Protective effect of melatonin against methotrexate-induced testicular damage in the rat model: An experimental study

Background: Methotrexate (MTX) has been shown to affect the testes adversely, especially the seminiferous epithelium. As melatonin, an endocrine hormone, has been shown to normalize testicular function, its ability to prevent MTX-induced testicular damage should be considered. Objective: Based on the antioxidant, anti-inflammatory, and antiapoptotic activities of melatonin, this study aimed to investigate its protective effect against testicular damage induced by MTX. Materials and Methods: Forty adult male rats (200-230 g) were divided into five groups (n = 8/each). The rats in group I were injected with vehicle as a control. In group II, the rats were received intraperitoneal injections of melatonin (8 mg/kg) for 15 consecutive days. The rats in group III were intravenously injected with MTX (75 mg/kg) for 15 consecutive days. The remaining two groups received melatonin (8 mg/kgBW) for 15 (group IV) and 30 (group V) consecutive days, intraperitoneally, and then intravenously received MTX (75 mg/kgBW) on days 8 and 15 of the experimental period. Reproductive parameters, including epididymal sperm concentration, testicular tyrosine-phosphorylated protein expression, steroidogenic acute regulatory (StAR) protein expression, and caspase-3 and malondialdehyde levels, were examined. Results: The sperm concentrations (×106/ml) of groups IV (58.75 ± 1.28) and V (55.93 ± 2.57) were improved significantly (p = 0.032) compared with that of group II (32.92 ± 2.14). The seminiferous epithelium in groups IV and V also increased, while caspase- 3 expression decreased. In the melatonin-treated groups, the expression of tyrosinephosphorylated proteins at 32 kDa was decreased and that of proteins at 47 kDa was increased compared with the MTX group. StAR protein expression was not altered in any of the groups. Conclusion: Our results indicate that melatonin improves the epididymal sperm concentration by decreasing the expression of caspase-3 and increasing that of tyrosine-phosphorylated proteins in MTX-treated testes. Key words: Melatonin, Testis, Sperm, Methotrexate, Caspase-3, Tyrosine phosphorylation

Double inferior vena cava with three shunts: a rare anomaly with important implications for surgeons

Inferior vena cava (IVC) is the largest single vein that collects systemic venous blood from the lower part of the body except the gut and drains into the right atrium. Double IVC is a rare anomaly in humans and usually is discovered incidentally during the interventional radiological procedures or routine cadaveric dissection. Here we report a rare case of unusual observations in an adult female Thai cadaver with a duplicated left IVC with three short venous shunts and a variant pattern of the hemiazygos vein. Also included in this case was the presence of unilateral double renal vein on the right kidney. This type of anatomic variation of the great vein has never been reported before. A detailed description of these variations is useful and essential for the surgeons during approaching the retroperitoneal region

Effects of aged garlic extract on spatial memory and oxidative damage in the brain of amyloid-β induced rats

This study investigated the effect of aged garlic extract (AGE) on spatial learning and memory ability using Morris water maze (MWM) test in amyloid-β (Aβ) induced-neurotoxicity rats. Pretreatment of AGE at oral doses of 125, 250 and 500 mg/kg for 8 weeks significantly prevented the learning and short-term memory impairment in Aβ-induced neurotoxicity rats. Histological analysis has shown that pretreatment of AGE reversed the neuron loss in the CA1 and CA2 regions of hippocampus of Aβ-induced neurotoxicity in a comparable effect of ascorbic acid. By DPPH and FRAP determination, AGE had high antioxidative activity. Pretreatment of AGE caused significant increases of superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities, no significant change in catalase (CAT) activity, and a significant decrease of malondialdehyde (MDA) level of the Aβ-induced rat brain homogenate. The results suggest that AGE ameliorates the cognitive dysfunction in Aβ- induced neurotoxicity rats via its antioxidative effect

Evaluation of the Cariogram for root caries prediction

<p>The relative expression of plasma proteins at 24, 48 and 96 hpi of infected snails were compared against the control group (uninfected snails). Proteins were grouped into 7 different categories based on GO annotation and clustering was performed using Euclidean distances based on the expression patterns. Color intensity reflects the corresponding log₂ fold change and significant up-regulation shown in red, significant down-regulation is shown in blue, and no statistically significant change in abundance is shown in gray.</p

Asiatic Acid Prevents the Deleterious Effects of Valproic Acid on Cognition and Hippocampal Cell Proliferation and Survival

Valproic acid (VPA) is commonly prescribed as an anticonvulsant and mood stabilizer used in the treatment of epilepsy and bipolar disorder. A recent study has demonstrated that VPA reduces histone deacetylase (HDAC) activity, an action which is believed to contribute to the effects of VPA on neural stem cell proliferation and differentiation which may explain the cognitive impairments produced in rodents and patients. Asiatic acid is a triterpenoid derived from the medicinal plant Centella asiatica. Our previous study has shown that Asiatic acid improves working spatial memory and increases cell proliferation in the sub granular zone of the hippocampal dentate gyrus. In the present study we investigate the effects of Asiatic acid in preventing the memory and cellular effects of VPA. Male Spraque-Dawley rats were orally administered Asiatic acid (30 mg/kg/day) for 28 days, while VPA-treated animals received injections of VPA (300 mg/kg) twice a day from Day 15 to Day 28 for 14 days. Spatial memory was determined using the novel object location (NOL) test and hippocampal cell proliferation and survival was quantified by immuostaining for Ki-67 and Bromodeoxyuridine (BrdU), respectively. The results showed that VPA-treated animals were unable to discriminate between objects in familiar and novel locations. Moreover, VPA significantly reduced numbers of Ki-67 and BrdU positive cells. These results indicate that VPA treatment caused impairments of spatial working memory, cell proliferation and survival in the subgranular zone (SGZ) of the hippocampal dentate gyrus (DG). However, these abnormalities were restored to control levels by co-treatment with Asiatic acid. These data demonstrate that Asiatic acid could prevent the spatial memory and neurogenesis impairments caused by VPA

Fig 3. Effect of 5-FU and AA on proliferating cell counts in the dentate gyrus.

<p><b>Fig 3. Effect of 5-FU and AA on proliferating cell counts in the dentate gyrus. </b>Representative images of Ki-67 positive cells in the dentate gyrus in each group<b> (A-F)</b>. Ki-67 positive cells were stained green in the subgranular zone (SGZ) of the dentate gyrus. Section were counterstained with red nuclear dye, propidium iodide: PI. Arrowheads indicate Ki-67 positive cells in the the dentate gyrus (scale bars: 100 µm). Inserted images show Ki-67 immunostaining under high magnification (scale bar: 50 µm). Mean Ki-67 positive cell counts of the 5-FU group were significantly lower than the control, AA, and 5-FU+AA (preventive and throughout) groups (*<i>p</i><0.05, <b>G</b>). Inaddition, Ki-67 positive cell number in the 5-FU+AA (recovery) group were significantly lower than in the control group (#<i>p</i><0.05, <b>G</b>). ML: molecular layer, GCL: granule cell layer.</p

Data for: Protective effects of melatonin against valproic acid-induced memory impairments and reductions in adult rat hippocampal neurogenesis

Valproic acid (VPA) is widely used in the treatment of epilepsy. However, VPA has been revealed to impair memory in both humans and animals. The adverse effects of VPA are associated with reductions in hippocampal neurogenesis and memory. There are neuroprotective properties exerted by melatonin. As such, the present study investigated the protective effects of melatonin against the reductions of memory and neurogenesis caused by VPA. Male Spraque-Dawley rats received VPA (300 mg/kg) twice a day for 14 days, or melatonin (8 mg/kg/day) for 14 days, or co-treatment with VPA and melatonin for either 14 days (preventive and recovery groups) or 28 days (throughout group). Novel object location (NOL) and novel object recognition (NOR) tests were used to assess spatial memory and non-spatial memory, respectively. Proliferation, survival, and immature neurons in the subgranular zone (SGZ) were examined using Ki-67, BrdU and doublecortin (DCX) immunohistochemistry. Exploration time was scored when animals directed theirs nose toward the objects at a distance less than 2 cm. All sections were quantified at X40 on a Nikon ECLIPSE 80i fluorescence microscope. Ki-67, BrdU and DCX positive cells were considered within the SGZ, which is defined as 3 cell breadths of the internal rim of both blades of the dentate gyrus. The numbers of Ki-67, BrdU, and DCX positive cells in each section were calculated by accumulating cell counts per section for 9 sections per brain and multiplying the results by 15 for Ki67 and 8 for BrdU and DCX. The Student’s t-test and one-way ANOVA were used to analyse data where appropriate. Exploration times from both tests were calculated and converted to a preference index (PI), defined as time spent exploring for novelty in the choice trial as a percentage in comparison to 50% chance. The exploration time of either object placed in the novel location or novel object was significant greater than familiar location or object, demonstrating that the animals had normal cognition. If the PI is significant higher than 50% chance, suggesting that the animals did not have memory deficits. For immunostaining, the number of Ki-67, BrdU and DCX positive cells in control group was compared with valproic acid, melatonin, preventive, recovery, and throughout groups to show changes of neurogenesis

Fig 4. Effect of 5-FU and AA on cell survival in the dentate gyrus.

<p><b>Fig 4. Effect of 5-FU and AA on cell survival in the dentate gyrus.</b> Representative images of BrdU positive cells in the dentate gyrus in each group <b>(A-F)</b>. BrdU positive cells were stained green in the subgranular zone (SGZ) of the dentate gyrus. Sections were counterstained with red nuclear dye, propidium iodide: PI. Arrowheads indicate BrdU positive cells in the the dentate gyrus (scale bars: 100 µm). Inserted images show BrdU immunostaining under high magnification (scale bar: 50 µm). Mean BrdU positive cell counts in the 5-FU group were significantly lower than the control, AA, and 5-FU+AA (preventive and throughout) groups (*<i>p</i><0.05, <b>G</b>). Furthermore, BrdU positive cell counts in the 5-FU+AA (recovery) group were significantly less than those in the control group (#<i>p</i><0.05, <b>G</b>). ML: molecular layer, GCL: granule cell layer.</p