Effect of induced diabetes on morphometric indexes of the cerebellar cortex and gene expression in C57BL mice.

Objectives: Diabetes is a metabolic disorder that affects the development of the central nervous system and plays an important role in learning and memory. Diabetes increases the reactive oxygen species (ROS) level in cells and changes the expression of several genes, including SYP, BDNF, PAX7, and SYNCAM1, through the FOXO transcription factor. This study was done to assess the effect of diabetes on morphometric indexes of the cerebellar cortex and gene expression in mice. Materials and Methods: Diabetes was induced in twelve adult, male C57BL mice using an injection of streptozotocin. After two months, the mice were dissected, and the cerebellum was stored for further analysis. For the morphometric analysis, tissue sections were stained with cresyl violet and examined with a light microscope. For gene expression analysis, the RNA was extracted, and cDNA was synthesized. The mRNA levels of SYP, BDNF, PAX7, and SYNCAM1 genes were measured by the real-time PCR method. Results: The thickness of the molecular layer and Purkinje layer, and the number of Purkinje and granular cells in the diabetic group were significantly reduced compared to controls P<0.0 1). The area, perimeter, and diameter of Purkinje cells in the diabetic group were significantly reduced compared to controls P<0.0 1). The expression of PAX7, SYP, and BDNF genes of the diabetic group was significantly reduced. However, SYNCAM1 expression in the cerebellum of the diabetic group was significantly increased compared to controls (P<0.05). Conclusion: Induced diabetes in mice can decrease the expression of memory-related genes in the cerebellum. Also, these genes affect the morphology and thickness of the cerebellum.

Gestational diabetes mellitus increased the number of dopaminergic neurons in the olfactory bulb of rat offspring.

Objectives: Gestational Diabetes Mellitus (GDM) is the most common metabolic complication of pregnancy that causes central nervous system and olfactory dysfunction in the offspring. It has been demonstrated that dopamine modulates several aspects of olfactory information processing in vertebrates. Materials and Methods: In this study, we investigated the effect of gestational diabetes on the expression of the Dopamine (DA) metabolism genes, tyrosine hydroxylase (TH), and dopa decarboxylase (DDC) in the olfactory bulb (OB) tissue of rats' offspring. Female Wistar rats were divided into a control group which received citrate buffer and the diabetic group which received 45 mg/kg of streptozotocin (STZ) on day 0 of gestation. Fasting blood glucose levels were measured before and 72 hr after injection. OB tissues of adult offspring were isolated, and TH-positive cells were counted by immunofluorescence staining. Also, TH and DDC expressions were analyzed by qRT- PCR and western blot. Results: The data showed that gestational diabetes could cause up-regulation of TH (P<0.01) and DDC (P<0.05) in the OB tissue of offspring. Furthermore, our results showed that GDM causes a significant increase in TH and DDC protein levels in the OB tissues of offspring. Immunohistochemistry showed a significant increase in the number of TH-positive cells in the offspring of diabetic mothers (P<0.05). Conclusion: This study showed that gestational diabetes could cause an increase in TH and DDC gene expression in the OB tissue in the offspring, which may be correlated with reduced olfactory sensitivity.

Effect of MDMA exposure during pregnancy on cell apoptosis, astroglia, and microglia activity in rat offspring striatum.

Objectives: Ecstasy is a popular recreational psychostimulant with side effects on the central nervous system. This study examined the corpus striatum tissue of adult rats that received ecstasy during the embryonic period for histological and molecular studies. Materials and Methods: Rats were divided into control and ecstasy groups. The ecstasy group was given MDMA 15 mg/kg intraperitoneally twice daily at 8-hour intervals on days 7-15 of gestation. At the age of 15 weeks, adult offspring of both groups were examined for learning and memory study by the Morris water maze test. Then, ventral striatum tissue was harvested for TUNEL assay, Nissl staining, and real-time PCR for the expression of the GFAP and CD11b. Results: Ecstasy up-regulated the GFAP and CD11b expression in the striatum of offspring (*P˂0.05). Furthermore, the Morris water maze test showed that exposure to ecstasy significantly impaired learning and spatial memory (*P˂0.05). TUNEL assay results did not show any significant change in the number of apoptotic cells in the striatum tissue of ecstasy offspring compared with controls, while Nissl staining showed a significant decrease in the number of neurons in the ecstasy group (*P˂0.05). Conclusion: Exposure to ecstasy during pregnancy causes long-lasting changes in brain regions underlying learning and memory, including the striatum, and impaired working memory in the offspring. In addition, these data provide the first evidence that exposure to ecstasy during the embryonic period causes a persistent change in the activity of microglial cells and the number of astrocyte cells in the striatum.

In Utero Exposure to Gestational Diabetes Alters DNA Methylation and Gene Expression of CDKN2A/B in Langerhans Islets of Rat Offspring.

OBJECTIVE: DNA methylation, a major epigenetic reprogramming mechanism, contributes to the increased prevalence of type 2 diabetes mellitus (T2DM). Based on genome-wide association studies, polymorphisms in CDKN2A/B are associated with T2DM. Our previous studies showed that gestational diabetes mellitus (GDM) causes apoptosis in ß-cells, leading to a reduction in their number in pancreatic tissue of GDM-exposed adult rat offspring. The aim of this study was to examine the impact of intrauterine exposure to GDM on DNA methylation, mRNA transcription, as well as protein expression of these factors in the pancreatic islets of Wistar rat offspring. Our hypothesis was that the morphological changes seen in our previous study might have been caused by aberrant methylation and expression of CDKN2A/B. MATERIALS AND METHODS: In this experimental study, we delineated DNA methylation patterns, mRNA transcription and protein expression level of CDKN2A/B in the pancreatic islets of 15-week-old rat offspring of streptozotocin-induced GDM dams. We performed bisulfite sequencing to determine the DNA methylation patterns of CpGs in candidate promoter regions of CDKN2A/B. Furthermore, we compared the levels of mRNA transcripts as well as the cell cycle inhibitory proteins P15 and P16 in two groups by qPCR and western blotting, respectively. RESULTS: Our results demonstrated that hypomethylation of CpG sites in the vicinity of CDKN2A and CDKN2B genes is positively related to increased levels of CDKN2A/B mRNA and protein in islets of Langerhans in the GDM offspring. The average percentage of CDKN2A promoter methylation was significantly lower in GDM group compared to the controls (P<0.01). CONCLUSION: We postulate that GDM is likely to exert its adverse effects on pancreatic ß-cells of offspring through hypomethylation of the CDKN2A/B promoter. Abnormal methylation of these genes may have a link with ß-cell dysfunction and diabetes. These data potentially lead to a novel approach to the treatment of T2DM.

Gestational diabetes influences the expression of hypertrophic genes in left ventricle of rat's offspring.

OBJECTIVES: Gestational diabetes increases the risk of congenital heart disease in the offspring, but the molecular mechanism underlying this process remains unclear. Therefore, the current study was conducted to assess the effects of induced gestational diabetes on expression of some involved genes in cardiac hypertrophy in the offspring of diabetic rats. MATERIALS AND METHODS: Diabetes was induced in 40 adult Wistar rats by intraperitoneal injection of 45 mg/kg of streptozotocin. The day of appearance of the vaginal plug was assumed as day zero of gestation for inducing diabetes. After pregnancy, the offspring was maintained until they reach the age of 12 weeks. Then, their hearts were excised and were sectioned for molecular study. We analyzed the expression pattern of some hypertrophic genes by the quantitative real-time RT-PCR. RESULTS: The mRNA expression levels of all studied genes including c-jun, c-fos, c-myc, alpha-myosin heavy chain (α-MHC), atrial natriuretic factor (ANF) and ß-MHC, which are important in cardiomyocyte hypertrophy, were higher in the offspring of the diabetic group compared to controls. Significant differences were found for ß-MHC and c-myc with P<0.01 and for α-MHC and c-fos with P<0.05. CONCLUSION: Gestational diabetes upregulates expression of c-jun, c-fos c-myc, α-MHC, ANF and ß-MHC genes that are involved in cardiac hypertrophy in the offspring of diabetic rats.

Ethnicity Influences Corpus Callosum Dimensions.

BACKGROUND AND OBJECTIVE: Corpus callosum (CC), the main white matter cable which connects two hemispheres of brain, is important in special procedures such as stereotaxic surgeries vary in size, in different populations. Determination of possible size differences in ethnical groups has special values. PATIENTS AND METHODS: The size of the CC on midsagittal view was determined in 76 normal male subjects using MRI of brain hemispheres in northern Iran. The size of rostrum, body, splenium, length, and height of CC was measured for each subject. The width of the body of the corpus callosum (B), the anterior to posterior length (L) and the maximum height (H) of the corpus callosum, and ratios B/L and B/H were also calculated. RESULTS: The longitudinal dimensions of the CC were 70.21 mm and 74.05 mm in native Fars and Turkmens, respectively (P < 0.05). The heights were 25 mm and 25.75 mm in native Fars and Turkmen subjects, respectively. The width of CC in Turkmen people was significantly higher than native Fars people (P < 0.05). The Evans index in Turkmen group (0.314) was significantly higher than in native Fars (0.3). The B/L and B/H ratios were nonsignificantly different between two groups. CONCLUSION: The CC parameters vary in different ethnical groups in northern Iran.

Birth Defects in Northern Iran (2008-2013).

BACKGROUND: Congenital anomalies are important medical and public health conditions. The pattern and prevalence of birth defects may vary over time or with geographical location. We investigated the live birth prevalence and occurrence pattern of birth defects in Golestan Province, northern Iran. METHODS: This cross-sectional descriptive study was carried out on 144920 live newborns in 13 hospitals in Golestan Province, northern Iran, from 21 Jan 2008 to 19 Mar 2013. The newborns were examined for the presence of birth defects and mothers were interviewed for variables such as maternal age and ethnicity. In addition, data for each newborn was filed in a questionnaire and the coding of birth defects was translated to the International Classification of Diseases 10th revision-clinical modification (ICD-10-CM). RESULTS: Overall, 1690 infants were diagnosed as having birth defects among 144920 live newborns. The prevalence rate of birth defects was 11.66 per 1000 live births, the prevalence of birth defects per 1000 was11.62 in males and 11.42 in females. The prevalence of congenital anomalies among native Fars, Turkmen and Sistani were 13.03, 11.16 and 13.07, respectively, per 1000 live births. Anomalies of the cardiovascular system were the most common defects; the prevalence rate of cardiovascular system was 8.34 per 1000 live birth. CONCLUSION: The prevalence rate of birth defects in this area was lower than in the other regions in Iran (20.3 per 1000 live births) but higher than in some parts of Asia (7.33 per 1000 live births).

Exposure to mephedrone during gestation increases the risk of stillbirth and induces hippocampal neurotoxicity in mice offspring.

In recent years, abuse of synthetic cathinones, in particular, mephedrone, has increased among young adults worldwide. The study aim is to investigate the effects of mephedrone exposure during the gestational period on mice offspring outcomes, focusing on hippocampal neurotoxicity. The pregnant mice received mephedrone (50 mg/kg, sc) on a regular schedule (once daily on all days, from day 5 to 18 of gestation) or repeated schedule (thrice daily on day 5, 6, 11, 12, 17, and 18 of gestation) to simulate regular or recreational use of mephedrone, respectively. Results showed that the percentage of weight gain in pregnant mice was significantly lower in both regular and repeated schedule mephedrone groups (p < 0.01). Also, mephedrone significantly reduced the number and weight of delivered pups and increased the rate of stillbirth (p < 0.05). Immunohistochemistry and TUNEL assay showed an inhibition of cell proliferation (p < 0.05) and an increase of apoptosis (p < 0.05) in the hippocampus of delivered pups of the repeated schedule mephedrone group. This apoptotic effect was associated with enhanced expression of the pro-apoptotic Bax gene (p < 0.05) and reduction of expression of the anti-apoptotic Bcl-2 gene (p < 0.05) as evaluated by real-time PCR. The Morris water maze showed an impairment of spatial learning (p < 0.05) and reference memory (p < 0.01) in offspring born from litters exposed to mephedrone (repeated schedule). In conclusion, the present study has shown that regular and repeated exposure to mephedrone during the gestational period increases the risk of low birth weight and stillbirth. Also, repeated use of mephedrone impairs learning and memory processes through hippocampal damage.

Gestational diabetes leads to down-regulation of CDK4-pRB-E2F1 pathway genes in pancreatic islets of rat offspring.

OBJECTIVES: The link between a hyperglycemic intrauterine environment and the development of diabetes later in life has been observed in offspring exposed to gestational diabetes mellitus (GDM), but the underlying mechanisms for this phenomenon are still not clear. Reduced ß-cells mass is a determinant in the development of diabetes (type 1 and type 2 diabetes). Some recent studies have provided evidence that the CDK4-pRB-E2F1 regulatory pathway is involved in ß-cells proliferation. Therefore, we postulated that GDM exposure impacts the offspring's ß-cells by disruption in the CDK4-pRB-E2F1 pathway. MATERIALS AND METHODS: Adult Wistar rats were randomly allocated in control and diabetic group. The experimental group received 40 mg/kg/body weight of streptozotocin (STZ) on day zero of gestation. After delivery, diabetic offspring of GDM mothers and control dams at the age of 15 week were randomly scarified and pancreases were harvested. Langerhans islets of diabetic and control groups were digested by collagenase digestion technique. After RNA extraction, we investigated the expressions of the kir 6.2 and CDK4-pRB-E2F1 pathway genes by quantitative real-time PCR. RESULTS: GDM reduced the expression of CDK4-pRB-E2F1 pathway genes in Langerhans islets cells of offspring. CDK4, pRB and E2F1 pathway genes were downregulated in diabetic islets by 51%, 35% and 84%, respectively. Also, the expression of Kir 6.2 was significantly decreased in diabetic islets by 88%. CONCLUSION: We suggest that the effect of gestational diabetes on offspring's ß-cells may be primarily caused by the suppression of CDK4-pRB-E2F1 pathway.

Gestational diabetes influences retinal Muller cells in rat's offspring.

OBJECTIVES: The Muller cell is the principal glial cell of the vertebrate retina. The expression of Glial fibrillary acidic protein (GFAP) in the Muller cells was used as a cellular marker for retinal damage. This study was done to evaluate the effect of gestational diabetes on retinal Muller cells in rat's offspring. MATERIALS AND METHODS: In this experimental study, 12 Wistar rat dams were randomly allocated in control and diabetic groups. Gestational diabetes was induced by 40 mg/kg/body weight of streptozotocin at the first day of gestation, intraperitoneally. Dams in control group received an equivalent volume normal saline. Eye of six offspring of each group were removed at postnatal day 28 (P28). The histopathological changes in retina were examined through H&E staining and ultrastructure transmission electron microscopy (TEM). The expression of GFAP was examined using Immunohisto-chemical staining of GFAP in Muller cells. Photographs of retina were taken using Olympus BX51 microscope and a digital camera DP12 and EM LEO906; Zeiss, Germany. RESULTS: In the control rat's offspring, GFAP expression was not significant in Muller cells. According to the optical microscope images, GFAP expression was observed in the processes of the Muller cell in the inner plexiform layer of retina in offspring of diabetic mothers. In TEM technique, nuclear fragmentation and apoptotic bodies were observed in Muller cell of diabetic offspring. CONCLUSION: This study showed that the uncontrolled gestational diabetes can increase GFAP expression in Muller cells and retinal thickness of retinal layer in rat offspring's, therefore uncontrolled gestational can damage the Muller cells.

Gestational diabetes induces pancreatic beta-bells apoptosis in adult rat offspring / Diabetes gestacional induce apoptosis de células beta del páncreas en crías de ratas adultas

Several studies indicated that pancreatic ß-cell death occurs in both type 1 and type 2 diabetes. This experimental study was designed to determine the effect of gestational diabetes on the ß-cells in 16-week-old rat offspring. By this aim, adult Wistar rats aged 10-12 weeks were randomly allocated in control and diabetic groups. The diabetic group received 40 mg/kg/body weight of streptozotocin (STZ) on day zero of gestation. After delivery, diabetic offspring of GDM mothers and controls at the age of 16 weeks were sacrificed and pancreases harvested and fixed. The number of ß-cells and were counted by Gomori's method staining. Also, apoptosis in pancreas tissue of diabetic and control offspring was detected by TUNEL assay. Results showed a significant reduction in ß-cell number in offspring of GDM (p<0.05). TUNEL assay showed that the number of apoptotic cells increased in GDM compared to controls (P<0.05). This study revealed that gestational diabetes induces pancreatic beta-cells apoptosis in 16-week-old rat offspring.

Varios estudios indican que la muerte de las células ß del páncreas se produce tanto en la diabetes Tipo 1 como en la Tipo 2. Este estudio experimental fue diseñado para determinar el efecto de la diabetes gestacional en las células ß del páncreas en crías de ratas de 16 semanas. Para ello, ratas Wistar adultas de entre 10-12 semanas fueron asignadas al azar en dos grupos: control y diabetes. El grupo diabetes recibió 40 mg / kg / peso corporal de estreptozotocina (STZ) en el día cero de la gestación. Después del parto, a las 16 semanas, las crías de las madres diabéticas y controles de madres con diabetes gestacional (MDG), fueron sacrificadas para la extracción del páncreas, el cual posteriormente fue fijado. Se contó el número de células ß del páncreas mediante tinción con el método de Gomori. Además, se detectó apoptosis en el tejido del páncreas de la descendencia diabética y el grupo control mediante un ensayo TUNEL. Los resultados mostraron una reducción significativa en el número de células b en la descendencia de MDG (p <0,05). El ensayo TUNEL mostró que el número de células apoptóticas aumentó en MDG en comparación con los controles (P <0,05). Este estudio reveló que la diabetes gestacional induce apoptosis de células ß en el páncreas de crías de ratas de 16 semanas.

Gestational diabetes reduces pancreatic beta cells in rat offspring / La diabetes gestacional reduce las células beta pancreáticas en crías de ratas

Previous study has shown the adverse effects of gestational diabetes on hippocampal and spinal cord neuronal density in animal model. This study was conducted to determine the effect of gestational diabetes on beta cells in rat pancreas in early postnatal life. In this experimental study, 10 dams randomly allocated into control and diabetic groups on day 1 of gestation. Five dams in diabetic group received 40 mg/kg/BW of streptozotocin (intraperitoneally) and control animals received normal saline. Six of 28 and 56-day-old offspring of each gestational diabetes mellitus and controls were randomly scarified and sections were taken from the pancreas and stained using Gomorra's method. The density of beta cells and number and area of pancreatic islets were evaluated by quantitative computer-assisted morphometric method. The density of beta cells of 28-day-old offspring pancreas significantly reduced from 96.23±5.0 in control group to 71.5±5.3 cells in 10000 mm2 area of islet in diabetic group (P <0.01). The number of the pancreatic islets of in gestational diabetes (15.25±3.7) significantly reduced in comparison with the controls (8.61±0.7). The density of beta cells of 56-day-old offspring pancreas significantly reduced from 105.33±8.6 in control group to 62.12±5.9 in diabetic group (P <0.01). The number of the pancreatic islets of in gestational diabetes (13.5±0.5) significantly reduced compared to controls (6.75±1.7) (P <0.01). This study revealed that gestational diabetes loss the number of the beta cells in 28 and 56-day-old rat offspring.

Estudios previos han mostrado los efectos adversos de la diabetes gestacional en la densidad neuronal del hipocampo y de la médula espinal en modelos animales. Este estudio se llevó a cabo para determinar el efecto de la diabetes gestacional en las células beta del páncreas de rata en vida postnatal temprana. En este estudio experimental, 10 ratas fueron asignadas al azar a los grupos control y diabético en el día 1 de gestación. Cinco ratas del grupo diabético recibieron 40 mg/kg/BW de estreptozotocina (intraperitonealmente), mientras que los animales del grupo control recibieron solución salina normal. Seis de los descendientes, de 28 y 56 días de edad, de cada grupo, diabetes mellitus gestacional y control, se escarificaron al azar y se tomaron secciones del páncreas, que se tiñeron usando el método de Gomorra. La densidad de las células beta y el número y área de islotes pancreáticos fueron evaluados a través de método cuantitativo asistido por computadora morfométrica. La densidad de células beta del páncreas en las crías de 28 días disminuyó significativamente de 96,23 ± 5,0 en el grupo de control a 71,5 ± 5,3 células en el grupo diabético, en 10000 mm2 de área de islote (P <0,01). El número de islotes pancreáticos de la diabetes gestacional (15,25 ± 3,7) se redujo significativamente en comparación con los controles (8,61 ± 0,7). La densidad de células beta del páncreas en las crías de 56 días de edad se redujo de 105,33 ± 8,6 en el grupo de control a 62,12 ± 5,9 en el grupo diabético (P <0,01). El número de islotes pancreáticos en el grupo de diabetes gestacional (13,5 ± 0,5) se redujo significativamente en comparación con los controles (6,75 ± 1,7) (P <0,01). Este estudio reveló que la diabetes gestacional provoca una pérdida en el número de células beta en crías de ratas de 28 y 56 días de edad.

Ethnic difference in brain weight of 17-20 year-old males in Northern Iran / Diferencias étnicas en el peso del cerebro en hombres de 17 a 20 años de edad del norte de Irán

Cranial capacity and brain weight are important measurements in the study of racial/ethnic differences. Using linear (Lee­Pearson's) formula, brain weight and cranial capacity were estimated in 398 normal 17 to 20-year-old males (200 native Fars and 198 Turkman) males in Northern Iran. The dimensions of the head measured with spreading caliper and auricular head spanner. The mean±S.D of brain weight and cranial capacity in native Fars males were 1343.45±102.37 cm3, and 1390.47±105.95 g, and that of Turkmans were 1163.02±115.76 cm3 and 1203.73±119.81 g, respectively. Cerebral Index was 3.40 ±0.37 % and 2.52 ±0.37 % in Native Fars and Turkmans, respectively and cerebral quotient was higher in Turkmans (8.34) than Native Fars males (7.95). This study showed, the effect of ethnic factor influences the brain weight of 17-20 year-old males in Northern Iran.

La capacidad craneal y el peso del cerebro son mediciones importantes en el estudio de las diferentes etnias. Se estimó el peso del cerebro y la capacidad craneal usando la fórmula lineal de Lee-Pearson, en 398 hombres de 17 a 20 años de edad (200 nativos Fars y 198 Turcomanos) en el norte de Irán. Las dimensiones de la cabeza se midieron con caliper deslizante y con una llave de Todd, para la medición auricular. La media ± D.S. del peso del cerebro y la capacidad craneal en hombres nativos Fars fue de 1343,45±102,37 cm3, y 1390,47±105,95 g, y la de los turcomanos fue de 1163,02±115,76 cm3 y 1203,73 ± 119,81 g, respectivamente. El índice cerebral fue de 3,40± 0,37 % y 2,52 ± 0,37 % en nativos Fars y turcomanos, respectivamente, y el cociente cerebral fue mayor en los turcomanos (8,34) que en los hombres Fars (7.95). Este estudio demostró que el efecto del factor étnico influye en el peso del cerebro en hombres de 17-20 años del norte de Irán.

Hippocampal neuronal apoptosis in rat offspring due to gestational diabetes / Apoptosis neuronal del hipocampo en crías de ratas debido a la diabetes gestacional

Gestational diabetes mellitus (GDM) defined as impaired glucose tolerance affects approximately 6 % of all pregnant women who have never before had diabetes, but who do have high blood glucose levels during pregnancy. This study was done to evaluate the apoptosis in the neuronal cells in the CA1, CA2 and CA3 subfields of hippocampus and dentate gyrus in offspring of gestational diabetes at the 7, 21 and 28 d in postnatal rats. Thirty Wistar rat dams were randomly allocated in control and diabetic group. Dams in diabetic group were received 40 mg/kg/BW of streptozotocin at the first day of gestation and control groups received an equivalent volume normal saline injection intraperitoneally (IP). Six offspring of GDM and control dams, at the 7, 21, 28 postnatal day were randomly were sacrificed quickly with anesthesia. The coronal sections of brain serially collected. The apoptosis neurons were evaluated with TUNEL Assay. In the CA1, the number of apoptotic cells in 7, 21 and 28 d of postnatal life were significantly increased in GDM compared to controls (P<0.001). In the CA2, CA3 the number of apoptotic cells in 7, 21 and 28 d age-old offspring were significantly increased in GDM compared to controls (P<0.001). In the dentate gyrus, the number of apoptotic cells in 7, 21 and 28 d of postnatal life were significantly increased in GDM compared to controls (P<0.01). This study showed that the uncontrolled gestational diabetes significantly increases neuronal apoptosis in hippocampal and dentate gyrus in rat offspring.

La diabetes mellitus gestacional (DMG) se define como la intolerancia a la glucosa que afecta aproximadamente al 6 % de todas las mujeres embarazadas que nunca han tenido diabetes, pero que sí tienen niveles de glucosa en la sangre elevados durante el embarazo. El objetivo de este estudio fue evaluar la apoptosis de células neuronales en CA1, CA2 y CA3, subcampos del hipocampo y el giro dentado, en las crías de ratas con diabetes gestacional en los días 7, 21 y 28 luego del nacimiento. Se utilizaron 30 ratas Wistar asignadas aleatoriamente en grupos control y diabético (GDM). Se administró al grupo diabético 40 mg/kg de peso corporal de estreptozotocina en el primer día de gestación y el grupo control recibió un volumen equivalente de solución salina normal por inyección vía intraperitoneal. Seis crías de los grupos GDM y control fueron seleccionadas aleatoriamente y sacrificadas bajo anestesia los días 7, 21, 28. Se tomaron secciones seriales coronales del cerebro. La apoptosis neuronal se evaluó mediante ensayo TUNEL. En el CA1, el número de células apoptóticas a los 7, 21 y 28 d se incrementó significativamente en el grupo GDM en comparación con los controles (P <0.001). En el CA2 y CA3 el número de células apoptóticas en los días 7, 21 y 28 también se incrementó significativamente en GDM en comparación con los controles (P <0,001). En el giro dentado, el número de células apoptóticas en los días 7, 21 y 28 se incrementó significativamente en GDM en comparación con los controles (P <0,01). Este estudio mostró que la diabetes gestacional no controlada aumenta significativamente la apoptosis neuronal en el hipocampo y el giro dentado en las crías de las ratas.

Alterations of the giant pyramidal neurons (Betz cells) in brain cortex of rat offspring born from gestational diabetic dams: a morphometric study / Alteraciones de las neuronas piramidales gigantes de la corteza cerebral en crías de ratas nacidas de hembras con diabetes gestacional: un estudio morfométrico

A few studies reported the adverse effects of gestational diabetes on hippocampus and spinal cord of rat offspring. Giant pyramidal neurons are giant pyramidal neurons located in fifth layers of the gray matter in the primary motor cortex. Therefore, this study was conducted to determine the effect of gestational diabetes on the giant pyramidal neurons and the thickness of internal pyramidal layer in the brain cortex of rat offspring. On day 1 of gestation, 10 Wistar rat dams were randomly allocated into two control and diabetic groups. Five animals in diabetic group received 40 mg/kg/BW of Streptozotocin (intraperitoneally) and control animals received normal saline. We randomly selected six offspring of every subject in both groups at day 28, 56 and 84. Rat offspring were scarified and then coronal sections were taken from the right brain cortex and sections were stained with Cresyl violet. The density of giant pyramidal neurons in brain cortex and thickness of internal pyramidal layer of brain cortex were evaluated. In P28, P56, P84 the Betz cells density of brain cortex were significantly reduced from 107.6±6.2, 131.6±4.6 and 143.5±4.0 in controls to 84.96±2.1, 109.8±7.3 and 121.05±5.6 in cases (p<0.05), respectively. The thickness of the internal pyramidal layer of brain cortex in P28, 56 and P84 was significantly higher in gestational diabetic group in comparison with the control group (p<0.05). This study showed that uncontrolled gestational diabetes reduces the giant pyramidal neurons density and internal pyramidal layer thickness in brain cortex of rat offspring.

Pocos estudios han informado de los efectos adversos de la diabetes gestacional sobre las células del hipocampo y médula espinal. Este estudio, se realizó para determinar el efecto de la diabetes gestacional sobre las neuronas piramidales gigantes ubicadas en la quinta capas de la sustancia gris en la corteza motora primaria y el espesor de la capa piramidal interna en la corteza cerebral en crías de ratas. En el día 1 de la gestación, 10 ratas Wistar se asignaron aleatoriamente en dos grupos: control y diabéticos. Cinco animales del grupo diabético, fueron inyectados con 40 mg/kg de peso corporal de estreptozotocina (por vía intraperitoneal), y los de el grupo control, con solución salina. Aleatoriamente, se seleccionaron seis crías de cada hembra de ambos grupos los días 28, 56 y 84. Las crías fueron sacrificadas, se tomaron secciones coronales de la corteza cerebral derecha y se tiñeron con violeta de cresilo. Se evaluó la densidad de las neuronas piramidales gigantes en la corteza cerebral y el espesor de la capa piramidal interna de la corteza cerebral. En los días 28, 56, 84 la densidad de las neuronas piramidales gigantes en corteza cerebral se redujo significativamente al comparar los controles (107,6±6,2, 131,6±4,6 y 143,5±4,0 respectivamente) con los casos (84,96±2,1, 109,8±7,3 y 121,05±5,6 respectivamente) (p<0,05). El espesor de la capa piramidal interna de la corteza cerebral en los días 28, 84 y 56 fue significativamente mayor en el grupo diabético gestacional en comparación con el grupo control (p<0,05). Este estudio muestra que la diabetes gestacional no controlada reduce la densidad de neuronas piramidales gigantesy el espesor interno de la capa piramidal en la corteza cerebral de las crías de rata.

Effect of Gestational Diabetes on Purkinje and Granule Cells Distribution of the Rat Cerebellum in 21 and 28 days of Postnatal Life.

INTRODUCTION: Diabetes mellitus is associated with nervous system alterations in both human and animal models. This study was done to determine the effect of gestational diabetes on the Purkinje and granular cells in the cerebellum of rat offspring. METHODS: 10 Wistar rats Dams were randomly allocated in control and diabetic group. The experimental group received 40 mg/kg/body weight of streptozotocin (STZ) at the first day of gestation and control groups received saline injection intraperitoneally (IP). Six male offsprings of gestational diabetic mothers and control dams, at the 21, 28 postnatal days were randomly scarified and coronal sections of cerebellum (6 micrometer) serially collected. The neurons were stained with cresyl violet. RESULTS: The Purkinje cells density in the apex and depth of cerebellum in P21, in the experimental group was reduced 23% and 15% in comparison with the control group (P<0.001). The granular cells density in the experimental group was reduced 19.58% and 18.3% in comparison with the controls (P<0.001). The Purkinje cells density of cerebellum in P28, in the diabetic group reduced to 22.12% and 12.62% in comparison with the control group (P<0.001). The granular cells density in the diabetic group reduced 17.14% and 16.12% in comparison with the control group (P<0.001). DISCUSSION: The Purkinje and granular cells significantly reduced in gestational diabetes rat offspring.

Ganglionic cells apoptosis in retinal layer of rat offspring due to gestational diabetes / Células ganglionares apoptóticas en la capa de la retina de crías de ratas debido a la diabetes gestacional

Previous studies have shown the adverse effects of gestational diabetes on hippocampal neuronal density in animal models. This study was conducted to determine the effect of gestational diabetes on retinal ganglionic cell density, the thickness of the retinal layer and apoptotic ganglionic cell density in 28-day-old of rat offspring. In this experimental study, 10 Wistar rat dams were randomly allocated in control and diabetic groups. Gestational diabetes was induced by 40 mg/kg/body weight of streptozotocin at the first day of gestation, intraperitoneally, dams in control group received an equivalent volume normal saline. At postnatal day 28, six offspring of each gestational diabetes and controls were randomly selected, sacrificed and sections (6 micrometer) were taken from the eye and stained with hematoxylin and eosin. The density of ganglionic cells and the number of dUTP end-labeling (TUNEL)-positive cells were evaluated in 20000 mm2 area of ganglion layer of the retina. The ganglionic cells density were reduced (27.4%) in gestational diabetic offspring in compared to controls (22.5±1.5 vs. 31.0±0.9, P<0.01). The apoptotic ganglionic cells of retina in interventional group significantly increased in compared to controls (6.74±0.60 vs. 5.12±0.26, P<0.02). This study showed that the uncontrolled gestational diabetes can reduce the number of ganglionic cells and increase apoptotic ganglionic cells of retina layer in rat offspring.

Estudios previos en un modelo animal han demostrado los efectos adversos de la diabetes gestacional en la densidad neuronal del hipocampo. El objetivo fue determinar el efecto de la diabetes gestacional en la densidad de las células ganglionares de la retina, en el espesor de la capa de la retina y en la densidad de las células apoptóticas ganglionares, en crías de ratas de 28 días. En este estudio experimental, 10 ratas Wistar fueron asignadas aleatoriamente en grupos control y diabéticos. La diabetes gestacional se indujo a partir de la administración de 40 mg/kg/peso corporal de estreptozotocina en el primer día de la gestación, por vía intraperitoneal. Al grupo control se administró un volumen equivalente de solución salina normal. En el día 28 luego del nacimiento, se seleccionaron aleatoriamente seis crías procedentes de los grupos con diabetes gestacional y controles, se eutanasiaron y se tomaron muestras de los ojos, en forma de secciones de 6 micrómetros, las cuales se tiñeron con H & E. La densidad de las células ganglionares y el número final de células dUTP positivas (TUNEL) se evaluaron a nivel de la capa ganglionar de la retina, en un área de 20.000 mm2. La densidad de las células ganglionares se redujo un 27,4% en la descendencia con diabetes gestacional en comparación con los controles (22,5±1,5 vs. 31,0±0,9, P<0,01). Las células ganglionares apoptóticas de la retina en el grupo con diabetes gestacional aumentaron significativamente en comparación con los controles (6,74±0,60 vs. 5,12 ± 0,26, P <0,02). Este estudio demostró que la diabetes gestacional no controlada puede reducir el número de células ganglionares y aumentar el número de células ganglionares apoptóticas de la capa de la retina en las crías de las ratas con diabetes gestacional.

Neonatal and prenatal causes of death in Gorgan-North of Iran.

BACKGROUND: Reduction of neonatal mortality rate can improve health and newborn status of the society. This study was done to evaluate the prevalence and pattern of causes for neonatal mortality in Gorgan, North of Iran. METHODS: this descriptive, cross-sectional study was carried out on all birth in the maternity Dezyani hospital in Gorgan, North of Iran during 1 September 2008 -31 March 2011. Causes of mortality were evaluated in newborns that admitted and died in the NICU. Died newborns were screened for primary and final causes of death. The final causes of the mortality in majority of neonates were extracted according to the International Coding of disease Ver10 (ICD10). RESULTS: The total number of births was14785. The leading primary obstetric causes of death were: spontaneous preterm labor 36(25.7%), fetal abnormality 31(22.1%), hypertensive disorders 21(15%), unexplained intrauterine death 21(15%), maternal disease 12 (8.6%), intra-uterine growth restriction (IUGR) 10 (7.1%), oligohydroamnios 5(3.6%) and ante partum hemorrhage 4 (2.9%). Mortality rate was 76.8% during the first week of life. General final causes of death were; pulmonary bleeding17.9%, septic shock16.8%, IVH15.8%, congenital anomalies and pnomotorax 13.7% in fourth grade. CONCLUSION: The risk factors, main associated reason and procedures for prevention of spontaneous preterm labor and fetal abnormality should be includes as the main themes in perinatal research. In addition, there should be emphasized on necessity of genetic consultations and health care before and during pregnancy.

Reduction in non syndromic oral clefts following mandatory flour fortification with folic acid in Northern Iran.

UNLABELLED: Background Several studies have examined the effects of folic acid fortification on the occurrence of orofacial clefts. This study was carried out to evaluate the impact of flour fortification with folic acid on oral clefts in Northern Iran. METHODS: In this hospital based study from March 2006 to September 2009, we screened the total number of live births at all hospitals in Golestan province in North of Iran, as well as the number of pregnancies affected by non syndromic oral clefts, per 100000 births during the periods before (March 2006 -June 2007), during (July2007- March 2008) and after (May 2008- September 2009) folic acid fortification of flour was implemented. Non syndromic Oral clefts were defined in accordance with the International Classification of Diseases, 10th revision (ICD-10). RESULTS: The prevalence of oral clefting reduced from 64.2 per 100000 births before fortification to 32.2 per 100000 after flour fortification with folic acid. The preventive fraction rate of oral clefting after fortification period in comparison with before period was 49.8% (OR=0.502 95% CI 0.26-0.97, p=0.036). CONCLUSION: This study showed that flour fortification with folic acid is associated with a significant reduction in the rate of oral clefting in Golestan province in Northern Iran.

Risk factors of neural tube defects in northern iran.

BACKGROUND: Neural tube defects (NTDs) including spina bifida and anencephaly are the second most common birth defects with 2.8 per 1000 births in northern Iran. OBJECTIVES: This study was conducted to determine the risk factors of neural tube defects in Gorgan, north of Iran. PATIENTS AND METHODS: This hospital-based, case-control study was carried out on all NTD-affected pregnancies (n = 59) during February 2007 - August 2010, and 160 healthy pregnancies were selected via convenient sampling method in three hospitals in Gorgan, north of Iran. Risk factors including maternal body mass index (BMI), season of birth, gender of the newborn, mother's age, ethnicity, consanguineous marriage, folic acid consumption, nutrition, habitat, and education, were assessed through interviews with mothers. Univariate and multivariate logistic regression analyses were used to estimate the risks by odds ratios (ORs) and 95% confidence intervals. RESULTS: THE MULTIVARIATE ANALYSIS SHOWED THAT MATERNAL BMI (NORMAL/UNDERWEIGHT OR: 0.23, overweight/underweight OR: 0.15, obese/underweight OR: 0.13) and maternal ethnicity (Fars/Sistani OR: 3.49) and maternal nutrition (good/poor OR: 0.46) were significantly correlated with NTDs in the newborns. CONCLUSIONS: This study showed that maternal ethnicity, insufficient nutrition, and BMI, were the main risk factors of NTDs in northern Iran.