G2 Animals Research Articles

Assessing Facial Thermal Nociceptive Response in Female Dogs After Elective Ovariohysterectomy Anesthetized with Isoflurane and Treated with Cannabidiol and Meloxicam Analgesia

Pain management requires the identification of certain indicators to recognize pain. Various tools have been suggested to achieve an objective evaluation, including infrared thermography (IRT). The objective of this study was to assess the facial thermal nociceptive response produced by the use of cannabidiol (CBD) alone and in combination with meloxicam in female dogs undergoing elective ovariohysterectomy anesthetized with isoflurane. Sixty-four female dogs of different breeds were randomly distributed into four study groups according to the treatment received. G1: Placebo group (n = 16); G2: Group receiving intravenous meloxicam as premedication (0.2 mg Kg−1) and every 24 h postoperatively 0.1 mg Kg−1 (n = 16); G3: Group treated with CBD (n = 16) at a dose of 2 mg kg−1 orally every 12 h; and G4: Group medicated with the combination of both treatments (n = 16). All treatments were administered for 48 h postoperatively. After the anesthetic surgical procedure, radiometric images were captured using IRT and physiological parameters during the events EBasal, E30min, E1h, E2h, E3h, E4h, E8h, E12h, E24h and E48h. Overall, it was found that the high, medium and low temperatures of the thermal windows of the eye, upper eyelid and lower eyelid, as well as the average temperature of the lacrimal gland in G1 between events, were significantly lower at E30min, E1h and E2h compared to EBasal (p = 0.01). Among treatments, a significantly higher temperature was observed in groups G2, G3 and G4 compared to G1 (p = 0.001) in the thermal windows of the upper eyelid, lower eyelid, lacrimal gland and ocular areas. Regarding physiological parameters, heart rate (HR) was higher in G1 compared to the animals in G2, G3 and G4 (p = 0.03). The respiratory rate (RR) was significantly lower in all four study groups during the postoperative events compared to their respective EBasal (p < 0.05), while among treatments, G2, G3 and G4 had a lower RR compared to G1 (p = 0.03). Mild hypothermia was observed in all study groups at E30min and E1h compared to EBasal (p = 0.001). No significant correlation was found between the temperatures of the assessed thermal regions and the physiological traits. In conclusion, CBD, whether administered alone or in combination with meloxicam, demonstrated comparable analgesic efficacy, which could control nociceptive cardiorespiratory and hemodynamic autonomic responses, as there were no significant changes in the facial thermal response between treatments G2, G3 and G4.

Enhanced Costimulation Blockade With αCD154, αCD2, and αCD28 to Promote Heart Allograft Tolerance in Nonhuman Primates.

Long-term renal allograft acceptance has been achieved in macaques using a transient mixed hematopoetic chimerism protocol, but similar regimens have proven unsuccessful in heart allograft recipients unless a kidney transplant was performed simultaneously. Here, we test whether a modified protocol based on targeting CD154, CD2, and CD28 is sufficient to prolong heart allograft acceptance or promote the expansion of regulatory T cells. Eight macaques underwent heterotopic allo-heart transplantation from major histocompatibility complex-mismatched donors. Induction treatment for donor bone marrow transplantation (BMT) was administered after a 4-mo delay period under TNX-1500 monotherapy. The BMT induction regimen comprised 1 (group 1, G1; n = 3) or 2 (group 2, G2; n = 5) doses of total body irradiation, thymic irradiation, and antithymocyte globulin, followed by 2 (G1) or 5 (G2) weekly doses of αCD2 and 5 weekly treatments with αCD28 and TNX-1500. During the delay period, 1 G1 graft was rejected and 2 (1 in each group) exhibited moderate rejection on protocol biopsy before BMT. Lymphocyte chimerism was seen in 3 of 5 G2 animals and in 1 of 2 G1 recipients. One G1 graft was rejected despite chimerism, whereas the other recipient succumbed to anti-cytomegalovirus treatment. Two G2 monkeys succumbed due to infection (cytomegalovirus, bacteremia) post-BMT and 3 due to posttransplantation lymphoproliferative disease. Intensive costimulation pathway blockade with αCD2, αCD154, and αCD28 promotes lymphocyte chimerism at the cost of high incidence of posttransplantation lymphoproliferative disease and opportunistic infections, preventing assessment of the effectiveness of the regimen to promote alloimmune tolerance.

Dietary zinc status is associated with ZnT3 (SLC30A3), IL-6 gene expressions and spinal cord tissue damage in spinal cord tissue in a cuprizone-induced rat Multiple Sclerosis model

The aim of this study was to investigate the effects of dietary zinc status on spinal cord tissue damage and ZnT3, IL-6 gene expressions in a cuprizone-induced rat Multiple Sclerosis (MS) model. The study was carried out on 46 adult male rats of the genus Wistar. The animals used in the study were divided into 5 groups (G) (Control 6, other groups 10). G1, Control. G2, Sham-MS: Carboxy-methyl-cellulose (KMS) solution in which Cuprizon was dissolved was given to rats by gavage daily for 8 weeks at the rate of 1 % of daily feed consumption. MS was formed by giving 1 % of the daily feed consumption cuprizon in KMS solution by gavage to the animals in G3, 4 and 5 for 8 weeks. G4 was fed with a zinc deficient (50 µg/kg zinc) diet. G5 was given intraperitoneal (ip) zinc sulfate (5 mg/kg/day) supplementation. MS formation in animals was determined by Rotarod tests and Myelin Basic Protein (MBP) gene expression analysis. ZNT3 and IL-6 gene expression levels in spinal cord tissue samples of animals by Real-Time-PCR method; MDA and GSH levels were determined by ELISA method. The highest spinal cord MDA and IL-6 levels were obtained in G3 and G4 (P<0.05). Zinc supplementation in G5 prevented the increase in the mentioned parameters and turned them into control values (P<0.05). The spinal cord GSH and ZnT3 levels of G3 and G4 were lower than all other groups (P<0.05). Zinc supplementation prevented suppression in the same parameters in G5 and reached the control values (P<0.05). The findings of the current study suggest that zinc supplementation in addition to treatment for MS may be beneficial in reducing the severity of the disease.

Calf immunization protocols with low-virulence isolates of Anaplasma marginale: Analysis of post-inoculation effects and protection against natural challenge

Bovine anaplasmosis is endemic and is of fundamental importance worldwide. Therefore, measures for controlling and preventing clinical diseases are warranted to ensure the reduction of associated economic losses. The objective of the present study was to assess the post-inoculation effects and protection conferred by three different protocols of inoculation of low-virulence live strains of Anaplasma marginale (UFMG1 and UFMG3) in field-challenged cattle. Sixty-eight Holstein calves with an average age of 17 days were randomly divided into four groups. The groups received two subcutaneous administrations spaced 40 days apart, at a dosage of 2 × 106 infected erythrocytes of the following A. marginale strains: G1 (UFMG1 + UFMG1); G2 (UFMG3 + UFMG3); G3 (UFMG1 + UFMG3); and G4 (control). Every two days, the animals were evaluated for rectal temperature, Packed Cell Volume (PCV), and blood smears. Blood samples were collected prior to inoculation, before the field challenge, and after the challenge period, nPCR and IFAT techniques were performed. There were no significant differences in rickettsemia levels, reduction in PCV, or antibody detection among the different inoculation strategies. Forty days after the second inoculation, 90 %, 84.6 %, and 90.9 % of the animals in G1, G2, and G3, respectively, tested positive using nPCR. After inoculation, the group G2, which received the UFMG3 inoculum, had a higher frequency of treatment (odds ratio of 6.7; 1.198-38.018 CI; p = 0.03), while groups G1 and G3 demonstrated similar treatment frequencies compared to the control. During the natural challenge phase, 13.3 % of animals in group G1 required treatment (odds ratio of 0.108; 0.018-0.635 CI; p = 0.014) compared to 58.8 % of the control group. Considering the results collectively, the protocol using the UFMG1 strain (G1) stands out for its potential to be safe and induce some degree of immunization against A. marginale, reducing the incidence of clinical disease and the need for treatment during natural challenge.

Metabolic and Reproductive Parameters in Young Female Rats Subjected to Feed Restriction and/or Ginger Oil Administration

Background It is well known that the medicinal plant ginger (Zingiber officinale) has anti-oxidative properties that shield tissues and organs against oxidative damage. Aims Forty-eight, 9-week-old Sprague Dawley female rats (159±13 g) were used to evaluate the effects of feed restriction and/or ginger oil supplementation on growth, reproductive parameters, and blood metabolites. Methods Animals were randomly divided into 4 treatment groups (12 rats/treatment) in 2 × 2 factorial arrangements: ad libitum feed (CON), 25% feed restriction (G2), ad libitum feeding with 0.2 mL/rat ginger oil (G3), and 25% feed restriction plus 0.2 mL/rat ginger oil (G4). Animals in groups G2 and G4 received 75% of their pre-treatment feed intake (25% restriction). Six rats from each group were sacrificed at the end of treatments for blood and tissue collection, while the remaining rats were mated with mature males. Results No ginger-by-feed restriction interactions were detected in any of the tested parameters. Body size and BW increased as the experiment advanced (P &lt; 0.01). However, G2 had the lowest BW towards the end of the treatment. Glucose was greater (P &lt; 0.05) in feed-restricted animals, while urea nitrogen was greater in animals on ad libitum feeding. Combined ovarian weights and reproductive tract weights were greater in ginger-supplemented than non-supplemented animals. Similarly, rats on ad libitum feeding had greater ovarian weights than those on restricted feeding. Conclusion Ginger oil supplementation appears to have favorable effects on reproductive tract development and ovarian weight regardless of feed restriction.

Utility of facial thermography in the recognition of acute pain in hospitalized cats (Felis catus)

Different clinical conditions have been associated with the presence of pain in animals, so understanding the pathophysiological changes associated with their response is essential in in-hospital clinical monitoring. The objective of this study was to describe the usefulness of facial thermography in the recognition of acute pain in hospitalized cats (Felis catus), for which 71 cats were studied with an average weight of 4.3±1.4 Kg and an age average of 4.4±3.5 years. Facial temperature values were obtained in different region of interest (ROI) and compared according to the values of the Grimace pain scale in cats according to the following subclassification: Group 1, control animals (n=10); Group 2, animals classified with score 0-3 (n=21); Group 3, animals classified with score 4-6 (n=30) and Group 4, animals classified with score 7-10 (n=10). The global average temperature of the facial (F), auricular (A), periocular (PO), nasal (N) and rectal (R) regions was 33.7±1.9; 31.4±3.6; 32.1±0.9; 28.9±4.5 and 38.3±0.8 oC respectively. A difference (p&lt;0.05) was observed between the temperature of A and PO as well as between PO and N, but not between A and N. The present study allowed us to show an increase (p=0.0484) in the temperature mean PO ROI in G4 animals compared to G1 and a correlation with R temperature and peak temperature in the PO region in all groups. It is concluded that the increase in temperature of the periocular region and the nasal plane observed through infrared thermal imaging of the facial surface of hospitalized cats suggests the presence of clinically relevant acute pain. Facial thermography is a safe and non-invasive diagnostic method that complements other methods of identifying pain in cats.

Treatment with Raloxifene Induces the Expression of Kisspeptin, Insulin, and Androgen Receptors in Bones of Castrated Adult Female Rats.

Objective To evaluate the effects of estrogen, raloxifene and genistein on the expression of KISS1 (kisspeptin), KISS1R (kisspeptin receptor), AR (androgen receptor) and INSR (insulin receptor) in the bones of ovariectomized rats. Methods Forty-eight adult rats were randomly divided into 6 groups, containing 8 animals each: G1-nonovariectomized control; G2-ovariectomized and treated with conjugated equine estrogens (50 µg/Kg/day); G3-ovariectomized and treated with raloxifene (0.75 mg/kg/day); G4-ovariectomized animal that received soy extract with genistein (300 mg/kg/day); G5-ovariectomized animal that received estrogen and genistein; and G6-ovariectomized animal that received estrogen and raloxifene. Three months after surgery, the castrated animals received the drugs orally daily for 120 days. All animals were sacrificed after this period, by deepening the anesthesia. The left tibia was removed for total RNA extraction and analysis of gene expression of KISS1 , KISS1R , AR and INSR , by quantitative real-time polymerase chain reaction (qRT-PCR). Results KISS1 was not detected in any of the treated groups. KISS1R , INSR and AR showed higher expression in the G3 group ( p < 0.001), while lower levels of transcripts for these genes were observed in G4 and G5. G2 animals showed hypoexpression of the evaluated genes. Conclusion The results indicate that raloxifene, alone or combined with estrogen, was able to induce the expression of genes associated with the recovery of bone tissue homeostasis in ovariectomized rats.

Metabolite profile of Passiflora mollisima leaf using UHPLC-ESI-MS and its potential anxiolytic activity in mice

Background: Traditionally, extracts of Passiflora mollisima leaves are used as anxiolytics, but the existing metabolites are unknown. Objectives: To identify by UHPLC-ESI-MS/MS some of the metabolites and the anxiolytic activity of the leaf extract. Materials and Methods: The extract was subject to UHPLC-ESI-MS/MS analysis and administered intraperitoneally (VIP) to 4 groups of mice: G1: white group 0.1mL of saline solution, G2: positive control group 1mg Kg¯¹ of diazepam and to treatment groups, namely, G3 50 mg Kg¯¹ and G4 100 mg Kg¯¹ and anxiety levels were evaluated with the light/dark transition test for mice. Results: Six flavone C-glycosides were tentatively identified, namely vicenin 2, lucenin 2, schaftoside, orientin, vitexin, and glucopyranosyl methyl luteolin. Lower levels of anxiety were observed in animals of groups G3 and G4 based on the number of transitions, with a mean of 11±2 and 21±2, respectively, compared to G1, which was 2±1 (p&lt;0.05). The groups displayed a significant difference among them (p&lt;0.05). There was an increase in the total time spent in darkness in G3 and G4 Passiflora-treated mice with a mean of 70±8 and 113±8 respectively, compared to G1, which was 8±1 with a p&lt;0.05. Conclusion: The metabolites identified have biological activities, neuroprotective, antioxidant, anti-inflammatory, antibacterial, and anticancer effects. The results of the effects of the 50mg Kg¯¹ and 100 mg Kg¯¹ doses of ethanolic extract of Passiflora mollisima leaves significantly decreased anxiety levels (p&lt;0.05). This information contributes towards its further use in a therapeutic, clinical setting.

A preliminary study on the dynamics of serum color in perspective to hemoglobin and bilirubin in indigenous sheep of Pakistan.

The present study is the first of its kind being reported for an indigenous sheep breed of Pakistan with objectives to (a) assess the diagnostic efficacy of a human-based "serum hemolysis reference palette" for sheep serum, (b) deduce normal reference intervals (RIs) for hemoglobin (Hb) and bilirubin, and (c) devise a novel serum color chart for on-field estimation of Hb and bilirubin through color matching of sheep serum. Apparently, healthy Sipli sheep (n = 130) were bled twice attaining whole blood and serum samples (n = 260). The study animals were grouped on the basis of gender,that is, males (n = 51) and females (n = 79) and age,that is, G1 (up till 1 year) (n = 41), G2 (from 1 to 2 years) (n = 46), and G3 (from 2 to 3 years) (n = 43). None of the 260 serum samples of the sheep matched the color given on the human-based "hemolysis reference palette." The G1 animals revealed marked variation in their serum color. Hence, on the basis of RIs, the serum samples (n = 178) of adult sheep (G2 and G3) showing three main color bands were used in devising a novel serum Hb and bilirubin estimation chart for adult sheep serum. In conclusion, the human-based serum hemolysis palette is not valid for sheep serum. The RIs attained in the study could provide a yardstick for assessment of health in indigenous sheep breeds whereas the serum color chart may be of value in estimating Hb and bilirubin in a quick, reliable, and cheaper way for the resource-poor settings of the world.

Hippocampal ZnT3 (SLC30A3) Levels Reflect Hippocampal Tissue Damage in Chronic Exercising Diabetic Rats

Objective: In this study, it was investigated how chronic exercise affects hippocampus tissue damage and ZnT3 levels in diabetic rats. Methods: The 40 adult rats wereused in the study were divided into 4 equal groups: Control (G1), Exercise Control (G2), Diabetes (G3), Diabetes+Exercise (G4). Diabetes was induced in animals in G3 and G4 by injecting intraperitoneal streptozotocin (STZ) twice, 24 hours apart. The animals in G2 and G4 were runedon the rat treadmill for 45 minutes daily for 4 weeks. MDA (spectrophotometric method) and ZnT3 (ELISA method) levels were determined in hippocampus tissue samples obtained from animals sacrificed at the end of the experimental procedures. Results: In the current study, the highest MDA and lowest ZnT3 levels in the hippocampus tissue were obtained in the diabetes group (G3) (P&lt;0.05). Chronic exercise prevented increased hippocampal tissue damage in diabetic rats and reversed decreased ZnT3 levels (P&lt;0.05). Conclusion: The results of our study showed that 4 weeks of chronic exercise could be prevent increased tissue damage in the hippocampus tissue of diabetic rats and ameliorate the decreased ZnT3 levels. The data obtained in this study indicate that ZnT3 levels in diabetic rats may be an indicator of hippocampal tissue damage.

Composition and hepatoprotective effect of geopropolis of Melipona subnitida

Background: Geopropolis produced by bees of the subfamily Meliponinae (Hymenoptera, Apidae) is used in Brazilian popular medicine for the treatment of different diseases. The aim of the present work was to evaluate the hepatoprotective potential of the hydroethanolic extract from geopropolis (HPE) produced by Melipona subnitida Ducke. Additionally, the chemical composition of HPE and antioxidant activity were also evaluated. Methods: The hepatoprotective potential of HPE was evaluated by serum biochemical profile and macroscopic and histological examination of the liver of 24 Wistar rats (Rattus norvegicus), distributed in four experimental groups: G1 (negative control), G2 (HPE), G3 (HPE plus acetaminophen), G4 (acetaminophen - positive control). The HPLC-DADESI-MS/MS method was used to evaluate the chemical composition of HPE and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method was used to measure the antioxidant activity of geopropolis. Results: While HPE alone (G2) caused no liver injury, the livers of G4 exhibited color and histological alterations, extensive hemorrhagic suffusions, friable consistence, yellowish-white multifocal areas, degeneration and diffuse hepatocyte necrosis, in addition to altered levels of urea and enzymes TGO (glutamic-oxaloacetic transaminase) and TGP (glutamic pyruvic transaminase). HPE was shown to exert hepatoprotective effect in G3 animals, only mild hemorrhage and focal areas of vacuolar degeneration and hepatocyte necrosis having been observed. The geopropolis analyzed contains a high diversity of phenolic compounds, including chalcones, flavones and flavonols. high antioxidant activity was determined (IC50=48 μg/mL). Conclusions: The hydroethanolic extract of the geopropolis produced by M. subnitida has a high diversity of phenolic compounds and a high antioxidant activity. It has hepatoprotective action, as evidenced by a reduction in the levels of enzymaraes and the severity of the macroscopic and histological changes in livers of Wistar rats subjected to acetaminophen induced liver changes.

Effect of Cow-Calf Supplementation on Gene Expression, Processes, and Pathways Related to Adipogenesis and Lipogenesis in Longissimus thoracis Muscle of F1 Angus × Nellore Cattle at Weaning

The aim of this study was to identify differentially expressed genes, biological processes, and metabolic pathways related to adipogenesis and lipogenesis in calves receiving different diets during the cow-calf phase. Forty-eight uncastrated F1 Angus × Nellore males were randomly assigned to two treatments from thirty days of age to weaning: no creep feeding (G1) or creep feeding (G2). The creep feed offered contained ground corn (44.8%), soybean meal (40.4%), and mineral core (14.8%), with 22% crude protein and 65% total digestible nutrients in dry matter. After weaning, the animals were feedlot finished for 180 days and fed a single diet containing 12.6% forage and 87.4% corn-based concentrate. Longissimus thoracis muscle samples were collected by biopsy at weaning for transcriptome analysis and at slaughter for the measurement of intramuscular fat content (IMF) and marbling score (MS). Animals of G2 had 17.2% and 14.0% higher IMF and MS, respectively (p < 0.05). We identified 947 differentially expressed genes (log2 fold change 0.5, FDR 5%); of these, 504 were upregulated and 443 were downregulated in G2. Part of the genes upregulated in G2 were related to PPAR signaling (PPARA, SLC27A1, FABP3, and DBI), unsaturated fatty acid synthesis (FADS1, FADS2, SCD, and SCD5), and fatty acid metabolism (FASN, FADS1, FADS2, SCD, and SCD5). Regarding biological processes, the genes upregulated in G2 were related to cholesterol biosynthesis (EBP, CYP51A1, DHCR24, and LSS), unsaturated fatty acid biosynthesis (FADS2, SCD, SCD5, and FADS1), and insulin sensitivity (INSIG1 and LPIN2). Cow-calf supplementation G2 positively affected energy metabolism and lipid biosynthesis, and thus favored the deposition of marbling fat during the postweaning period, which was shown here in an unprecedented way, by analyzing the transcriptome, genes, pathways, and enriched processes due to the use of creep feeding.

In vitro and in vivo anthelmintic efficacy of condensed tannins extracted from the seeds of alfalfa (Medicago sativa L.) against Haemonchus contortus infection

This study was designed to examine in vitro and in vivo anthelmintic efficacy of condensed tannins (CT) extracted from seeds of Medicago sativa on Haemonchus contortus in sheep. CT's in vitro anthelmintic effect was assessed at a 300 μg/ml concentration compared with albendazole (reference drug) at 10 μg/ml. The results showed that CT had a nematocidal effect on H. contortus, and the cuticle of the adult worm appeared to be its initial target. For the in vivo experiment, nine 3-month-old helminths-free lambs were distributed into three groups. Group 1 (n=3) was challenged only as infected untreated controls; Group 2 (n=3) was treated with condensed tannin, and Group 3 (n=3) was treated with albendazole. Fecal and blood samples were collected every 3 days until the end of the experiment; for fecal egg count (FEC) and anti- H. contortus IgG titers determination, respectively. The lambs treated with the CT in G2 exhibited a pronounced decrease of mean FEC with great FECR% detected from the first-week post-treatment (PT) until the end of the experiment compared with G1 animals. The antibody levels gradually increased in G2 following the 2nd dose of CT treatment compared to other groups. A brilliant consistent relation between the elevation of IgG response and reduction of FEC was observed following the second booster dosing of the CT in G2. In conclusion, the CT evoked strongly in vitro and in vivo anthelmintic activity against H. contortus and could be used as a natural alternative treatment of high potency against haemonchosis in sheep.

Serum concentrations of folic acid and cobalamin and energy metabolism of ewes as a function of the energy density of the diet, peripartum period, and pregnancy toxemia.

The aim of this study was to determine the serum levels of folic acid, cobalamin, and indicators of energy metabolism during the peripartum period of ewes submitted to different diets and with or without pregnancy toxemia. Forty Santa Inês ewes were divided into three groups, a group of 20 animals fed only roughage-diet with low energy density (G1), a group of 10 ewes submitted to diet containing roughage and concentrate-diet with high energy density (G2), and a group of 10 ewes with diagnosis of pregnancy toxemia (PT) (G3). In the ewes of groups G1 and G2, blood samples were collected -30, -15, -7days pre-partum, on the day of partum, and 7, 15, and 30days postpartum; blood samples from G3 animals were collected during the veterinary clinic care. Significant variations were observed in the plasma concentration of glucose and serum folic acid, cobalamin, fructosamine, NEFA, and β-hydroxybutyrate in the groups fed diets with different energy levels, with and without PT (P < 0.05). A positive correlation of folic acid and cobalamin was observed with plasma glucose and serum NEFA concentrations, in addition to negative correlation of glucose with NEFA and β-hydroxybutyrate and positive correlation between the latter biomarkers of energy metabolism. The ewes diagnosed with PT showed higher levels of folic acid, NEFA, and β-hydroxybutyrate (P < 0.05). Ewes fed diet containing roughage and concentrate present higher serum levels of folic acid, cobalamin, and glucose, in the pre-partum period and in the lactation period. The results obtained contribute to better understanding of the application of blood concentration tests for folic acid and cobalamin in the energy metabolism and in PT in ewes.

Effect of Methotrexate and Matricaria Chamomile on testis tissue and some antioxidants in Albino rats

Objective: Depending on the antioxidant as well as anti-inflammatory properties of Matricaria Chamomile (MC), the current study aimed to investigate if MC could protect testicles against methotrexate (MTX)-induced testicular damage. Methods: 24 male Wistar albino rats were randomly separated into four groups (n=6/each) as follows. G1, the control group was given a single oral daily dose of distilled water (D.W) for 30 days; G2, (MTX group) rats were administered a single oral daily dose of D.W. and in day 28 the animals have received a single IP dosage of MTX; G3 six rats were given an aqueous extract of chamomile flowers at a dose of 200 mg/kg dissolved in 1 ml D.W. and on day 28 the rats have received a single intraperitoneal dose of MTX, and the last group was six rats received a methanolic extract of chamomile flowers at a dose of 200 mg/kg dissolved in 1 ml D.W. and on the day 28 the animals were received a one dosage intraperitoneal of MTX. At the end of the experiment, all animals were anesthetized and sacrificed, blood was also collected for the biochemical test as well as the testicular tissues were removed for histology examination. Results: According to biochemical assessments, notably there was no significant difference between the MTX and the other studied groups. In terms of histological examinations of animals treated with MTX, several pathological changes including vacuolization of seminiferous tubules with a reduced population of germinal epithelium were seen as compared to the control. Animals of G3 and G4 (pre-treatment with equates and alcoholic extract of MC before MTX administration) also showed some notable histopathological changes including disorganization of seminiferous tubules and tubular vacuolation. Conclusion: When comparing the control group, examination of testes with H&amp;E staining showed that pre-treatment with Matricaria Chamomile did not affect MTX-induced testicular damage.

Dehydrated Human Amnion/Chorion Membrane Allografts as an Adjunct Wound Healing Therapy in Diabetic Rats.

Chronic, non-healing wounds pose a serious public health issue and the need for new treatment methods is paramount. Dehydrated human amnion/chorion membrane has potential wound healing properties, due to the enrichment of growth factors and anti-inflammatory properties. However, its auxiliary advantage on diabetic wounds with demonstrated safety and efficacy in animal models has not been extensively documented. This study aimed at evincing the wound-healing property of dehydrated human amnion chorion membrane in diabetic and non-diabetic rats. An excisional wound model was developed in 36 male Sprague-Dawley rats that were randomly classified into six groups for two experiments. The non-diabetic rat group included non-diabetic control (G1), dHACM treatment (G2), and dHACM dressing + saline-treatment (G3); (n = 6). Similarly, the diabetic group included diabetic control (G4), dHACM treatment (G5), and dHACM dressing + saline-treatment (G6); (n = 6). The results of wound contractility rate, re-epithelialization, grading of granulation tissue, and collagen deposition from histopathological observation demonstrated that in comparison with the other groups (G1, G2, G4, and G5), the animal groups treated with dHACM dressing + saline-treatment (G3 and G6) had superior regenerative effects in excisional wound model. Also, in the animals of G5 and G6 of the diabetic group, there was no statistically significant difference (P > 0.05) in the levels of glucose, urea, creatinine, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphate (ALP), when compared to G4 animals during the experiment. It is evident from this study that dHACM could be applied as a potential wound healing biomaterial, especially in diabetic conditions.

Dietary Antimicrobial Peptides Improve Intestinal Function, Microbial Composition and Oxidative Stress Induced by Aeromonas hydrophila in Pengze Crucian Carp (Carassius auratus var. Pengze).

There is increasing evidence for the potential use of antimicrobial peptides as dietary supplements and antibiotic substitutes. In this study, we analyzed the differential effects of varying levels of antimicrobial peptides on the intestinal function and intestinal microbial and disease resistance of Pengze crucian carp. Approximately 630 experimental fishes were randomized in the control group (G0: 0 mg/kg) and in five groups supplemented with different doses of AMPs (G1: 100 mg/kg, G2: 200 mg/kg, G3: 400 mg/kg, G4: 800 mg/kg, and G5: 1600 mg/kg) and were fed for ten weeks. Three replicates per group of 35 fish were performed. The results showed that AMPs promoted intestinal villus development and increased intestinal muscular thickness (p < 0.05) and goblet cell abundance. The enzymatic activities of all groups supplemented with AMPs were effectively improved. AMP supplementation significantly enhanced the activities of antioxidant enzymes and digestive enzymes in the intestines of G3 animals (p < 0.05). Compared with G0 animals, AMP-supplemented animals regulated the expression of intestinal immune-related genes and exhibited significant differences in the G3 animal group (p < 0.05). The abundance of intestinal Firmicutes and Bacteroidetes increased in the AMP-supplemented groups, but the Firmicutes/Bacteroidetes ratio was lower than that in the G0 group. AMP supplementation also decreased the abundance of Fusobacterium while increasing the proportion of Actinobacteria (p < 0.05). After Aeromonas hydrophila infection, the expression levels of anti-inflammatory factors in the intestinal tract of G3 animals were significantly upregulated, and the level of the proinflammatory factor was decreased (p < 0.05). The intestinal Cetobacterium levels of G3 animals were significantly increased (p < 0.01), while the Proteobacteria levels were decreased, and the intestinal goblet cell proliferation was significantly lower than that of G0 animals (p < 0.05). This indicates that groups supplemented with AMPs have better disease resistance than the G0 group and can rapidly reduce the adverse effects caused by inflammatory response. Taken together, the present results suggest that AMP supplementation can improve intestinal function and intestinal microbial and pathogen resistance in Pengze crucian carp.

Induced arthritis, hepatocytes cytotoxicity and non-steroidal anti-inflammatory drugs (NSAIDs) – experimental study

This study aimed to histologically evaluate the pharmacological activity of meloxicam, COX-2 inhibitor, on liver function in arthritic rats. Meloxicam is a non-steroidal anti-inflammatory drug (NSAID), COX-2 inhibitor, generally prescribed in the area of rheumatology. Thus, 18 Wistar rats were divided into 3 groups with 6 animals each: G1 (Negative Control), G2 (Arthritis induced with Zymosan [Zy]), G3 (Arthritis induced with Zymosan [Zy] and treated with meloxicam). The G3 treatment started on the 21st day after induction. The animals were sacrificed, by overdose of Ketamine/Xylazine, seven, 14 and 21 days after the beginning of the meloxicam treatment. Liver samples were collected from the animals, fixed in Millonig buffer containing 10% formaldehyde and processed for conventional histological analysis. G1 animals showed absence of inflammation and degeneration of hepatocytes. In the G2 group, a slight cytoplasmic alteration was observed at seven and 14 days. At 21 days, intense cytoplasmic and nuclear disorganization was detected, accompanied by cell necrosis. In G3, large spacing between sinusoid capillaries was detected. In some 14-day-old animals, amyloidosis was observed, with the presence of a large number of vacuolated hepatocytes involved in an amorphous mass of necrotic cells. Most of the hepatocyte nuclei had compacted chromatin, evidencing cellular degeneration. Meloxican, used as an anti-inflammatory in the synovial region, showed that, in addition to its beneficial effects for the treatment of rheumatological diseases, it is capable of, when administered orally, promoting a high level of liver damage.

Ameliorate role of alpha lipoic acid on the beta amyloid accumulation and some serum oxidant / antioxidants levels in brain aging induced by d-gal in male rats

Alpha lipoic acid has both hydrophilic and hydrophobic properties and is widely dispersed in cellular membranes and the cytoplasm. It is one of the most powerful antioxidants for cells. The current research looked at the possible therapeutic benefits of alpha lipoic acid) in a male rat model of D-galactose-induced brain aging. G1 normal as control group, G2 animals were given 200 mg/kg bw.IP. daily of D-gal group for 30 days, and G3 animals were given 100 mg/kg. G4 D-gal + alpha lipoic acid group200mg/kg bw.IP. daily with 100 mg/kg. IP every day for 30 days.The accumulation of Beta amyloid in the brain and changes in serum oxidant/antioxidant levels Increased levels of oxidative stress indicators our data demonstrate that the D-gal aging model causes an increase in brain Beta amyloid deposition, as well as a substantial rise in serum MDA, GSH, CAT, and SOD levels. To summarize, our research focused on the brain damage caused by D-gal, which can be mitigated by intubating 100 mg/kg B.W.IP. of ALA to counteract its negative effects.

Effect of Caffeine and Vitamin D3 on the proliferative and hypertrophy zones of epiphyseal cartilage of neonate mice Femur

Objective: To evaluate the effect of caffeine and vitamin D3 on the height of proliferative and hypertrophy zones of epiphyseal cartilage of developing femur of BALB/c mice.&#x0D; Study design: Laboratory based randomized control trial.&#x0D; Place and duration of study: The study was conducted at Anatomy Department, Army Medical College (AMC), Rawalpindi, in collaboration with National Institute of Health (NIH), Islamabad from October 2014 to October 2015 (1 year).&#x0D; Material and Methods: A total of thirty (30) pregnant BALB/c mice, weighing 26-28 g, were taken and divided into three equal groups of 10 mice each. The control group G4 was given normal diet with water ad libitum. In addition to the same diet, animals in experimental group G5 were given 10 mg of caffeine per 100 g body weight once a day on alternate days through oral gavage for 21 days. All animals of group G6 for 21 days were given caffeine 10mg/100gm body weight on alternate day and vitamin D3 0.1µg per day by oral gavage. At completion of the experiment, effect of caffeine and vitamin D3 on the height of proliferative and hypertrophy zones of epiphyseal cartilage of neonate mice's femur was analysed. &#x0D; Results: The mean ±SD of height of proliferative and hypertrophy zones of control group G4 animals was found to be 540±10.99μm and 164±6.609μm, respectively. The mean ±SD of height of proliferative and hypertrophy zones of experimental group G5 animals was observed as 443.5±12.258μm and 138.25±6.129μm, respectively. The mean ± SD of height of proliferative and hypertrophy zones of experimental group G6 animals was measured as 474±3.8388μm and 144.25±3.7257μm, respectively.&#x0D; Conclusion: Caffeine intake modified the height of proliferative and hypertrophy zones of the epiphyseal cartilage of developing femur; however treatment with vitamin D3 mitigated this effect.&#x0D; Key words: Caffeine, Femur, Hypertrophy zone, Proliferative zone, Vitamin D3.