Experimental study of pre- and postnatal caffeine exposure and its observable effects on selected neurotransmitters and behavioural attributes at puberty : Caffeine exposure and its observable effects on selected neurotranmitters and behaviour.

Caffeine is globally consumed as a stimulant in beverages. It is also ingested in purified forms as power and tablets. Concerns have been raised about the potential consequences of intrauterine and early life caffeine exposure on brain health. This study modeled caffeine exposure during pregnancy and early postanal life until puberty, and the potential consequences. Caffeine powder was dissolved in distilled water. Thirty-two (n = 32) pregnant mice (Mus musculus) (dams) were divided into four groups- A, B, C and D. Group A animals served as a control, receiving placebo. Caffeine doses in mg/kg body weight were administered as follows: Group B, 10 mg/kg; Group C, 50 mg/kg; Group D, 120 mg/kg. Prenatal caffeine exposure [phase I] lasted throughout pregnancy. Half the number of offspring (pups) were sacrificed at birth; the rest were recruited into phase II and the experiment continued till day 35, marking puberty. Brain samples were processed following sacrifice. γ-aminobutyric acid (GABA), acetylcholine (ACh), and serotonin (5Ht) neurotransmitters were assayed in homogenates to evaluate functional neurochemistry. Anxiety and memory as neurobehavioural attributes were observed using the elevated plus and Barnes' mazes respectively. Continuous caffeine exposure produced positive effects on short and long-term memory parameters; the pattern interestingly was irregular and appeared more effective with the lowest experimental dose. Anxiety test results showed no attributable significant aberrations. Caffeine exposure persistently altered the neurochemistry of selected neurotransmitters including ACh and 5Ht, including when exposure lasted only during pregnancy. ACh significantly increased in group BC+ to 0.3475µgg-1 relative to control's 0.2508µgg-1; pre-and continuous postnatal exposure in Group B increased 5Ht to 0.2203 µgg-1 and 0.2213 µgg-1 respectively relative to control's 0.1863 µgg-1. From the current investigation, caffeine exposure in pregnancy had persistent effects on brain functional attributes including neurotransmitters activities, memory and anxiety. Caffeine in moderate doses affected memory positively but produced negative effects at the higher dosage including increased anxiety tendencies.

Neural and behavioural changes in male periadolescent mice after prolonged nicotine-MDMA treatment.

The interaction between MDMA and Nicotine affects multiple brain centres and neurotransmitter systems (serotonin, dopamine and glutamate) involved in motor coordination and cognition. In this study, we have elucidated the effect of prolonged (10 days) MDMA, Nicotine and a combined Nicotine-MDMA treatment on motor-cognitive neural functions. In addition, we have shown the correlation between the observed behavioural change and neural structural changes induced by these treatments in BALB/c mice. We observed that MDMA (2 mg/Kg body weight; subcutaneous) induced a decline in motor function, while Nicotine (2 mg/Kg body weight; subcutaneous) improved motor function in male periadolescent mice. In combined treatment, Nicotine reduced the motor function decline observed in MDMA treatment, thus no significant change in motor function for the combined treatment versus the control. Nicotine or MDMA treatment reduced memory function and altered hippocampal structure. Similarly, a combined Nicotine-MDMA treatment reduced memory function when compared with the control. Ultimately, the metabolic and structural changes in these neural systems were seen to vary for the various forms of treatment. It is noteworthy to mention that a combined treatment increased the rate of lipid peroxidation in brain tissue.

Stress-Sensitive Regulators of Fetal Neurodevelopment in HIV and Preeclampsia: An Immunocytochemical Appraisal of Placental OGT and T4 Levels.

Preeclampsia and HIV are a significant burden to maternal health globally, especially in low-middle income countries such as South Africa. In the KwaZulu-Natal province, SA antenatal HIV prevalence is 41.1%, while PE is 12%. PE and HIV infections are maternal stress and inflammation that impact placental function and fetal development. Therefore, this study investigated the impact of the comorbidity of PE and HIV on placental stress and neurodevelopment. Placentae were obtained from four cohorts of pregnant women: normotensive HIV negative, normotensive HIV positive, preeclamptic HIV negative, and preeclamptic HIV positive. The placental tissue sections were immunostained for OGT and T4. Our findings showed that the maternal weight, diastolic, and systolic blood pressures (BP) were higher in PE vs. the normotensive groups, irrespective of HIV status. In addition, significant changes were noticed in the placental weight, fetoplacental ratio, and placental efficiency coefficient. Our findings showed that the maternal weight, diastolic, and systolic blood pressures (BP) were statistically higher in the PE compared to the normotensive. No significant differences were observed between HIV positive and HIV negative groups. In addition, significant changes were noticed in the placental weight, fetoplacental ratio, and placental coefficient. Furthermore, considerable upregulation in the placental expression of OGT in both the conducting and exchange villi of PE and concomitant downregulation in HIV-positive patients compared with Normotensive and HIV-negative individuals, respectively. Our results provide inferential evidence on the dysregulation of OGT in the comorbidity of PE and HIV. This may mediate a compromised programmed outcome of an adverse maternal environment during pregnancy and consequently affect fetal development.

Variable Cre Recombination Efficiency in Placentas of Cyp19-Cre ROSAmT/mG Transgenic Mice.

The aromatase-Cre recombinase (Cyp19-Cre) transgenic mouse model has been extensively used for placenta-specific gene inactivation. In a pilot study, we observed unexpected phenotypes using this mouse strain, which prompted an extensive characterization of Cyp19-Cre placental phenotypes using ROSAmT/mG transgenic reporter mice. The two strains were mated to generate bi-transgenic Cyp19-Cre;ROSAmT/mG mice following a standard transgenic breeding scheme, and placental and fetal tissues were analyzed on embryonic day 17.5. Both maternal and paternal Cre inheritance were analyzed by mating the respective Cyp19-Cre and ROSAmT/mG males and females. The genotype results showed the expected percentage of Cyp19-Cre;ROSAmT/mG fetuses (73%) and Cre mRNA was expressed in all of the Cyp19-Cre placentas. However, surprisingly, only about 50% of the Cyp19-Cre;ROSAmT/mG placentas showed Cre-mediated recombinase activity as demonstrated by placental enhanced green fluorescent protein (EGFP) expression. Further genetic excision analysis of the placentas revealed consistent results showing the absence of excision of the tdTomato in all of the Cyp19-Cre;ROSAmT/mG placentas lacking EGFP expression. Moreover, among the EGFP-expressing placentas, there was wide variability in recombination efficiency, even in placentas from the same litter, leading to a mosaic pattern of EGFP expression in different zones and cell types of the placentas. In addition, we observed a significantly higher percentage of Cre recombination activity in placentas with maternal Cre inheritance. Our results show frequent mosaicism, inconsistent recombination activity, and parent-of-origin effects in placentas from Cyp19-Cre;ROSAmT/mG mice, suggesting that tail-biopsy genotype results may not necessarily indicate the excision of floxed genes in Cyp19-Cre positive placentas. Thus, placenta-specific mutagenesis studies using the Cyp19-Cre model require extensive characterization and careful interpretation of the placental phenotypes for each floxed allele.

Knowledge, attitudes and behaviours toward COVID-19: A cross-sectional survey among Nigerian University students.

Background: The effect of the highly contagious coronavirus disease 2019 (COVID-19) began in Wuhan, Hubei Province, China, from which it spread worldwide. In Nigeria, to curb the spread of the virus, the government elected to close public places, halt the general use of public transportation, enforce isolation and manage infected persons. Aim: This study evaluated Nigerian university students' knowledge, attitudes and behaviour (KAB) towards COVID-19. Setting: This was an online survey of Nigerian university students. Method: A cross-sectional study was conducted among 1268 respondents aged 16 to 60 who completed the survey questionnaire. The respondents' demographic data and KAB toward COVID-19 were collected, allocated and scored based on specific stratified divisions. Data were analysed using student's t-test, analysis of variance and logistic regression analysis. Results: The respondents demonstrated good knowledge of COVID-19, with a mean knowledge score of 78.7%; this positively influenced their attitude and behaviour scores (84.1% and 72.3%, respectively). Multiple linear regression analysis showed that 98.9% of the variance associated with poor knowledge is explained by gender (98.9%), age (97.3%), education (97.3%), occupation (97.2%) and marital status (91.4%). Conclusion: The respondents had a positive attitude and satisfactory compliance with safety practices required to curb the spread of the virus. Nevertheless, there is a need to intensify health education campaigns targeting all Nigerians, especially the less educated, via community outreach programmes using local languages. Contribution: The findings of this study demonstrate the imperative role of the knowledge of COVID-19 in curbing the spread of the infection via improved attitudes and positive behaviours in compliance with safety practices.

Differential upregulations of SMAC and LAMIN B levels in the buffy coat of HIV associated preeclamptic women.

OBJECTIVE: To assess HIV positivity as an apoptotic confounding variable in pregnancies complicated by preeclampsia. METHODS AND MATERIALS: Using a Bio-plex Multiplex Immunoassay, Smac and Lamin B concentrations (ng/ml) were analyzed in a buffy coat collected from 128 pregnant women attending a large regional hospital in Durban, South Africa. Study groups consisted of Normotensive and Preeclamptic pregnant women stratified according to their HIV status. All HIV positive groups received highly active antiretroviral therapy (HAART). RESULTS: Our findings showed significant (p < .05) upregulation in the levels of both SMAC and LAMIN B in the HIV positive patients and a concomitant downregulation of the same apoptotic makers were observed in preeclampsia regardless of HIV status. CONCLUSIONS: These results could be associated with the fact that apoptosis promotes deregulation of mitochondrial dynamics, contributing to the associated severe obstetric events observed in pregnancies among HIV-infected women on HAART.

Early glutathione intervention educed positive correlation between VGLUT1 expression and spatial memory in the Nω-nitro-L-arginine methyl rat model of IUGR.

INTRODUCTION: One of the most compelling causes of perinatal mortality and morbidity is intrauterine growth restriction (IUGR). IUGR is linked with numerous health challenges that last lifelong, including neurodevelopmental impairment and a high incidence of brain dysfunction. There is mounting evidence that places the glutamatergic system at the center of the neurobiology and treatment of neurological diseases. Therefore, this study investigated the effects of postnatal glutathione intervention on the spatial memory and the expressions of vesicular glutamate transporter 1 (VGLUT1) in the hippocampus and the cerebellar cortex of Nω-nitro-L-arginine methyl (L-NAME)-induced rat model of IUGR. MATERIALS AND METHOD: Twelve adult female rats were divided into Control and L-NAME groups; each containing 6 female rats. The control group received a single daily dose of normal saline while the L-NAME group was administered 50 mg/kg L-NAME daily from gestational day 9 until parturition. Offspring of the control rats were given free access to feeds while offspring from the L-NAME group were assigned into 3 groups: G1: given free access to feeds; G2 and G3 were administered 1.5 mg/kg body weight of glutathione from postnatal day (PND) 4-9 and PND 25-31 respectively. At the end of the intervention, Y-maze was conducted, and the rats euthanized on PND 35. The brain sections were processed, and immunofluorescence staining was performed using the Vectafluor Excel R.T.U Antibody kit. RESULTS: IUGR caused a significant 31.1% decrease in spontaneous alternation percentage (SAP), while early treatment with glutathione at PND 4-9 significantly (p < 0.01) increased SAP, while late treatment at PND 25-9 significantly decreased SAP compared to IUGR group. Furthermore, IUGR caused significant (p < 0.001) downregulation in corrected total cell fluorescence (CTCF) of VGLUT1 in both the hippocampus and cerebellar cortex. While treatment with glutathione caused upregulation in CTCF of VGLUT1 in the hippocampus and the cerebellar cortex. CONCLUSION: Our results showed that early intervention with glutathione has significant therapeutic potential via upregulation of VGLUT1 expression in both hippocampus and cerebellar cortex, which positively correlated with enhanced spatial memory in IUGR rat model.

VEGF Maintains Maternal Vascular Space Homeostasis in the Mouse Placenta through Modulation of Trophoblast Giant Cell Functions.

Vascular endothelial growth factor (VEGF) is an angiogenic growth factor that acts primarily on endothelial cells, but numerous studies suggest that VEGF also acts on non-endothelial cells, including trophoblast cells. Inhibition of VEGF signaling by excess production of the endogenous soluble VEGF receptor sFlt1 in trophoblast cells has been implicated in several pregnancy complications. Our previous studies and other reports have shown that VEGF directly regulates placental vascular development and functions and that excess VEGF production adversely affects placental vascular development. Trophoblast giant cells (TGCs) line the maternal side of the placental vasculature in mice and function like endothelial cells. In this study, we specifically examined the effect of excess VEGF signaling on TGC development associated with defective placental vascular development using two mouse models an endometrial VEGF overexpression model and a placenta-specific sFlt1 knockdown model. Placentas of endometrial VEGF-overexpressing dams at embryonic days (E) 11.5 and 14.5 showed dramatic enlargement of the venous maternal spaces in junctional zones. The size and number of the parietal TGCs that line these venous spaces in the placenta were also significantly increased. Although junctional zone venous blood spaces from control and VEGF-overexpressing dams were not markedly different in size at E17.5, the number and size of P-TGCs were both significantly increased in the placentas from VEGF-overexpressing dams. In sFlt1 knockdown placentas, however, there was a significant increase in the size of the sinusoidal TGC-lined, alkaline phosphatase-positive maternal blood spaces in the labyrinth. These results suggest that VEGF signaling plays an important role in maintaining the homeostasis of the maternal vascular space in the mouse placenta through modulation of TGC development and differentiation, similar to the effect of VEGF on endothelial cells in other vascular beds.

Comparative study on the influence of fluoride on lipid peroxidation and antioxidants levels in the different brain regions of well-fed and protein undernourished rats.

Effects of fluoride on the levels of Lipid peroxidation (LP) and antioxidant enzymes in the brain regions of protein undernourished (PU) and well-fed rats (WF) rats exposed to 100 ppm fluoride in drinking water were investigated. The results indicate that the mean body weights and the total brain weights of PU rats as well as those given fluoride (both WF and PU) were significantly (P < 0.05) lower than their respective controls. The weights of different brain regions were also significantly reduced (P < 0.05) in PU rats compared to WF rats except in the brain stem. Fluoride ingestion diminished the weights of WF and PU rats affecting the cerebrum only (in the case of PU rats) and the cerebellum of both WF and PU rats without an effect on the brain stem of both WF and PU. Additionally, increased LP was observed in the cerebrum and cerebellum of PU rats but after fluoride ingestion, 30% increase in LP was observed only in the cerebrum. In the brain stem however, protein undernutrition was accompanied with a significant reduction in LP but the region seems insensitive to fluoride. There were significant reductions (P < 0.05) in CAT, SOD and GSH in all the brain regions (except the GSH level in the brain stem only) of PU rats. Fluoride induced reduction in the activity of CAT in the three brain regions and on SOD activity in cerebrum only for WF rats but no effect of fluoride on all the antioxidants studied in the three brain regions for PU rats. It is concluded that WF and PU rats responded differently to fluoride toxicity. However, it seems that at the dosage used, fluoride toxicity may be a direct effect on the antioxidant enzymes.