METHODS: The rat embryo fibroblast (REF) cells were transfected with multi siRNA before infecting with CMV strain ALL-03. Viral growth inhibition was measured by tissue culture infectious dose (TCID50), cytopathic effect (CPE) and droplet digital PCR (ddPCR) while IE2 and DNA polymerase gene knockdown was determined by real-time PCR. Ganciclovir was deployed as a control to benchmark the efficacy of antiviral activities of respective individual siRNAs.
RESULTS: There was no significant cytotoxicity encountered for all the combinations of siRNAs on REF cells analyzed by MTT colorimetric assay (P > 0.05). Cytopathic effects (CPE) in cells infected by RCMV ALL-03 had developed significantly less and at much slower rate compared to control group. The expression of targeted genes was downregulated successfully resulted in significant reduction (P control group. Notably, the siRNAs targeting gene regions were sequenced and mutations were not encountered, thereby avoiding the formation of mutant with potential resistant viruses.
CONCLUSIONS: In conclusion. The study demonstrated a tremendous promise of innovative approach with the deployment of combined siRNAs targeting at several genes simultaneously with the aim to control CMV replication in host cells.
Materials and Methods: This study was conducted using the livers of 18 mice fixed in 10% neutral-buffered formalin. A completely randomized design with a unidirectional pattern comprising six treatments was used in this study, with each treatment consisting of three replications. Treatment 0 was the negative control group infected with P. berghei, treatment 1 was the positive control group infected with P. berghei followed by chloroquine administration at a dose of 5 mg/kg BW, and treatments 2, 3, 4, and 5 were groups infected with P. berghei and administered Malacca leaf ethanolic extracts at doses of 100, 300, 600, and 1200 mg/kg BW, respectively. The extracts were administered orally using a gastric tube for 4 consecutive days. Mice were sacrificed on the 7th day and livers were collected for histopathological examination.
Results: Histopathological examination of the livers of mice infected with P. berghei demonstrated the presence of hemosiderin, hydropic degeneration, fat degeneration, necrosis, and megalocytosis. However, all these histopathological changes were reduced in the livers of P. berghei-infected mice treated with various doses of Malacca leaf ethanolic extract. The differences between the treatments were found be statistically significant (p<0.05).
Conclusion: Ethanolic extract of Malacca leaves has the potential to protect against liver damage in mice infected with P. berghei. The dose of 600 mg/kg BW was found to be the most effective compared with the doses of 100, 300, and 1200 mg/kg BW.
Materials and Methods: In this study, grouper juveniles (92.43±standard error of the mean 0.51 mm) maintained in 31 ppt seawater were transferred into five tanks with seawater diluted to 25, 20, 15, 10, and 5 ppt. The salinity of the control group was not changed and was maintained at 31 ppt. Serum cortisol was measured using ELISA at 0, 30, 60, and 120 min after the fish were transferred to the different concentrations of salinity.
Results: The survival percentage was recorded for 14 days following the transfer and the results revealed that serum cortisol of fish in a high change in salinity (15, 10, and 5 ppt) was significantly higher than the control group immediately after exposure. At the high salinity change, the cortisol levels gradually decrease at 30 min and 60 min, until no difference in cortisol concentration was observed at 120 min. No mortality was observed in fish exposed to low salinity change (25 and 20 ppt) while in higher salinity change (5 ppt), the survival percentage was 50%.
Conclusion: The study revealed that the serum cortisol concentration was high initially and continues to decrease to resting cortisol level at 120 min indicating that cortisol hormone is released following acute stress as a primary response in grouper juveniles.
MATERIALS AND METHODS: A total of 24 ejaculates were collected from four bulls via an electroejaculator. Semen samples were diluted with 2% VCO in Tris-based extender which consists of various concentrations of SL (1, 1.25, 1.5, and 1.75%). A 20% egg yolk in Tris used as a positive control (C+). The diluted semen samples were divided into two fractions; one for chilling which were stored at 4°C for 24, 72, and 144 h before evaluated for semen quality parameters. The second fraction used for freezing was chilled for 3 h at 4°C, packed into 0.25 mL straws and then cryopreserved in liquid nitrogen. The samples were then evaluated after 7 and 14 days. Chilled and frozen semen samples were thawed at 37°C and assessed for general motility using computer-assisted semen analysis, viability, acrosome integrity and morphology (eosin-nigrosin stain), membrane integrity, and lipid peroxidation using thiobarbituric acid reaction test.
RESULTS: The results showed that all the quality parameters assessed were significantly (p<0.05) improved at 1.5% SL concentration in chilled semen. Treatment groups of 1, 1.25, 1.5, and 1.75% SL were higher in quality parameters than the control group (C+) in chilled semen. However, all the quality parameters in frozen-thawed semen were significantly higher in the C+ than the treated groups.
CONCLUSION: In conclusion, supplementation of 1.5% SL in 2% VCO Tris-based extender enhanced the chilled bull semen. However, there was no marked improvement in the frozen-thawed quality parameters after treatment.
MATERIALS AND METHODS: A total of 12 clinically healthy crossbred Boer female goats were divided into three groups; A, B and C (4 goats each per group). Group A was inoculated with 2 ml sterile phosphate buffered saline via intradermal route as the negative control group whilst Group B was inoculated with 2 ml of MA extract (1 g/ml) intradermally and Group C was then inoculated with 2 ml (1×10(9)) colony forming unit of active C. pseudotuberculosis intradermally. Blood sample was collected aseptically from the jugular vein periodically for complete blood count (CBC) analysis throughout the experimental period (3 months).
RESULT: A significant decrease (p<0.05) was observed in red blood cells, hemoglobin (Hb), packed cell volume, mean corpuscular volume and mean corpuscular Hb concentration in Groups B and C as compared to the control while WBCs, neutrophil, lymphocyte and basophil showed a significant increase (p<0.05) as compared to the control.
CONCLUSION: The inoculation of C. pseudotuberculosis and MA resulted in a significant change in the CBC, thereby, indicating that MA has a role in caseous lymphadenitis pathogenesis.
MATERIALS AND METHODS: A total of 30 healthy female boer cross goats at the age of 4 months old with average initial live body weight (BW) of 20.05±0.5 kg were used for on-farm feeding trial to evaluate the growth performance as preparation for breeding purposes. The experimental goats were divided into two groups of 15 animals each labeled as control and treatment groups, which were kept under intensive farming system. Goats in control group were fed with normal routine feeding protocol practiced by the farmer, while goats in the treatment group were fed with new feed formulation. Throughout the experimental period, on-farm monitoring and data collection were carried out. Initial BW and body condition score (BCS) were recorded before the start of the experiment while final BW and BCS were gained after 7 months of the experimental period. Average daily gain (ADG) was calculated after the experiment end. Data on BW, ADG, and BCS were recorded from both groups for every 2 weeks and reported monthly. The feed intake for the control group was 2.8 kg/animal/day which practiced by the farmer and 3.2 kg/animal/day as new feed formulation for the treatment group.
RESULTS: After 7 months of the experimental period, final BW shows an improvement in treatment group (39.1±1.53 kg) compared with control group (32.3±1.23 kg). The ADG in treatment group also gives promising result when comparing with control group. Goats in treatment group significantly attained better ADG than control group which were 126.7 g/day and 83.3 g/day, respectively. For the BCS, goats in the treatment group had shown an improvement where 86.67% (13 out of 15) of the group had BCS ≥3 (1-5 scoring scale) and only 66.67% (10 out of 15) of the control group had BCS ≥3.
CONCLUSION: Therefore, it was concluded that implementation of proper feeding program as shown in treatment group give promising result to improve the growth performance of replacement breeder goats which can be adopted by the farmers to improve farm productivity.
MATERIALS AND METHODS: About 20 quails were divided into three groups (n=8 for Groups A and B; n=4 for the control group). The quails in the Groups A and B were infected via intraocular route with 0.03 ml of 103.5 ELD50 and 107.0 ELD50 of NDV strain IBS 002, respectively, while the control group received 1× phosphate-buffered saline. Cloacal swabs and necropsy were taken on day 7 post-infection for all quails were subjected to one-step reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) for detection of virus and examination for gross pathological lesion, respectively. Blood serums of infected quails were taken on day 10, 14, and 21 post-day infections and were subjected for hemagglutination inhibition (HI) assay.
RESULTS: Depression and ruffled feathers, trachea rales, leg paralysis, and torticollis were shown in some of the quails in both infected groups. Based on statistical analysis, there was no significant difference (p>0.05) in clinical signs between the infected groups. The results for RT-qPCR were found to be negative for all groups, and no gross pathological lesions of organs observed for quails in both infected groups. Trachea, proventriculus, and cecal tonsil were taken for the detection of NDV by RT-qPCR, and some of the organ samples showed positive detection of virus in both infected groups. HI assay showed an increase in mean titers of antibody across time and between infected groups.
CONCLUSION: In summary, Japanese quails are susceptible to genotype VII NDV based on parameters assessed.
METHODS: A quasi-experimental economic evaluation comparing CPE impact on 6-month CKD mortality was conducted on the basis of payer perspective. The experimental group (n = 63) received care by health care providers who were given CPE on drug-related problems and dose adjustment. The control group (n = 80) was based on the historical cohort of patients who received care before the CPE. Measure of clinical outcome applied in this study was number of lives saved/100 patients treated. Cost-effectiveness ratios for CKD stages 4 and 5 patients without CPE and with CPE and incremental cost-effectiveness ratios (ICERs) for CKD stages 4 and 5 patients were analyzed.
RESULTS: Lives saved (%) in the treatment of CKD without CPE: CKD stage 4, 78.57; CKD stage 5, 57.58. Lives saved (%) in the treatment of CKD with CPE: CKD stage 4, 88.89; CKD stage 5, 65.45. Cost-effectiveness ratios for stage 4 with and without CPEs were Rp3,348,733.27 and Rp3,519,931.009, respectively. Cost-effectiveness ratios for stage 5 with and without CPEs were Rp7,137,874.93 and Rp7,871,822.27, respectively. ICERs were Rp2,045,341.22 for CKD stage 4 and Rp1,767,585.60 for CKD stage 5.
CONCLUSIONS: Treatment of CKD stages 4 and 5 with CPE was more effective and cost-effective compared with treatment of CKD stages 4 and 5 without CPE. The ICERs indicated that extra costs were required to increase life saved in both stages.
Materials and Methods: Twenty-four pregnant rats were randomly grouped into a control group (C), a stress group (S), and a stress group treated with TH. Eight male pups from each group were randomly chosen and they were sacrificed at eight or ten weeks of age following the novel object recognition test. Their brains were removed and histological changes and levels of MDA and NMDA receptors in the hippocampus were determined.
Results: The offspring from TH group showed significantly increased preference index (p<0.05) with higher neuronal number compared to S group. A significantly lower level of MDA and NMDA receptors were shown in TH group (P<0.01; P<0.05 respectively) compared to S group. The parameters investigated were not significantly different between C and TH groups.
Conclusion: The study has shown that memory alteration, changes in hippocampus histology, MDA and NMDA receptor levels could be prevented by TH administration during prenatal stress. The results suggest the beneficial effects of Tualang honey in prenatally stressed rat offspring.
METHODS/DESIGN: This is a randomized, single-blind, two-arm, controlled trial in patients with rheumatoid arthritis visiting outpatient rheumatology clinics in Karachi, Pakistan. We will enroll patients with established diagnosis of rheumatoid arthritis over 3 months. The patients would be randomized through a computer-generated list into the control group, i.e., usual care or into the intervention group, i.e., pharmaceutical care, in a ratio of 1:1, after providing signed written consent. The study will take place in two patient-visits over the course of 3 months. Patients in the intervention group would receive intervention from the pharmacist while those in the control group will receive usual care. Primary outcomes include change in mean score from baseline (week 0) and at follow up (week 12) in disease knowledge, adherence to medications and rehabilitation/physical therapy. The secondary outcomes include change in the mean direct cost of treatment, HRQoL and patient satisfaction with pharmacist counselling.
DISCUSSION: This is a novel study that evaluates the role of the pharmacist in improving treatment outcomes in patients with rheumatoid arthritis. The results of this trial could set the foundation for future delivery of care for this patient population in Pakistan. The results of this trial would be published in a peer-reviewed journal.
TRIAL REGISTRATION: ClinicalTrials.gov, NCT03827148 . Registered on February 2019.