Tail Blood Research Articles

The differences of parasitemia and lungs size in malaria-associated acute respiratory distress syndrome (MA-ARDS) and non-MA-ARDS in mice infected with Plasmodium berghei ANKA

Malaria-associated acute respiratory distress syndrome (MA-ARDS) is characterized by extensive infiltration of leukocytes, microhemorrhages, vasogenic edema, changes in lung color, and a significant increase in the weight of the lung. This study was aimed to find out the differences in parasitemia and lung size in MA-ARDSand non-MA-ARDSin mice infected with Plasmodium berghei ANKA. Sixteen male BALB/c mice were infected with P. berghei ANKA, and daily parasitemia was observed on Giemsa-stained tail blood smears. Mice were sacrificed when parasitemia reached ±20%. Simultaneously eight uninfected mice were used as negative control (NEG). The statistical analysis was done using Kruskal Wallis, Mann Whitney U tests, and Spearman correlation test. The results showed that there were significant differences in parasitemia (p=0.001), weight (p=0.001), and lung length (p=0.021) between the MA-ARDS and non-MA-ARDS groups. Comparison of NEG and MA-ARDS resulted in a significant difference in lung size (p=0.05). When non-MA-ARDScompared with NEG groups, it showed a significant difference in lung width (p=0.001). However, there was no significant difference in lung weight and length (p>0.05). Spearman correlation test showed that there was a strong correlation between parasitemia with weight (p=0.000), length (p=0.001), and lung width (p=0.017). The findings indicated that parasitemia played a role in the development of MA-ARDS in mice infected with P. berghei ANKA and influenced the size of the lung.

Antimalarial activity of Virgin Coconut Oil against Plasmodium berghei ANKA in mice

Malaria remains a health problem in various parts of the world. A decrease in antioxidant levels during malaria infection cause oxidative stress, therefore exogenous antioxidant is needed. Virgin Coconut Oil (VCO) can be a source of exogenous antioxidants due to the antioxidant activity of VCO, which was contributed majority by phenolic acids content. Antioxidants play a role in countering the effects of free radicals by inhibiting fat peroxidation that cause erythrocyte cell wall stronger and not easily ruptured. Pharmacological properties of VCO including anti-inflammatory, anti-oxidant, antiviral, and antiprotozoal properties have been reported. Therefore, this study aimed to find out the antimalarial activity of VCO by evaluating the parasitemia and percentage of inhibition to the growth of parasite. Thirty male BALB/c mice were infected intraperitonially with 1×106 Plasmodium berghei ANKA-infected erythrocytes. The mice were than randomly divided into five groups: positive control (PC) group was given 187.2m g/kg BW of dihydroartemisinin phosphate, negative control (NC) group was only given sterile water; G1, G2, and G3 groups were given 1 ml, 5 ml, and 10 ml/kg BW of VCO, respectively. VCO treatments were given for 4 consecutive days started from 24 hours post infection. Parasitemia was determined daily on Giemsa-stained tail blood smear, further the percentage inhibition was calculated. The results showed that parasitemia in VCO-treated mice were lower than that of NC group, but higher than that of PC group, indicated the antimalarial activity of VCO. The inhibition of VCO to the growth of parasite showed that G2 was higher (48.70%) than that of G1 (13.04%) and G3 (33.9%). The use of antioxidant therapy as a supportive therapy is one of alternatives in supporting the healing process of malaria patients and may reduce various risks that could potentially occur in patients.

Rapid Decrease and Subsequent Increase in the Serum Triglycerides Accompanied by <i>CD36</i> Transcript Increase in an Acute Stress Mice Model

In this article, we report the changes in serum triglyceride (TG) levels that occurred during repeated tail blood sampling using a mouse restrainer. We used three groups of mice, namely, “PBS-restrained” “PBS-unrestrained” and “mock-restrained”. The mice in the PBS-restrained and PBS-unrestrained groups were intraperitoneally (i.p.) injected with 100 mL PBS and tail blood sampling was performed at 1, 5, 8, 24, and 48 h after i.p. injection. For the mock-restrained group, no i.p. injection was performed whereas the subsequent tail blood sampling was similarly performed. During the tail blood sampling, the mice of the two “restrained” groups were placed inside the restrainer designed from an open-ended 50 mL conical tube. The blood from the mice in the PBS-unrestrained group mice was sampled from the tail held by the operator’s hands while being allowed to move on a stage. Strikingly, in all of the three groups, the serum TG level initially decreased to remarkably low levels (approximately 30 mg/dL) after several blood samplings were performed over 8 h. This decrease was followed by a 2 - 3-fold increase in the levels relative to that in the control mice in the subsequent 24 - 48 h time period. We concluded that the acute stress associated with blood sampling caused alterations in TG levels. Serum levels of free fatty acid showed only modest changes. Changes in TG levels were not associated with serum corticosterone levels but with a dramatic increase in CD36 transcript levels in the liver. The relevance of this finding to the previously reported release of lipoprotein lipase (LPL) from white fatty tissue into the plasma during acute stress is also discussed.

Contribution of Brain Processes to Tissue Loss After Spinal Cord Injury: Does a Pain-Induced Rise in Blood Pressure Fuel Hemorrhage?

Pain (nociceptive) input soon after spinal cord injury (SCI) expands the area of tissue loss (secondary injury) and impairs long-term recovery. Evidence suggests that nociceptive stimulation has this effect because it promotes acute hemorrhage. Disrupting communication with the brain blocks this effect. The current study examined whether rostral systems exacerbate tissue loss because pain input drives an increase in systolic blood pressure (BP) and flow that fuels blood infiltration. Rats received a moderate contusion injury to the lower thoracic (T12) spinal cord. Communication with rostral processes was disrupted by cutting the spinal cord 18 h later at T2. Noxious electrical stimulation (shock) applied to the tail (Experiment 1), or application of the irritant capsaicin to one hind paw (Experiment 2), increased hemorrhage at the site of injury. Shock, but not capsaicin, increased systolic BP and tail blood flow in sham-operated rats. Cutting communication with the brain blocked the shock-induced increase in systolic BP and tail blood flow. Experiment 3 examined the effect of artificially driving a rise in BP with norepinephrine (NE) in animals that received shock. Spinal transection attenuated hemorrhage in vehicle-treated rats. Treatment with NE drove a robust increase in BP and tail blood flow but did not increase the extent of hemorrhage. The results suggest pain input after SCI can engage rostral processes that fuel hemorrhage and drive sustained cardiovascular output. An increase in BP was not, however, necessary or sufficient to drive hemorrhage, implicating other brain-dependent processes.

Effects of Metformin and Sitagliptin on islet cell function in insulin-resistant prediabetic KKay mice

Objective: To investigate the effects of metformin and sitagliptin on the function of islet β cells in insulin resistance pre-diabetic KKay mice. Methods: Thirty 6-week-old KKAy mice were randomly divided into two groups: normal diet fed group (NC group, n=10) and high-fat diet fed group (n=20). At 8 weeks, KKAy mice were randomly divided into two groups: metformin intervention group (met group, n=10) and sitagliptin intervention group (SP group, n=10), which were treated by gavage for 8 weeks. Glucose tolerance was measured by oral glucose tolerance test (OGTT), serum insulin level and plasma lipid level were measured by tail blood sampling, and HOMA-β and HOMA-IR were calculated. The mice were killed after blood collection, and the pancreas of KKAy mice was taken. The β cell volume of each group was compared by immunofluorescence staining of insulin and glucagon, respectively. The proliferation and apoptosis of β cell were analyzed by Ki67/INS. The expressions of pancreatic transcription factors PDX-1 and MafA were detected by Western blot. Results: ① The OGTT results indicated that blood glucose of KKAy mice at fast, 30, 60 and 120 min after oral administration of glucose in the Met and SP groups were decreased significantly compared with the NC group, and the area under the blood glucose time curve (AUC) was significantly reduced (P＜0.01, P＜0.01). There was no significant difference between the Met group and the SP group in blood glucose level at 30 and 60 min after oral administration of glucose. Compared with the SP group, the blood glucose of Met group at 120 min was decreased significantly (P＜0.05). There was no significant difference in AUC between the two groups. ② The results of the insulin tolerance test (ITT) indicated that, compared with NC, the fasting blood glucose and the blood glucose at 30, 60 and 90 min after insulin injection in KKAy mice in the Met and SP groups were decreased significantly, and the area under the ITT blood glucose curve (AUC) was increased significantly (P＜0.01), while there was no significant difference between the Met and SP groups. ③ In the NC group, the brightness of the areas of the islet β cells was low and the edges were scattered. After treated with metformin, the areas and brightness of the β cells were increased. After treatment with sitagliptin, the area and brightness of the β cells were increased significantly. In the NC group, the α cells were disordered in the islet distribution and the brightness was large. After the administration of metformin, the α cell area and the brightness were decreased, and distributed to the edge of the islet to a certain extent. After the administration of sitagliptin, there was a significant decrease in the area of the α cells, with a significant decrease in the brightness and distribution at the edge of the islet. ④ Compared with the NC group, the expression levels of pancreatic MafA in the Met group and SP group were increased significantly, which were 1.63 times and 1.58 times, respectively (P＜0.01, P＜0.01). There was no significant difference in the expression of pancreatic PDX-1 between the groups. Conclusion: In pre-diabetes mellitus KKAy mice with insulin resistance, metformin can maintain the function and morphology of pancreatic islets, and sitagliptin may promote the proliferation of islet βcells, improve the expression level of insulin transcription factor MafA, and prevent the occurrence and development of diabetes.

Effect of Angiotensin 1-7 on Myocardial Calcium Pump in Salt-Sensitive Hypertensive Rats

Objective — To investigate the effects of angiotensin 1-7 (Ang1-7) on plasma membrane ATPase isoform 1 (PMCA1) in salt-sensitive hypertensive rats. Methods — Thirty newborn male Wistar rats were selected to establish the salt-sensitive hypertensive rat model with sensory nerve injury, which were then randomly divided into 5 groups (n=5), including model group, Telmisartan group, Ramipril group, Ang1-7 group, and A-779 group. Another normal control group was established (n=5). After 4 weeks of intervention, the tail blood pressure of rats in each group was measured, and then the apical tissue of left ventricle was cut. The contents of AngⅡ and Ang1-7 in cardiomyocytes were detected by enzyme-linked immunosorbent assay. The expression of PMCA1 mRNA and protein in heart of salt-sensitive hypertensive rats were detected by RT-PCR and immunohistochemistry. Results — (1) Compared with the normal control group, the concentration of AngⅡ in the myocardium of salt-sensitive hypertensive rats increased (P < 0.05), which decreased after the intervention of Telmisartan and Ramipril (P < 0.05), and no change occurred after the intervention of Ang1-7 in concentration (P＞0.05). (2) Compared with the normal control group, the concentration of myocardial Ang1-7 in salt-sensitive hypertensive rats decreased (P < 0.05), and increased after the intervention of telmisartan and ramipril (P < 0.05), and increased after the intervention of A-779 (P < 0.05). (3) The expression of PMCA1 mRNA and protein in salt-sensitive hypertensive rats was increased compared with the normal control group (P < 0.05), and the expression of Ang-(1-7), telmisartan and ramipril was decreased compared with the model group (P < 0.05). The expression of p38MAPK mRNA and p-p38MAPK protein in the myocardium of salt-sensitive hypertensive rats was increased compared with that in the normal control group (P < 0.05), and the expression of Ang-(1-7), Telmisartan and Ramipril was decreased compared with that in the model group (P < 0.05). Conclusion — Ang-(1-7) may be involved in the regulation of cardiac calcium pump, inhibiting its overcompensation and delaying the occurrence of calcium pump inhibition in the early stage of salt-sensitive hypertension. Ang-(1-7) can inhibit the activity of p38MAPK and protect the heart, and its regulation on PMCA1 may be mediated by the expression of p38MAPK pathway.

Effect of Various Activation of the Skin TRPA1 Ion Channel on Cardiovascular Systemic Parameters in Hypertensive Rats

The effect of different activation of the skin TRPA1 ion channel on the parameters of the cardiovascular system was studied in rats with hereditary stress-induced hypertension while recovery from anesthesia. The activation of the TRPA1 ion channel was performed by application to skin its agonist - allyl isothiocyanate (AITC) in two concentrations 2.5% and 0.5%. The following parameters were measured that characterize the work of the cardiovascular system: Blood pressure (systolic, diastolic, and mean), heart rate, blood flow, and tail blood volume. Stimulation of the TRPA1 ion channel by 2.5% AITC resulted in a diminishing rate of increase in blood pressure and a sharp increase in heart rate while recovering from anesthesia.

Aerobic Exercise Increases Expression Of SUR1 And SUR2A Subunits Of K ATP -channels In Murine Diabetic Ventricular Cardiomyocytes.

Diabetes mellitus (DM) is one of the most prevalent and rapidly increasing non-communicable chronic diseases around the world. It has been related to left ventricular disfunction, as well to its electric remodeling. Cellular and molecular underlying mechanisms remains unveiled, at least in part. Phenotypic consequences includes QT Interval prolongation, ventricular arrhytmia, and cardiac sudden death. KATP-channels are considered as couplers of metabolism to membrane electric behavior in ventricular cells. At early DM stages, reduction in KATP-channels expression has been claimed as one of the main cardiac dysfunction underlying mechanisms. Physical exercise (PE) is postulated as a preventive and therapeutic intervention for diabetic cardiomyopathy. PURPOSE: to explore aerobic PE effects on expression of KATP-channel sulphonylurea receptors (SURs) in diabetic cardiomyocites. METHODS: 3 weeks male Wistar rats were randomly asigned to four groups, control with (EC) and without exercise (NEC); diabetes with (ED) and without exercise (NED) groups (n = 5, each). IP streptozotocin at 100 mg/Kg dosis was used to induce DM. DM was considered when glycaemia was ≥250 mg/dl in tail blood. EC and ED performed exercise in a treadmill 5 min at a 15 m/min pace, followed by 20 min at 20 m/min, along 13 consecutive days. NEC and NED spent similar periods of time in a nonfuctioning treadmill. At the end of the program animals were eutanized and tissue samples were obtained from the ventricular apex. Subsecuently, Real Time Polymerase Chain Reaction (rtPCR) was used to SURs expression quantitation. RESULTS: significant differences were found for weight and glycaemia between NEC and EC when compared to NED as well as to ED (p = 0.05). Relative expression (mRNA SURx/β-actin genes) was higher in ED: for SUR1, ED vs. NED were 10.54 ± 3.53 vs 1.32 ± 0.38 (p = 0.007), as well when compared to NEC (1.29 ± 0.47, p = 0.01). Similarly, SUR2A expression was higher in ED than in NED; 10.68 ± 2.54/1.22 ± 0.45 (p = 0.002), and than in NEC (1.18 ± 0.33, p = 0.06). No differences were found in the expression of SUR2B. CONCLUSIONS: Aerobic PE increases SUR1 and SUR2A isoforms of the KATP-channels sulfonylurea receptors. This phenomenon could be related to cardioprotective effects of aerobic physical exercise in Diabetes Mellitus.

Roles of Averrhoa bilimbi Extract in Increasing Serum Nitric Oxide Concentration and Vascular Dilatation of Ethanol-Induced Hypertensive Rats.

The considerably high incidence of cardiovascular disease in Indonesia has attracted scientists to investigate various plant and fruit extracts as preventive agents. Averrhoa bilimbi (AB) is rich in bioactive constituents that may be effective in preventing indicators of hypertension. This study evaluated the roles of AB extract in increasing serum nitric oxide (NO) concentration and vascular dilatation in ethanol-induced hypertensive rats. A total of 24 male Wistar rats (Rattus norvegicus) were divided equally into 4 treatment groups (n=6): P0 (control group, administered placebo); P1 [administered captopril 3 mg/kg body weight (BW) orally]; P2 (administered AB extract at 20 g/kg BW); and P3 (administered AB extract at 40 g/kg BW). The AB extract was obtained from fresh AB macerated in 96% ethanol and was subjected to bioactive compounds identification using thin layer chromatography. After pretreatment with ethanol for 15 days, treatments were administered daily for 14 days. All rats were measured for tail blood pressure by the tail-cuff method and NO concentrations by avidin-horseradish peroxidase sandwich-enzyme-linked immunosorbent assays. All rats were sacrificed to collect blood vessels for histopathology. The results showed that AB extracts contained flavonoids, saponins, polyphenols, essential oils, and anthraquinone. Treatment with AB extract at a dose of 40 mg/kg BW significantly increased NO concentrations (P<0.05). Histopathological analysis showed that AB extracts inhibited endothelial pyknosis, intimal body, and adventitial leukocyte infiltration of posterior vena cava blood vessels. These results suggest that the protective effect of AB extracts is associated with NO concentration in the blood by inhibiting blood vessel dysfunction.

Local vibration induced vascular pathological structural changes and abnormal levels of vascular damage indicators

BackgroundThe vascular component of the hand-arm-vibration syndrome (HAVS) is often characterized by vibration-induced white fingers (VWF). Active substances secreted by the vascular endothelial cells (VEC) maintain a dynamic balance but damage to the blood vessels may occur when the equilibrium is altered, thus forming an important pathological basis for VWF. This study was aimed at investigating vascular damage indicators as a basis for an early detection of disorders caused by vibration, using the rat tail model. Methods and resultsExperiments were conducted using a control group of rats not exposed to vibration while two exposed groups having different exposure durations of 7 and 14 days were randomly formed. Following exposure, the structural changes of tail tissue samples in anesthetized rats were observed. Enzyme-linked immunosorbent assay (ELISA) was used for analyzing four vascular damage indicators myosin regulatory light chain (MLC2), endothelin-1 (ET-1), vinculin (VCL) and 5-hydroxytryptamine (5-HT) in tail blood samples. We found that both vascular smooth muscle and endothelial cells displayed changes in morphology characterized by vacuolization and swelling in the vibration-exposed group. The levels of vascular damage indicators were altered under the vibration. ConclusionThe degree of vascular pathology increased with the longer duration exposure. Furthermore, the levels of MLC2, ET-1 and 5-HT in rat plasma were associated with vascular injury caused by local vibration.

Anorectic interaction and safety of 5-hydroxytryptophan/carbidopa plus phentermine or diethylpropion in rat.

Drug combinations are being studied as potential therapies to increase the efficacy or improve the safety profile of weight loss medications. This study was designed to determine the anorectic interaction and safety profile of 5-hydroxytryptophan (5-HTP)/carbidopa + diethylpropion and 5-HTP/carbidopa + phentermine combinations in rats. The anorectic effect of individual drugs or in combination was evaluated by the sweetened milk test. Isobologram and interaction index were employed to determine the anorectic interaction between 5-HTP/carbidopa and diethylpropion or phentermine. Plasma serotonin (5-HT) was measured by ELISA. Safety of repeated doses of both combinations in rats was evaluated using the tail sphygmomanometer, cardiac ultrasound, hematic biometry and blood chemistry. A single oral 5-HTP, diethylpropion or phentermine dose increased the anorectic effect, in a dose-dependent fashion, in 12 h-fasted rats. A dose of carbidopa at 30 mg/kg reduced the 5-HTP-induced plasmatic serotonin concentration and augmented the 5-HTP-induced anorectic effect. Isobologram and interaction index indicated a potentiation interaction between 5-HTP/30 mg/kg carbidopa + diethylpropion and 5-HTP/30 mg/kg carbidopa + phentermine. Chronic administration of experimental ED40 of 5-HTP/30 mg/kg carbidopa + phentermine, but not 5-HTP/30 mg/kg carbidopa + diethylpropion, increased the mitral valve leaflets area. Moreover, there were no other significant changes in cardiovascular, hematic or blood parameters. Both combinations induced around 20% body weight loss after 3 months of oral administration. Results suggest that 5-HTP/30 mg/kg carbidopa potentiates the anorectic effect of diethylpropion and phentermine with an acceptable safety profile, but further clinical studies are necessary to establish their therapeutic potential in the obesity treatment.

Effects of intraperitoneal and intracerebroventricular injection of cinnamaldehyde and yohimbine on blood glucose and serum insulin concentrations in ketamine-xylazine induced acute hyperglycemia.

This study was designed to investigate the effects of peripheral [intraperitoneal (IP)] and central [intracerebroventricular (ICV)] administration of cinnamaldehyde on concentrations of blood glucose and serum insulin in the acute hyperglycemia induced by ketamine/xylazine. Yohimbine (a α2-adrenoceptor antagonist) was used alone and in combination with cinnamaldehyde to explore the α2-adrenergic receptor contribution. A total of 48 rats were divided into eight groups with six rats in each for IP administration of normal saline, vehicle, cinnamaldehyde (25.00, 50.00 and 100 mg kg-1), yohimbine (0.50 and 2.00 mg kg-1) and cinnamaldehyde plus yohimbine. These rats were used again for ICV administration 15 days after the completion of IP experiment. During this 15 days period, the lateral ventricle of the brain was surgically cannulated for ICV administration of normal saline, vehicle, cinna-maldehyde (25.00, 50.00 and 100 µg per rat), yohimbine (5.00 and 20.00 µg per rat) and cinnamaldehyde plus yohimbine. Blood glucose levels were measured from tail blood using a glucometer and serum insulin concentrations were determined via enzyme-linked immune-sorbent assay kit. The increased levels of blood glucose and the decreased concentrations of serum insulin were significantly decreased and increased, respectively, by separate and combined IP and ICV administrations of cinnamaldehyde and yohimbine. The systemic effects of these chemical compounds were significantly greater than the central ones. Based on the results, it can be argued that cinnamaldehyde has a potential to induce anti-hyperglycemic and antihypoinsulinemic effects. Peripheral and central α2-adrenegic receptors might be involved in these effects of cinnamaldehyde.

Intradermal Immunization with Heat-Killed Klebsiella pneumoniae Leading to the Production of Protective Immunoglobulin G in BALB/c Mice.

Introduction:Klebsiella pneumoniae superbug is emerging as a serious health concern as resistance to last-resort antibiotics spreads. To bypass the therapeutic molecules used today, the development of an immunoprophylactic safe approach is of great clinical relevance. This study was conducted to determine the protective efficacy of antibodies elicited by killed vaccine against multidrug-resistant (MDR) K. pneumoniae.Materials and Methods:In this study, heat-killed MDR K. pneumoniae isolated from different clinical samples were employed for the intradermal immunization of 10 BALB/c mice. Two weeks after the third dose of immunization, the mice were intraperitoneally challenged with live K. pneumoniae and observed for 14 days. Tail blood was collected 7 days after each booster followed by cardiac puncture 14 days postchallenge. Bactericidal activity and antigen-binding capacity of the serum antibody produced by the vaccine were evaluated by serum bactericidal antibody (SBA) assay and ELISA, respectively.Results:In this study, 80% survival rates were observed at 14 days postchallenge among the immunized mice. Regarding SBA assay, 100% bactericidal activity of the immunized mouse sera was observed using 50% guinea pig complement at 1:10 serum dilution after 3 h of incubation, and all the pre- and postchallenge immunized serum immunoglobulin G antibody had significantly higher optical density values comparing the control mice in ELISA.Conclusion:In our study, intradermal immunization with heat-killed MDR K. pneumoniae produced protective antibodies in BALB/c mice. These findings suggest that the use of a first-generation vaccine provides the supply of a larger number of candidate antigens for eliciting required immune response.

Effect of 8-Week Exercise Program on Bone Biomarker Osteocalcin and Bone Histomorphometry Features in Male Rats

Background: The anabolic effect of physical exercise on osseous tissue is related to mechanical effort, leading to the osteogenic response by causing dynamic changes, which stimulate osteocytes through fluid shifts in their canalicular network. They produce signaling molecules that regulate bone formation and absorption by osteoblast and osteoclasts. Objective: To study the effect of 8-week exercise training programs on the histomorphometry of male rat femur bone including weight, length, thickness and the bone formation biomarker (osteocalcin). Methods: The study was done in the labs of College of Medicine, Al-Nahrain University, from September 2019 to February 2020. A thirty adult healthy male rat (albino rat), were selected and divided into three groups; the group (A) of rats with exercise training programs of treadmill running, for 8 weeks. The control group, (B) were kept under free movements without exercise. The group (C) was kept under restricted movements in small cages. Tail blood sample were obtained twice from all animals; at zero day and after 8 weeks, for measurement of osteocalcin. Then after 8 weeks all animals were sacrificed and dissected for extraction of femoral bone for measuring bone length, weight, thickness and bone ultrastructure under light microscope by staining with hematoxylin and eosin. Results: The osteocalcin, femoral bone weight, length, thickness, haversian thickness and lamellar thickness showed significant increase in value of group A in comparison to group B and group C which show a significant decrease in femoral bone thickness, haversian thickness, lamellar thickness, and osteocalcin level. Conclusion: Exercise training has an anabolic effect on bone, in contrast to restriction movement that cause catabolic effect on bone. Osteocalcin increases with exercise and could be used as a marker in monitoring the exercise program therapy. Keywords: Femoral bone weight, length, thickness, haversian thickness, lamellar thickness, osteocalcin Citation: Musleh AM, Hashim ZH, Jaafar HA. Effect of 8-week exercise program on bone biomarker osteocalcin and bone histomorphometry features in male rats. Iraqi JMS. 2020; 18(2): 145-154. doi: 10.22578/IJMS.18.2.9.

Effect of 8-Week Exercise Program on Bone Biomarker Osteocalcin and Bone Histomorphometry Features in Male Rats

Background: The anabolic effect of physical exercise on osseous tissue is related to mechanical effort, leading to the osteogenic response by causing dynamic changes, which stimulate osteocytes through fluid shifts in their canalicular network. They produce signaling molecules that regulate bone formation and absorption by osteoblast and osteoclasts. Objective: To study the effect of 8-week exercise training programs on the histomorphometry of male rat femur bone including weight, length, thickness and the bone formation biomarker (osteocalcin). Methods: The study was done in the labs of College of Medicine, Al-Nahrain University, from September 2019 to February 2020. A thirty adult healthy male rat (albino rat), were selected and divided into three groups; the group (A) of rats with exercise training programs of treadmill running, for 8 weeks. The control group, (B) were kept under free movements without exercise. The group (C) was kept under restricted movements in small cages. Tail blood sample were obtained twice from all animals; at zero day and after 8 weeks, for measurement of osteocalcin. Then after 8 weeks all animals were sacrificed and dissected for extraction of femoral bone for measuring bone length, weight, thickness and bone ultrastructure under light microscope by staining with hematoxylin and eosin. Results: The osteocalcin, femoral bone weight, length, thickness, haversian thickness and lamellar thickness showed significant increase in value of group A in comparison to group B and group C which show a significant decrease in femoral bone thickness, haversian thickness, lamellar thickness, and osteocalcin level. Conclusion: Exercise training has an anabolic effect on bone, in contrast to restriction movement that cause catabolic effect on bone. Osteocalcin increases with exercise and could be used as a marker in monitoring the exercise program therapy. Keywords: Femoral bone weight, length, thickness, haversian thickness, lamellar thickness, osteocalcin Citation: Musleh AM, Hashim ZH, Jaafar HA. Effect of 8-week exercise program on bone biomarker osteocalcin and bone histomorphometry features in male rats. Iraqi JMS. 2020; 18(2): 145-154. doi: 10.22578/IJMS.18.2.9.

Microsatellite Analysis of Genetic Diversity and Population Structure of the Iranian Kurdish Horse

Native breeds are essential for national stocks and genetic reservoir; therefore, the preservation of indigenous breeds is a key policy priority for countries around the world. Many conservationists would assert that genetic diversity is a prerequisite for adaptive evolution, and preserving genetic diversity will need conservation efforts for the long-term survival of domestic species. This study intended to evaluate the genetic diversity of the Iranian Kurdish horse population based on microsatellite indicators, which can partially prevent it from becoming extinct. Fifty-eight tail hair and blood samples were randomly collected from Kurdistan, Kermanshah, Ilam, West Azerbaijan, Isfahan, Kerman, Hamadan, and Tehran. Genomic DNA extraction was performed by a modified salting out method. The polymerase chain reaction amplification conditions were also separately undertaken for each marker. All microsatellite loci revealed polymorphisms in the studied population. Genetic variation was examined using 12 microsatellite loci (HMS7, HMS3, HMS2, HMS6, ASB2, ASB23, VHL20, HTG10, LEX33, ASB17, AHT4, and AHT5). We found that the means of the observed and effective number of alleles were 7.58 and 4.95, with the minimum and maximum values for each of these indices associated with the loci of HMS2 and ASB17, respectively. Moreover, the mean of observed and expected heterozygosity, polymorphism information content, and Shannon’s Information Index of the Iranian Kurdish population were 0.77, 0.78, 0.75, and 1.67, respectively, indicating a high degree of genetic diversity in the entire studied population. More specifically, we acquired a range of new alleles in the Iranian Kurdish horse breed that differed in their genetic structure to those of other Iranian breeds in other studies. This study provides an exciting opportunity to improve our knowledge of genetic information which will be beneficial as a base to identify purebred Kurdish horses for a further Iranian Kurdish horse genetic and breeding program.

Antimalarial activity of goat bile against Plasmodium berghei ANKA infection in BALB/c mice

Abstract Goat bile has been used by some Indonesian people to treat malaria and increase their stamina. This study aimed to prove whether goat bile toxic or not in BALB/c mice and to verify the antimalarial activity of goat bile at various concentrations in mice infected with Plasmodium berghei ANKA. Acute toxicity test was performed using twenty male BALB/c mice with an average body weight of 25 grams, which were divided into four groups. Mice were given 25%, 50%, and 100% goat bile, respectively, while negative control was given distilled water. Any change in weight, odor, color, agitation, appearance, color of urine and feces, coma, and death, were recorded. A different set of mice were infected with P. berghei ANKA. This study conducted using the posttest only control group design with four treatments and five replications. A four day-treatment of goat bile was given by oral gavage to find out its effect on parasitemia level. Infected mice were divided randomly into 4 groups, where the GBNeg group as negative control was given only distilled water. The GB25, GB50, and GB100 groups were treated with 25%, 50%, and 100% goat bile, respectively. The parasitemia was observed daily on Giemsa-stained tail blood smears of each mice. No death or other sign of toxicity was found in goat bile-treated mice. Goat bile showed anti-malarial activity. The parasitemia in all goat bile treated groups was lower compared with the negative control group. The ED50 of goat bile against the growth of parasite was 48,55 %. Goat bile is a potential source of new antimalarial therapies. Further investigations are recommended to yield new anti-malarial drug candidates. Keywords : Goat bile, parasitemia, Plasmodium berghei ANKA, ED50Correspondence : heny-a@fk.unair.ac.id

S‑1‑Propenylcysteine, a sulfur compound in aged garlic extract, alleviates cold‑induced reduction in peripheral blood flow in rat via activation of the AMPK/eNOS/NO pathway

Aged garlic extract (AGE) has been shown to improve peripheral circulatory disturbances in both clinical trials and experimental animal models. To investigate the effect of S-1-propenylcysteine (S1PC), a characteristic sulfur compound in AGE, on cold-induced reduction in tail blood flow of rat, Wistar rats were individually placed in a restraint cage and given the treatment with cold water (15˚C) after the oral administration of AGE or its constituents S1PC, S-allylcysteine (SAC) and S-allylmercaptocysteine (SAMC). After the cold-treatment the tail blood flow of rats was measured at the indicated times. The pretreatment with AGE (2 g/kg BW) and S1PC (6.5 mg/kg BW) significantly alleviated the reduction of rat tail blood flow induced by cold treatment. The effect of S1PC was dose-dependent and maximal at the dose of 6.5 mg/kg BW, whereas SAC and SAMC were ineffective. To gain insight into the mechanism of S1PC action, the concentration of nitrogen oxide metabolites (NOx) in the plasma and the levels of phosphorylated endothelial nitric oxide synthase (eNOS) and 5'-AMP-activated protein kinase (AMPK) in the aorta were measured. The pretreatment with S1PC significantly increased the plasma concentration of NOx as well as the level of phosphorylated form of AMPK and eNOS in the aorta after cold-treatment. The present findings suggest that S1PC is a major constituent responsible for the effect of AGE to alleviate the cold-induced reduction of peripheral blood flow in rat by acting on the AMPK/eNOS/NO pathway in the aorta.

Development of a cyclophosphamide stress test to predict resilience to aging in mice.

The concept of resilience, defined as the ability to recover from stress, is a potential platform to predict healthy aging. However, specific stress tests for resilience have not yet been fully established in humans so investigations in animal models are of interest. The chemotherapeutic drug cyclophosphamide (Cyp) was selected as a chemical stressor to investigate resilience response in C57Bl/6 male mice at 4, 15, and 28months of age. Following a single intraperitoneal injection of Cyp (100mg/kg), tail blood was collected for counting white blood cells (WBC) every other day for 25days, and physiological performance tests performed. Cyp induced a consistent pattern in neutrophil count in all three age groups, with a nadir at day 5 and a rebound at day 7 with different rates in each group. The neutrophil to lymphocyte ratio (NLR) showed an age-dependent rebound response 7days after Cyp injection, with a similar pattern of decline back toward baseline. Mice in the 15-month age group with high pre-injection Cyp NLR had significantly higher total WBC counts after Cyp injection compared with mice with low pre-injection Cyp NLR, indicating a correlation between NLR and Cyp-altered WBC counts. In addition, mice with high pre-injection Cyp NLR showed significant learning impairment compared with mice with low pre-injection Cyp NLR, suggesting low NRL intensity can predict resilience to age-related cognitive decline. These observations provide the rationale to translate findings from the mouse to humans in developing in vitro Cyp stress tests.

Transglutaminase 2 Has Metabolic and Vascular Regulatory Functions Revealed by In Vivo Activation of Alpha1-Adrenergic Receptor.

The multifunctional tissue transglutaminase has been demonstrated to act as α1-adrenergic receptor-coupled G protein with GTPase activity in several cell types. To explore further the pathophysiological significance of this function we investigated the in vivo effects of the α1-adrenergic receptor agonist phenylephrine comparing responses in wild type and TG2-/- mice. Injection of phenylephrine, but not a beta3-adrenergic agonist (CL-316,243), resulted in the long-term decline of the respiratory exchange ratio and lower lactate concentration in TG2-/- mice indicating they preferred to utilize fatty acids instead of glucose as fuels. Measurement of tail blood pressure revealed that the vasoconstrictive effect of phenylephrine was milder in TG2-/- mice leading to lower levels of lactate dehydrogenase (LDH) isoenzymes in blood. LDH isoenzyme patterns indicated more damage in lung, liver, kidney, skeletal, and cardiac muscle of wild type mice; the latter was confirmed by a higher level of heart-specific CK-MB. Our data suggest that TG2 as an α1-adrenergic receptor-coupled G protein has important regulatory functions in alpha1-adrenergic receptor-mediated metabolic processes and vascular functions.