Correlation studies on pathological changes in brain with neurotransmitters and behavioural changes in Balb/c mice

Aim: To determine the plasma neurotransmitters simultaneously and to find any correlation with pathological changes in the hippocampus and Purkinje cells and their relation with behavioral changes in Balb/c mice. Methodology: In the present study, both sexes of Balb /C mice were divided into two groups (4 males and 4 females; n = 8): Both the groups were given a single dose of either saline or sodium valproate (400mg kg-1) respectively through subcutaneous injection on PND 14. Behavioural tests were conducted on mice pups on various postnatal days till 40th day. On PND 41, blood samples were collected from all the animals for quantification of the neurotransmitters (serotonin, dopamine, and noradrenalin) in plasma, animals were sacrificed by cervical dislocation and whole brain was isolated for histological examination of the Purkinje cells and hippocampus. Results: Sodium valproate exposed animals showed loss of motor skill development (delayed negative geotaxic response), increased locomotor activity, increased anxiety, and retardation in water maze performance, and lower social interaction. Histopathological evolutions of cerebellum purkinje cells and hippocampus showed 40-50% atrophic cells in sodium valproate animals compared to control animals. Interpretation: The results of the present study indicate that Sodium valproate changes specific brain cell population in Balb/C mice, which might be the reason for the altered neurotransmitter levels, leading to behavioural changes in these animals.

Phosphate-buffered Formalin Fixative Under Controlled Fixation Temperature Yielded Excellently Preserved Histomorphology

Aim: Using mouse liver as experimental model, this study attempts to identify a formalin-based fixative and fixation temperature that jointly provides the best balance of preservation of tissue morphology. Methodology: Liver samples from fifty (50) albino mice aged between of 6 to 8 weeks consisting of both male and female was harvested following cervical dislocation and randomly distributed into control and experimental groups. Control samples were fixed in 10mL of 10% formalin at 25oC, 30oC, 35oC, 40oC, 45oC, 50oC, 55oC and 60oC respectively for 24 hours, while experimental samples were each fixed in equal volume of phosphate-buffered 10% formalin (pH 7.2, 7.4, 7.6 and 7.8) at the same temperature and time duration regimen and processed for general tissue morphology. Nuclear, cytoplasm and cell membrane morphology were assessed as evidence of the combined effectiveness of fixative and fixation temperature. Morphology was scored using a four-point grading scale with 1 being poor and 4 being excellent. Results: Nuclear, cytoplasm and cell membrane morphology were excellently preserved in tissue fixed with phosphate-buffered 10% formalin (pH 7.2) at 45oC. Tissue fixed with 10% formalin at 35oC exhibited excellent nuclear and cell membrane morphology, while excellent preservation of cell membrane morphology were observed in tissues fixed with 10% formalin at 40oC, phosphate-buffered 10% formalin (pH 7.4) at 55oC and 60oC, (pH 7.6) at 50oC and 55oC and (pH 7.8) at 55oC respectively. Furthermore, excellent preservation of nuclear morphology was observed in tissue fixed with phosphate-buffered 10% formalin (pH 7.8) at 60oC. Conclusion: Phosphate-buffered 10% formalin at a temperature of 45oC and pH 7.2 provide an excellent formalin-based fixative and fixation temperature that adequately preserves the microanatomy of tissue for histopathology examination.

The Changes of Prostatic Microvessel Density in Sprague-Dawleyrats After Castration Under the Effect of Estrogen/ Androgen at Different Concentrations

Background: Currently, there are relatively few studies on the effects of changes in estrogen and androgen levels on prostatic MVD.This article aimed to study the changes of prostatic MVD in SD rats after castration under the effect of estrogen/androgen at different concentrations.Methods: Male Sprague-Dawley(SD) rats aged 3-4 months were randomly divided into the control group, castration group, and different concentrations of estrogen/ androgen treatment after castration. Dihydrotestosterone(DHT) and estradiol(E) were administered daily by subcutaneous injection for one month. All the rats were sacrificed by cervical dislocation after one month, and the serum DHT and E concentrations of the rats in each group were measured by ELISA assay. Prostate tissues specimens were immunohistochemically stained with monoclonal antibodies against CD-34 and factor VIII for the MVD.Results: Compared with the control group, the MVD decreased significantly in the castration group (P<0.05). When the exogenous E concentration was constant, in general, the MVD of rats in all the groups increased with the increase of exogenous DHT concentration; Among them, compared with the castration group, the MVD increased significantly in the E0.05+DHT0.015 mg/kg group, E0.05+DHT0.05 mg/kg group, E0.05+DHT0.15 mg/kg group, E0.05+DHT0.5 mg/kg group, and E0.05+DHT1.5 mg/kg group (P<0.05). In addition, when the exogenous DHT concentration was constant, the MVD increased with the increase of exogenous E concentration in all the groups; Among them, compared with the control and castration group, the MVD increased significantly in the DHT0.15+E0.015 mg/kg group, DHT0.15+E0.15 mg/kg group, and DHT0.15+E0.5 mg/kg group (P<0.05).Conclusions: Androgens carried an important role in the regulation of prostatic MVD in SD rats, and the decrease of DHT concentration can induce a decrease in prostatic MVD. In contrast, prostatic MVD can be increased with the increase of DHT concentration. In addition, prostatic MVD can be increased gradually with the increase of estrogen concentration.

Berberine ameliorates testosterone-induced benign prostate hyperplasia in rats

Introduction Benign prostatic hyperplasia (BPH) is a major urologic problem that mostly develops in older males. Oxidative stress and inflammation influence the occurrence of BPH. Berberine (BBR) is a natural ingredient that has antioxidant and anti-inflammatory properties. The current research aims at examining the effects of BBR on testosterone-stimulated BPH in rats. Methods Animals were randomly categorized to six groups. In the control group, normal saline and olive oil were injected as the vehicle. BPH group: received testosterone (3 mg/kg, subcutaneous, 28 days), BPH + BBR groups; received BBR (25 and 50 mg/kg, p.o, 28 days), BPH + finasteride groups: received finasteride (1 mg/kg, p.o, 28 days), BBR (50 mg/kg, p.o, alone) was administered for subjects in the BBR group. On the 29th day, after anesthesia, cervical dislocation was used to kill the subjects. Serum concentration of testosterone and dihydrotestosterone was measured and prostate tissues were excised and used for biochemical, inflammation, and histological analysis. Results BBR prevented increased serum concentrations of testosterone and dihydrotestosterone. BBR considerably reduced BPH-stimulated oxidative stress and inflammation through preventing the rise in lipid peroxidation and nitrite concentration and declined the accumulations of pro-inflammatory cytokines (e.g. interleukin 1β and tumor necrosis factor α) and declining the depletion rate of GSH and the function of catalase and superoxide dismutase. Histopathological investigations reported that administration of BBR could suppress testosterone-stimulated BPH. Conclusion This study demonstrated that BBR could significantly prevent the development of BPH in rats.

Gingival Medicinal Signaling Cells Conditioned Medium effect on the Osteoclast and Osteoblast number in Lipopolysaccharide-induced Calvaria Bone Resorption in Wistar Rats’ (Rattus novergicus)

The gingival medicinal signaling cells conditioned medium (GMSCs-CM) is a biocompatible material which possessed beneficial cytokine, anti-microbial peptide, growth factor that can be collected after culture. GMSCs- CM may inhibit bone resorption in order to improve the patient’s quality of life. In this study, the potential effect of GMSCs-CM on the number of osteoclasts and osteoblasts in Lipopolysaccharide (LPS)-induced calvaria bone resorption in wistar rats (Rattus novergicus) has been analyzed. Twenty-eight male and healthy wistar rats (R. novergicus) at the age of 1-2 months old with 250-300 grams body weight were divided into 4 groups, namely PBS group: 100μg PBS day 1-7; LPS group: 100μg LPS day 1-7; LPS and GMSCs group: 100μg LPS + 100μg GMSCS-CM day 1 1-7, GMSCs group: 100μg M-GMSCs day 1-7. Escherichia Coli LPS was used to induce the bone resorption on the calvaria with subcutaneous injection. GMSCs-CM was collected after passage 4-5 then injected subcutaneously on the calvaria. All samples were examined on the. 8th day through cervical dislocation. The number of osteoblasts and osteoclasts in calvaria was then observed under 400x magnification. One Way ANOVA and Tukey HSD were conducted to analyze differences between groups (p<0.01). The number of osteoclasts in calvaria decreased significantly in the LPS + GMSCs-CM group compared to LPS group (p<0.01). The number of osteoblasts in calvaria increased significantly in the LPS + GMSCs-CM group compared to LPS group (p<0.01). GMSCs-CM can reduce the amount of osteoclast significantly and increases the production of osteoblast in LPS-induced calvaria bone resorption in wistar rats (R. novergicus).

EXTRA VIRGIN OLIVE OIL EFFECT IN THE COMBINATION OF ALCOHOL AND BENZENE ON THE KIDNEY OF RATTUS NOVERGICUS

Objective: This study investigated the use of extra virgin olive oil (EVOO), which is said to possess anti-inflammatory components as an ameliorative substance for the induced inflammation. Methods: Forty-eight female Wistar rats were used for this study. They were divided into eight (1–8) groups (n=6); Control, Ethanol, Benzene, E+B, E+O, B+O, E+B+O with the administration of 25% Ethanol, 200 mg/kg/b.w Benzene, 2 ml (O) EVOO in respect to the tagged grouping names. Animals were euthanized through cervical dislocation after the last day of administration and the liver was excised and part was fixed in formalin solution of 10% for histological processing and the other part was homogenized for biochemical assay in phosphate buffer before centrifugation. Results: Histological evidence creates an understanding of the nephrons in which benzene and ethanol manifest likewise the mitigated ability of EVOO. The relative organ weight provides information on the extent of the damage on nephrons. The kidney test level is significantly expressed. Conclusion: Ethanol and benzene in combination cause serious damage; also, they separately induce nephrons dysfunction. EVOO was shown to mitigate these nephrons damages.

THE EFFECT OF CONSUMING AVOCADO (Persea americana) ON MICE (Mus musculus) SPERM QUALITY

The nutrients such as protein and vitamin are proven to improve the sperm quality. One nutrient rich fruit is avocado round green variety, which contains vitamins A, C and E higher than other varieties. This study aimed to determine the effect of consuming avocado on the mice sperm quality. This experimental study was using a Completely Randomized Design with four treatments, each with eight replications. Avocado dosage treatments were control, 75 % w/v, 100% w/v, and 133% w/v , with the number of mice were 32 age four weeks. Avocado was given three times a day, each 0.5 mL for six weeks besides the main food. The observed response was the sperm quality including number, motility and morphology. Sperm was taken from the epididymis after the mice were killed by cervical dislocation method. Sperm was made a suspension using 0.9% NaCl solution and a smear preparation to observe the sperm morphology while staining with basic stain crystal violet. The sperm quality was observed by Neubauer's counting rooms through a multimedia microscope. Data was analyzed using Kurskal Wallis test for the sperm number and Anova test for motility and morphological responses. The result showed that avocado had a significant influence for reproductive health, particularly for increasing spermatozoa quality, including concentration, motility and morphology of sperm. The higher dose of avocado given the higher quantity and quality of sperm resulted.

The effect of Dichlorvous (Sniper) Inhalation on the Histology of the Lungs of Adult Male Wistar Rats

The organophosphate, dichlorvous (Otapia-pia) formulated in varying concentrations as insecticides is utilized by several individuals in most remote places of Nigeria due to its affordable value and accessibility. However, this present study is conducted to investigate the adverse effect of the exposure of this substance on the respiratory system (lungs) of male albino wistar rats. Twenty (20) albino wistar rats comprising of all males weighing between 150-230g were divided into four groups (A-D) of five animals each. Group A received only water, feed, and served as the control. Group B was exposed to dichlorvous inhalation 3hrs daily for a period of two weeks; Group C was exposed to dichlorvous inhalation 6hrs daily for a period of two weeks; while Group D was exposed to dichlorvous inhalation 10hrs daily for a period of two weeks. Twenty four hours after the last exposure, the animals were sacrificed by cervical dislocation and dissected. The lungs were weighed and fixed in 10% formal saline for histological studies. The body weight of the experimental groups decreased insignificantly when compared with the control group. The lungs weight increased significantly when compared to the control groups. Histological observation revealed a moderate to severe effects on the lungs with severe consolidated inflammatory exudates (discharge of fluids from pores), a moderate intra-alveoli hemorrhage, and a severe dilation of alveolar septa which is evident as emphysematous changes. This result revealed that the exposure to dichlorvous causes distortion of the normal histological architecture of the lungs. Thus, showing that dichlorvous inhalation influences the normal physiologic mechanisms of the lungs.

Euthanasia Through Cervical Dislocation or CO2 Might Affect Keel Bone Fracture Prevalence in 30-Week-Old Laying Hens

Laying hens are susceptible to keel bone fractures due to continuous endogenous calcium resorption for eggshell formation. Although it is assumed that external trauma to the keel bone, e.g., due to collisions, is the main cause for fractures, accumulated forces or asymmetric load on a weakened bone might contribute to the high keel bone fracture prevalence found in commercial laying hens. The objective of this study was to investigate whether forces applied to the keel due to involuntary convulsions and uncontrolled wing flapping during euthanasia have the potential to cause keel bone fractures. Two hundred and seventy Dekalb White laying hens were euthanized at 30 weeks of age using cervical dislocation (n = 60) or CO2 (n = 210). All hens were radiographed immediately before and after euthanasia. Radiographs were compared side by side to detect new fractures. Four out of the 270 hens (1.5%) obtained a fracture during euthanasia. Specifically, 0.95% of hens euthanized with CO2 (2 out of 210) and 3.3% of hens euthanized through cervical dislocation (2 out of 60) obtained a euthanasia-induced fracture. All four hens with a euthanasia-induced fracture had signs of damage to the keel before euthanasia, indicating that pre-existing fractures could affect fracture susceptibility. Based on our results, we cannot rule out that convulsions during euthanasia can cause keel bone fractures in laying hens. In studies investigating keel bone integrity in birds euthanized with CO2 or cervical dislocation, fracture prevalence might be overestimated. Future research is needed to assess whether euthanasia might be more likely to cause keel bone fractures in older birds and to quantify the frequency and strength of convulsions as a potential cause of fractures.

Leaf ethanolic extract of Etlingera hemesphaerica Blume mitigates defects in fetal anatomy and endochondral ossification due to mercuric chloride during the post-implantation period in Mus musculus

This study aimed to investigate the effectiveness of leaf ethanolic extract of Etlingera hemisphaerica (LE3H) in reducing defects in fetal anatomy and endochondral ossification in mice induced by HgCl2 during the post-implantation period. Pregnant mice were divided into four groups, each consisting of 10 dams, and received drink and food ad libitum. The first group was administered LE3H (E1), the second one HgCl2 (E2), the third one HgCl2+LE3H (E3), and the fourth was control (E0), administered double-distilled water only. HgCl2 (5 mg/kg bw) was administrated by injection intraperitoneally on gestation day (GD)9 and LE3H (0.39 mg/g bw) was administered by gavage on GD10. The treated and control animals were killed by cervical dislocation on GD18, dissected, and the morphologically normal living fetuses (MNLF) were collected. The MNLF of E0, E1, E2, and E3 from 5 dams were fixed with Bouin solution, and observed using the free hand razor blade technique for soft tissue examination. The remaining MNLF were fixed with 96% ethanol, and then stained with Alizarin Red S and Alcian Blue for ossification examination. Index of length of ossified part (ILOP) of humerus, index of width of ossified part (IWOP) of humerus, ILOP of femur, and IWOP of femur were calculated. E2 had higher cases of anatomical defects (74,6%) than E3 (48.9%), E1 (15.0%), and E0 (0%). E2 had humerus IWOP of 0.82±0.03, which was significantly lower than that of E0 (0.89±0.04) and E1 (0.89±0.03), while that of E1 and E0 was not significantly different from each other. Meanwhile, IWOP in E3 (0.88±0.03) was significantly higher than that in E2, but not different from that in E1 and E0. Thus, LE3H mitigated defects in fetal anatomy and endochondral ossification induced by HgCl2 in mice.