count; which was confirmed also by the logistic predictions analysis. Alcohol consumption impacted first on membrane functional integrity and nuclear maturity. A J-shaped association between BMI or alcohol consumption with semen quality was found in the multivariate analysis. CONCLUSION: The factors considered in this study showed a synergistic negative impact on semen quality, being age and unhealthy weight the most important ones. Reducing the exposure to lifestyle risk factors may be promising for improving sperm quality in infertile patients.

Recategorisation of body mass index to achieve andrological predictive power: a study in more than 20 000 patients.

The aim of this study was to recategorise body mass index (BMI) in order to classify patients according to their risk of semen abnormalities. Patients (n=20563) presenting at an andrology laboratory were classified into five groups according to BMI: underweight (BMI <20kg m-2), normal weight (BMI 20-24.9kg m-2), overweight (BMI 25-29.9kg m-2), obese (BMI 30-39.9kg m-2) and morbidly obese (BMI >40kg m-2). Semen quality was evaluated to determine: (1) differences between groups using analysis of variance (ANOVA); (2) the chances of semen abnormalities (using generalised linear models, Chi-squared tests and odds ratios); (3) reference BMI values with andrological predictive power (multivariate conglomerate analyses and multivariate analysis of variance (MANOVA)); and (4) expected values of abnormalities for each new group resulting from BMI recategorisation. Morbidly obese and underweight patients exhibited the highest decrease in semen quality and had higher chances of semen abnormalities. The smallest number of sperm abnormalities was found at a BMI of 27kg m-2. Four reference values were identified, recategorising BMI into four groups according to their risk of semen abnormalities (from lowest to highest risk): Group1,BMI between 20 and 32kg m-2; Group2, BMI <20 and BMI >32-37kg m-2; Group3, BMI >37-42kg m-2; and Group4, BMI >42kg m-2. A BMI <20 or >32kg m-2 is negatively associated with semen quality; these negative associations on semen quality increase from a BMI >37kg m-2 and increase even further for BMI >42kg m-2. The BMI recategorisation in this study has andrological predictive power.

Emergence profile customization technique during implant transfer.

This case report aimed to describe the clinical considerations in oral rehabilitation with prostheses on implants in the anterior region, using emergence profile customization technique during implant transfer. A patient presented with a missing left upper central incisor and with not satisfactory esthetic in the other upper incisors. After diagnosis, a treatment plan was elaborated: implant placement in this region; ceramic laminates for teeth 11 and 22, and full crown for tooth 12. During the impression procedure, the implant transfer was customized by copying the gingival profile. The prostheses were made using IPS e.max ceramic system and luted with resin cement. The patient was submitted to annual follow-up examinations that did not show any irregularity or deficiency in the prostheses. The technique performed is a viable alternative that can be used in oral rehabilitation with implants involving anterior teeth because it allows a predictable level of adaptation.

Reproductive performance of male mice after hypothalamic ghrelin administration.

It has been demonstrated that food intake and reproductive physiology are both simultaneously modulated to optimize reproductive success under fluctuating metabolic conditions. Ghrelin (GHRL) is an orexigenic peptide identified as the endogenous ligand of the growth hormone secretagogue receptor that is being investigated for its potential role on reproduction. Considering that data available so far are still limited and characterization of GHRL action mechanism on the reproductive system has not been fully elucidated, we studied the participation of hypothalamus in GHRL effects on sperm functional activity, plasma levels of gonadotropins and histological morphology in mice testes after hypothalamic infusion of 0.3 or 3.0 nmol/day GHRL or artificial cerebrospinal fluid (ACSF) at different treatment periods. We found that GHRL 3.0 nmol/day administration for 42 days significantly reduced sperm concentration (GHRL 3.0 nmol/day = 14.05 ± 2.44 × 106/mL vs ACSF = 20.33 ± 1.35 × 106/mL, P < 0.05) and motility (GHRL 3.0 nmol/day = 59.40 ± 4.20% vs ACSF = 75.80 ± 1.40%, P < 0.05). In addition, histological studies showed a significant decrease percentage of spermatogonia (GHRL 3.0 nmol/day = 6.76 ± 0.68% vs ACSF = 9.56 ± 0.41%, P < 0.05) and sperm (GHRL 3.0 nmol/day = 24.24 ± 1.92% vs ACSF = 31.20 ± 3.06%, P < 0.05). These results were associated with a significant reduction in luteinizing hormone and testosterone plasma levels (P < 0.05). As GHRL is an orexigenic peptide, body weight and food intake were measured. Results showed that GHRL increases both parameters; however, the effect did not last beyond the first week of treatment. Results presented in this work confirm that central GHRL administration impairs spermatogenesis and suggest that this effect is mediated by inhibition of hypothalamic-pituitary-gonadal axis.

Influence of visible light on cutaneous hyperchromias: Clinical efficacy of broad-spectrum sunscreens.

INTRODUCTION: Cutaneous hyperchromias are disorders of skin pigmentation involving increased melanin production and its irregular accumulation in skin cells. The use of sunscreens is fundamental for the control of hyperchromias by reducing the stimulation of pigmentation, as melanin synthesis is mainly stimulated by solar radiation. Many studies have demonstrated that visible light can induce significant skin damage. Considering the effects of visible light, effective photoprotection should not be limited only to UV protection but should also involve visible and infrared protection. OBJECTIVE: The aim of this study was to evaluate the efficacy of UV-VIS sunscreens in protecting skin against damages caused by solar radiation and the influence of visible light on the appearance of cutaneous hyperchromias. METHODS: Forty volunteers aged 18 to 39 years with skin hyperpigmentation participated in the study. To evaluate the efficacy of the formulations developed, the percentage of hyperpigmented area was evaluated using high-resolution images-Visioface® Quick (Courage-Khazaka, Germany) and the analysis of epidermal pigmentation was performed by RCM-Vivascope® 1500 (Lucid, USA). Also, the melanin index was determined using the Mexameter® M X16 colorimeter (Courage-Khazaka, Germany). RESULTS: The developed formulations were effective in the reduction in melanin index, epidermal pigmentation, and percentage of hyperpigmented area. CONCLUSION: Finally, this study discusses how the combination of UV filters and pigments can protect the skin from solar radiation and reduces skin hyperpigmentations.

Stress distribution in delayed replanted teeth splinted with different orthodontic wires: a three-dimensional finite element analysis.

AIM: The aim was to evaluate the biomechanical behavior of the supporting bony structures of replanted teeth and the periodontal ligament (PDL) of adjacent teeth when orthodontic wires with different mechanical properties are applied, with three-dimensional finite element analysis. MATERIALS AND METHODS: Based on tomographic and microtomographic data, a three-dimensional model of the anterior maxilla with the corresponding teeth (tooth 13-tooth 23) was generated to simulate avulsion and replantation of the tooth 21. The teeth were splinted with orthodontic wire (Ø 0.8 mm) and composite resin. The elastic modulus of the three orthodontic wires used, that is, steel wire (FA), titanium-molybdenum wire (FTM), and nitinol wire (FN) were 200 GPa, 84 GPa, and 52 GPa, respectively. An oblique load (100 N) was applied at an angle of 45° on the incisal edge of the replanted tooth and was analyzed using Ansys Workbench software. The maximum (σmax) and minimum (σmin) principal stresses generated in the PDL, cortical and alveolar bones, and the modified von Mises (σvM) values for the orthodontic wires were obtained. RESULTS: With regard to the cortical bone and PDL, the highest σmin and σmax values for FTM, FN, and FA were checked. With regard to the alveolar bone, σmax and σmin values were highest for FA, followed by FTM and FN. The σvM values of the orthodontic wires followed the order of rigidity of the alloys, that is, FA > FTM > FN. CONCLUSION: The biomechanical behavior of the analyzed structures with regard to all the three patterns of flexibility was similar.

Role of ghrelin in fertilization, early embryo development, and implantation periods.

In order to clarify the physiological role of ghrelin in gestation, we evaluated the effects of administration of exogenous ghrelin (2 or 4 nmol/animal per day) or its antagonist (6 nmol/animal per day of (d-Lys3)GHRP6) on fertilization, early embryo development, and implantation periods in mice. Three experiments were performed, treating female mice with ghrelin or its antagonist: i) starting from 1 week before copulation to 12 h after copulation, mice were killed at day 18 of gestation; ii) since ovulation induction until 80 h later, when we retrieved the embryos from oviducts/uterus, and iii) starting from days 3 to 7 of gestation (peri-implantation), mice were killed at day 18. In experiments 1 and 3, the antagonist and/or the highest dose of ghrelin significantly increased the percentage of atrophied fetuses and that of females exhibiting this finding or a higher amount of corpora lutea compared with fetuses (nCL/nF) (experiment 3: higher nCL/nF-atrophied fetuses: ghrelin 4, 71.4-71.4% and antagonist, 75.0-62.5% vs ghrelin 2, 46.2-15.4% and control, 10-0.0%; n=7-13 females/group; P<0.01). In experiment 2, the antagonist diminished the fertilization rate, and both, ghrelin and the antagonist, delayed embryo development (blastocysts: ghrelin 2, 62.5%; ghrelin 4, 50.6%; and antagonist, 61.0% vs control 78.4%; n=82-102 embryos/treatment; P<0.0001). In experiment 3, additionally, ghrelin (4 nmol/day) and the antagonist significantly diminished the weight gain of fetuses and dams during pregnancy. Our results indicate that not only hyperghrelinemia but also the inhibition of the endogenous ghrelin effects exerts negative effects on the fertilization, implantation, and embryo/fetal development periods, supporting the hypothesis that ghrelin (in 'adequate' concentrations) has a physiological role in early gestational events.

Acrobatic training prevents motor deficits and neuronal loss in male and female rats following chronic cerebral hypoperfusion.

BACKGROUND: Chronic cerebral hypoperfusion in vascular dementia leads to memory and motor deficits; Physical exercise improves these aspects and promotes neuroprotection. Sexual dimorphism may significantly influence both ischemic and exercise outcomes. AIMS: The aim of this study was to investigate the effects of 2VO (Two-Vessel occlusion) and the acrobatic training on motor function, functional performance, and tissue loss in male and female rats. METHODS: Male and female rats were randomly divided into 4 groups: sham acrobatic, sham sedentary, 2VO acrobatic and 2VO sedentary. After 45 days of 2VO surgery, the animals received 4 weeks of acrobatic training. At the end, open field, beam balance and horizontal ladder tests were performed. Brain samples were taken for histological and morphological evaluation. RESULTS: Spontaneous motor activity in the open field was not affected by 2VO, on the other hand, an impairment in forelimb placement was observed after 2VO and acrobatic training prevented errors and improved hindlimb placement. Neuronal loss was found in the motor cortex and striatum after 2VO, especially in females, which was prevented by acrobatic training. CONCLUSION: Mild motor damage was found in animals after 2VO when refined movement was evaluated, probably associated to neuronal death in the motor cortex and striatum. The acrobatic exercise showed a neuroprotective effect, promoting neuronal survival and attenuating the motor deficit.

Acrobatic training prevents learning impairments and astrocyte remodeling in the hippocampus of rats undergoing chronic cerebral hypoperfusion: sex-specific benefits.

Background: Chronic cerebral hypoperfusion (CCH) leads to memory and learning impairments associated with degeneration and gliosis in the hippocampus. Treatment with physical exercise carries different therapeutic benefits for each sex. We investigated the effects of acrobatic training on astrocyte remodeling in the CA1 and CA3 subfields of the hippocampus and spatial memory impairment in male and female rats at different stages of the two-vessel occlusion (2VO) model. Methods: Wistar rats were randomly allocated into four groups of males and females: 2VO acrobatic, 2VO sedentary, sham acrobatic, and sham sedentary. The acrobatic training was performed for 4 weeks prior to the 2VO procedure. Brain samples were collected for morphological and biochemical analysis at 3 and 7 days after 2VO. The dorsal hippocampi were removed and prepared for Western blot quantification of Akt, p-Akt, COX IV, cleaved caspase-3, PARP, and GFAP. GFAP immunofluorescence was performed on slices of the hippocampus to count astrocytes and apply the Sholl's circle technique. The Morris water maze was run after 45 days of 2VO. Results: Acutely, the trained female rats showed increased PARP expression, and the 2VO-trained rats of both sexes presented increased GFAP levels in Western blot. Training, mainly in males, induced an increase in the number of astrocytes in the CA1 subfield. The 2VO rats presented branched astrocytes, while acrobatic training prevented branching. However, the 2VO-induced spatial memory impairment was partially prevented by the acrobatic training. Conclusion: Acrobatic training restricted the astrocytic remodeling caused by 2VO in the CA1 and CA3 subfields of the hippocampus. The improvement in spatial memory was associated with more organized glial scarring in the trained rats and better cell viability observed in females.

Coping with the experience of frustration throughout life: Sex- and age-specific effects of early life stress on the susceptibility to reward devaluation.

Early life stress may lead to lifelong impairments in psychophysiological functions, including emotional and reward systems. Unpredicted decrease in reward magnitude generates a negative emotional state (frustration) that may be involved with susceptibility to psychiatric disorders. We evaluated, in adolescents and adult rats of both sexes, whether maternal separation (MS) alters the ability to cope with an unexpected reduction of reward later in life. Litters of Wistar rats were divided into controls (non handled - NH) or subjected to MS. Animals were trained to find sugary cereal pellets; later the amount was reduced. Increased latency to reach the reward-associated area indicates higher inability to regulate frustration. The dorsal hippocampus (dHC) and basolateral amygdala (BLA) were evaluated for protein levels of NMDA receptor subunits (GluN2A/GluN2B), synaptophysin, PSD95, SNAP-25 and CRF1. We found that adult MS males had greater vulnerability to reward reduction, together with decreased GluN2A and increased GluN2B immunocontent in the dHC. MS females and adolescents did not differ from controls. We concluded that MS enhances the response to frustration in adult males. The change in the ratio of GluN2A and GluN2B subunits in dHC could be related to a stronger, more difficult to update memory of the aversive experience.

Influence of platform and abutment angulation on peri-implant bone. A three-dimensional finite element stress analysis.

The aim of this study was to evaluate stress distribution on the peri-implant bone, simulating the influence of Nobel Select implants with straight or angulated abutments on regular and switching platform in the anterior maxilla, by means of 3-dimensional finite element analysis. Four mathematical models of a central incisor supported by external hexagon implant (13 mm × 5 mm) were created varying the platform (R, regular or S, switching) and the abutments (S, straight or A, angulated 15°). The models were created by using Mimics 13 and Solid Works 2010 software programs. The numerical analysis was performed using ANSYS Workbench 10.0. Oblique forces (100 N) were applied to the palatine surface of the central incisor. The bone/implant interface was considered perfectly integrated. Maximum (σmax) and minimum (σmin) principal stress values were obtained. For the cortical bone the highest stress values (σmax) were observed in the RA (regular platform and angulated abutment, 51 MPa), followed by SA (platform switching and angulated abutment, 44.8 MPa), RS (regular platform and straight abutment, 38.6 MPa) and SS (platform switching and straight abutment, 36.5 MPa). For the trabecular bone, the highest stress values (σmax) were observed in the RA (6.55 MPa), followed by RS (5.88 MPa), SA (5.60 MPa), and SS (4.82 MPa). The regular platform generated higher stress in the cervical periimplant region on the cortical and trabecular bone than the platform switching, irrespective of the abutment used (straight or angulated).

Fluctuations in Seminal Quality throughout the Year: How do Temperature, Humidity and Atmospheric Pressure Impact on Human Sperm Quality?

Background: Most studies evaluating the possible seasonal variation of semen quality have considered temperature as the only causal factor. Aims: To assess possible seasonality in sperm quality and associations between semen parameters and several meteorological variables (temperature, humidity, apparent temperature and atmospheric pressure) in a large cohort of andrological patients. Settings and Design: This was a retrospective, cross-sectional and correlational/descriptive study. Materials and Methods: Patients (n: 15665) were categorised into four groups (summer, winter, spring and autumn) according to the date of assistance at the fertility centre. Daily values of temperature, apparent temperature, humidity and atmospheric pressure were provided by the National Weather System and were calculated as the average of the 74 days previous to semen collection (spermatogenic cycle). Statistical Analysis Used: As appropriate, the results were analysed by analysis of variance/Kruskal-Wallis, Chi-square test, t-test/Mann-Whitney, forward conditional regression model and Spearman/Pearson's correlations. Results: We detected seasonality effects on sperm count, total sperm count and total motile sperm count, with the highest values in winter and the lowest in summer. Correlation analysis showed that temperature, apparent temperature and humidity negatively correlated with semen parameters, being humidity the most powerful predictive meteorological variable. Conclusion: Sperm quality is influenced by seasons; increased environmental temperature and humidity negatively affect semen quality.

Bumetanide Attenuates Cognitive Deficits and Brain Damage in Rats Subjected to Hypoxia-Ischemia at Two Time Points of the Early Postnatal Period.

Neonatal hypoxia-ischemia (HI) is one of the main causes of tissue damage, cell death, and imbalance between neuronal excitation and inhibition and synaptic loss in newborns. GABA, the major inhibitory neurotransmitter of the central nervous system (CNS) in adults, is excitatory at the onset of neurodevelopment and its action depends on the chloride (Cl-) cotransporters NKCC1 (imports Cl-) and KCC2 (exports Cl-) expression. Under basal conditions, the NKCC1/KCC2 ratio decreases over neurodevelopment. Thus, changes in this ratio caused by HI may be related to neurological disorders. The present study evaluated the effects of bumetanide (NKCC cotransporters inhibitor) on HI impairments in two neurodevelopmental periods. Male Wistar rat pups, 3 (PND3) and 11 (PND11) days old, were submitted to the Rice-Vannucci model. Animals were divided into 3 groups: SHAM, HI-SAL, and HI-BUM, considering each age. Bumetanide was administered intraperitoneally at 1, 24, 48, and 72 h after HI. NKCC1, KCC2, PSD-95, and synaptophysin proteins were analyzed after the last injection by western blot. Negative geotaxis, righting reflex, open field, object recognition test, and Morris water maze task were performed to assess neurological reflexes, locomotion, and memory function. Tissue atrophy and cell death were evaluated by histology. Bumetanide prevented neurodevelopmental delay, hyperactivity, and declarative and spatial memory deficits. Furthermore, bumetanide reversed HI-induced brain tissue damage, reduced neuronal death and controlled GABAergic tone, maintained the NKCC1/KCC2 ratio, and synaptogenesis close to normality. Thereby, bumetanide appears to play an important therapeutic role in the CNS, protecting the animals against HI damage and improving functional performance.

Ghrelin misbalance affects mice embryo implantation and pregnancy success by uterine immune dysregulation and nitrosative stress.

Introduction: In a previous study we found that ghrelin (Ghrl) misbalance during the peri-implantation period significantly impaired fetus development. In this study we aimed to evaluate the putative mechanisms underlying these effects, including embryo implantation success, uterine nitric oxide synthase (NOS) activity, nitric oxide synthesis and the inflammatory/immune uterine profile. Methods: Ghrelin misbalance was induced by injecting 4nmol/animal/day of Ghrl (hyperghrelinemia) or 6nmol/animal/day of a Ghrl antagonist (Ant: (D-Lys3)GHRP-6) from day 3 to 8 of pregnancy. Control animals (C) were injected with de vehicle. Females were euthanized at pregnancy day 8 and their uteri excised in order to evaluate: the percentage of reabsorbed embryos (microscopically), eNOS, iNOS and nytrotirosine expression (by immunohistochemistry), nitrite synthesis (by Griess technique), VEGF, IL-10, IL-17, IL-6, MMP9 and GM-CSF expression (by qPCR) and leukocyte infiltration by flow cytometry (evaluating T cells, NK cells, granulocytes, dendritic cells and macrophages). Results: Ant-treatment significantly increased the percentage of reabsorbed embryos and the uterine expression of eNOS, iNOS and nytrotirosine. (D-Lys3)GHRP-6-treatment increased also the expression of the inflammatory cytokines IL-6, IL-17 and MMP9, and decreased that of IL-10 (anti-inflammatory). Moreover, Ant-treatment increased also the NK cells population and that of CD11b+ dendritic cells; and decreased T cells percentages. Similarly, hyperghrelinemia showed a significant increase vs. C on eNOS, iNOS and nytrotirosineuterine expression and a decrease in T cells percentages. Conclusion: Ghrl misbalance during the peri-implantation period induces pro-inflammatory changes and nitrosative stress in the gravid uterus, impairing significantly embryo implantation and/or development.

Straight and angulated abutments in platform switching: influence of loading on bone stress by three-dimensional finite element analysis.

PURPOSE: In view of reports in the literature on the benefits achieved with the use of platform switching, described as the use of an implant with a larger diameter than the abutment diameter, the goal being to prevent the (previously) normal bone loss down to the first thread that occurs around most implants, thus enhancing soft tissue aesthetics and stability and the need for implant inclination due to bone anatomy in some cases, the aim of this study was to evaluate bone stress distribution on peri-implant bone, by using three-dimensional finite element analysis to simulate the influence of implants with different abutment angulations (0 and 15 degrees) in platform switching. METHODS: Four mathematical models of an implant-supported central incisor were created with varying abutment angulations: straight abutment (S1 and S2) and angulated abutment at 15 degrees (A1 and A2), submitted to 2 loading conditions (100 N): S1 and A1-oblique loading (45 degrees) and S2 and A2-axial loading, parallel to the long axis of the implant. Maximum (σmax) and minimum (σmin) principal stress values were obtained for cortical and trabecular bone. RESULTS: Models S1 and A1 showed higher σmax in cortical and trabecular bone when compared with S2 and A2. The highest σmax values (in MPa) in the cortical bone were found in S1 (28.5), followed by A1 (25.7), S2 (11.6), and A2 (5.15). For the trabecular bone, the highest σmax values were found in S1 (7.53), followed by A1 (2.87), S2 (2.85), and A2 (1.47). CONCLUSIONS: Implants with straight abutments generated the highest stress values in bone. In addition, this effect was potentiated when the load was applied obliquely.