Development of male sterility by silencing Bcp1 gene of Arabidopsis through RNA interference

The development of male sterility is one of the most important steps in hybrid seed production. Several methods for the abortion of pollens based on conventional as well as genetic engineering are reported for the various crop species. Here we have investigated the use of RNA interference (RNAi) technology to silence a male specific gene, Bcp1 in the model host Arabidopsis thaliana. Bcp1 is active in both diploid tapetum and haploid microspores. Three batches of explants (A. thaliana) were selected onherbicide glufosinate ammonium and putative transgenes were confirmed through PCR and Southern hybridization. About 49 out of 58 Arabidopsis lines transformed with RNAi construct containing Bcp1 sequences were male sterile. Transgenic plants were phenotypically indistinguishable from nontransgenic plants and by crossing with non-transgenic fertile pollens successful seed set was observed. The Bcp1 gene was also amplified from chilies, tomato and brassica. The present study resulted in developing male sterile A. thaliana (Eco. Columbia) line through genetic engineering. The diversity of Bcp1 gene will help us to produce transgenic male sterile plants in crop plants

PREPARATION OF CONJUGATE FOR USE IN AN ELISA FOR HUMORAL IMMUNE RESPONSE AGAINST EGG DROP SYNDROME VIRUS IN LAYER CHICKS

An indirect enzyme-linked immunosorbent assay (ELISA) was performed for the detection of antibodies against Egg Drop Syndrome (EDS) virus. Virus identification was done through haemaggluti- nation inhibition (HI) test using known antisera. Antichicken immunoglobulins were raised in goats and purified by ammonium sulphate precipitation technique. These goat-antichicken immunoglobulins were conjugated with horseradish peroxidase. Twenty-seven serum samples were collected from a layers flock vaccinated against EDS and specific antibodies were determined by using a horseradish conjugate

Molekularna filogenija i karakterizacija mundri ovce (Ovis aries) u Pakistanu sekvenciranjem mitohondrijskog citokroma b i podjedinice I citokrom oksidaze

The main focus of this research is to determine the molecular phylogeny and characterization of Mundri Sheep (Ovis aries) through sequencing of mitochondrial Cytochrome b and Cytochrome Oxidase Subunit I (COI). This sheep breed appears morphologically different from other local sheep breeds of Pakistan. The current research is carried out to appraise the status of Mundri sheep whether it is aa different breed from other breeds or not. Blood samples of Mundri sheep were collected from Livestock Experiment Station, (LES) Fazilpur in district Rajanpur (Punjab). DNA was isolated and subjected to Polymerase Chain Reaction (PCR) for amplification of Cytochrome b and COI genes using appropriate primers. PCR products were sequenced and analyzed by MEGA X software. The phylogeny analysis categorized Ovis aries including Mundri sheep into three and two groups for Cytochrome b and COI genes respectively. It showed Mundri sheep as a separate group and thus as a separate breed from all other local sheep breeds. Hence the study validates based on Cytochrome b and COI that Mundri sheep is a distinctive breed from the rest of the local sheep breed.Glavni je cilj ovog istraživanja bio odrediti molekularnu filogeniju i karakterizaciju mundri ovce (Ovis aries) sekvenciranjem mitohondrijskog citokroma b i podjedinice I citokrom oksidaze (COI). Ova se pasmina ovaca morfološki čini drugačijom od ostalih lokalnih pasmina ovaca u Pakistanu. Ovo je istraživanje provedeno da bi se procijenio status mundri ovce, da bismo mogli odrediti radi li se o pasmini drugačijoj od ostalih pasmina. Uzorci krvi mundri ovce prikupljeni su iz Stanice za eksperimente na stoci (engl. Livestock Experiment Station – LES) Fazilpur u okrugu Rajanpur (Punjab). DNK je izolirana i podvrgnuta lančanoj reakciji polimerazom (PCR) zbog pojačanja citokrom B i COI gena uporabom prikladnih primera. PCR proizvodi su sekvencirani i analizirani pomoću MEGA X softvera. Filogenetska analiza kategorizirala je Ovis aries uključujući mundri ovcu, u tri i dvije skupine za citokrom b, odnosno COI gene. Istraživanje je pokazalo da je mundri ovca posebna skupina i time zasebna pasmina ovaca u odnosu na ostale lokalne pasmine. Na temelju citokroma b i COI, naša je studija potvrdila da je mundri ovca zasebna pasmina i da se razlikuje od ostalih lokalne pasmine ovaca

Effectiveness of an antimicrobial treatment scheme in a confined glanders outbreak

BACKGROUND: Glanders is a contagious and fatal zoonotic disease of solipeds caused by the Gram-negative bacterium Burkholderia (B.) mallei. Although regulations call for culling of diseased animals, certain situations e.g. wild life conservation, highly valuable breeding stock, could benefit from effective treatment schemes and post-exposure prophylaxis. RESULTS: Twenty three culture positive glanderous horses were successfully treated during a confined outbreak by applying a treatment protocol of 12 weeks duration based on the parenteral administration of enrofloxacin and trimethoprim plus sulfadiazine, followed by the oral administration of doxycycline. Induction of immunosupression in six randomly chosen horses after completion of treatment did not lead to recrudescence of disease. CONCLUSION: This study demonstrates that long term treatment of glanderous horses with a combination of various antibiotics seems to eliminate the agent from the organism. However, more studies are needed to test the effectiveness of this treatment regime on B. mallei strains from different endemic regions. Due to its cost and duration, this treatment can only be an option in certain situations and should not replace the current “testing and culling” policy, in conjunction with adequate compensation to prevent spreading of disease

Post Total Thyroidectomy Hypocalcaemia and relation with the Age of the Patient

Objective: To find the magnitude of clinically overt hypocalcaemia post-total thyroidectomy and its relation with the age of the patient. Study Design: Cross-sectional study. Place and Duration of Study: Combined Military Hospital, Bahawalpur Pakistan, from Jan 2017 to Dec 2019. Methodology: Sixty patients who were to undergo total thyroidectomy were included in the study in strict compliance with the inclusion and exclusion criteria. Patients were divided into two groups according to their ages. In Group-A, patients younger than 50 and Group-B, patients older than 50 years were included. All the patients were followed up by clinical examination for the development of clinically overt hypocalcaemia and confirmed by measurement of serum calcium. Results: Mean age of the patients was 42.75±13.05 years, with an age range of 20 to 68 years. Of all the patients, 14(23.3%) were male, and 46(76.7%) were females. Seven patients in Group-A developed post-thyroidectomy hypocalcaemia clinically,whereas, in Group-B, only one developed this condition. Clinical post-thyroidectomy hypocalcaemia developed more often in the younger age group and these results were statistically significant (p= 0.023). Conclusion: Clinically overt hypocalcaemia develops more often in younger patients after total thyroidectomy. Keywords: Biochemical hypocalcaemia, Clinical hypocalcaemia, Post thyroidectomy hypocalcaemia, Total thyroidectom

Optimal modeling of spindle-tool system with design variables using Taguchi based approach

A generalized finite element (FE) model of the spindle machine based totally on Timoshenko’s beam idea is hired to discretize the meeting with joint interfaces idealized with the aid of contact spring models and the model is confirmed with experimental modal analysis at the spindle-tool system. The effects of five important variables namely tool overhang, bearing span,tool diameter, contact stiffness,and axial preload are simultaneously studied on the dynamic response of the spindle-tool unit.Using method ofdesign of experiments(DOE),the dynamic stiffness data along with fundemental natural frequencies are recorded for different combinations of design variables. Initially after studying the parametric effects using analysis of variance(ANOVA)

The Merging of Two Dynasties—Identification of an African Cotton Leaf Curl Disease-Associated Begomovirus with Cotton in Pakistan

Cotton leaf curl disease (CLCuD) is a severe disease of cotton that occurs in Africa and Pakistan/northwestern India. The disease is caused by begomoviruses in association with specific betasatellites that differ between Africa and Asia. During survey of symptomatic cotton in Sindh (southern Pakistan) Cotton leaf curl Gezira virus (CLCuGV), the begomovirus associated with CLCuD in Africa, was identified. However, the cognate African betasatellite (Cotton leaf curl Gezira betasatellite) was not found. Instead, two Asian betasatellites, the CLCuD-associated Cotton leaf curl Multan betasatellite (CLCuMB) and Chilli leaf curl betasatellite (ChLCB) were identified. Inoculation of the experimental plant species Nicotiana benthamiana showed that CLCuGV was competent to maintain both CLCuMB and ChLCB. Interestingly, the enations typical of CLCuD were only induced by CLCuGV in the presence of CLCuMB. Also in infections involving both CLCuMB and ChLCB the enations typical of CLCuMB were less evident. This is the first time an African begomovirus has been identified on the Indian sub-continent, highlight the growing threat of begomoviruses and particularly the threat of CLCuD causing viruses to cotton cultivation in the rest of the world

Managing Chronic Diseases in Family Medicine: Best practices and Evidence-Based Approaches

The management of chronic diseases within the realm of family medicine presents a multifaceted challenge with profound implications for healthcare systems and patients alike. Chronic diseases, such as diabetes, hypertension, and cardiovascular conditions, are prevalent and impose a significant burden on individuals, families, and society as a whole. This article explores best practices and evidence-based approaches for managing chronic diseases in family medicine. It delves into the epidemiological landscape of chronic illnesses, emphasizing the need for effective prevention and management strategies. Evidence-based Models, such as The Chronic Care Model (CCM), Patient-Centered Medical Home (PCMH), and Self-assessment models are discussed in the context of family medicine. The importance of comprehensive, coordinated, and patient-centric approaches is underscored, highlighting the pivotal role of primary care physicians in the ongoing battle against chronic diseases. It is clear, that development in the field of family medicine underscores the importance of patient involvement in diseases management process through shared-decision making model. Although such model require physicans to spend more time educating patients so they can make informed decisions and implement self-management strategies, it has overall better health outcomes and eventually needs to requiring less intervention by physicians

Seroprevalence of hepatitis C in type 2 diabetes: evidence for a positive association

<p>Abstract</p> <p>Background</p> <p>There is a growing body of literature on the relationship of Hepatitis C virus infection (HCV) and type 2 diabetes mellitus (T2DM). However, there are certain gaps in literature and the data is inconclusive. This study was, therefore, carried out to determine the prevalence of HCV infection in diabetic patients and to elucidate the presence of any possible relationship between HCV and T2DM in this region.</p> <p>Methods</p> <p>Serologic testing for anti-HCV antibody was done on a sample of 3000 individuals with T2DM visiting Diabetes Clinic of Nishtar Medical College Hospital, Multan and 10,000 volunteer blood donors visiting blood bank of the same hospital during the study period using Accurate rapid immunochromatographic kits which was later confirmed by using Chemelex S.A third generation ELISA kit for positive cases. Data about various variables was collected from diabetic patients using a structured questionnaire after taking informed consent.</p> <p>Results</p> <p>Prevalence rate of 13.7% for HCV infection was recorded among subjects having T2DM with seropositivity rate of 4.9% among the control group of volunteer blood donors without diabetes. The patients with T2DM were more likely to have HCV infection as compared to the control group (OR = 3.03, 95%CI = 2.64-3.48, p = 0.001). Diabetic patients with age above 55 years had higher prevalence rate as compared to younger individuals. Male patients had significantly high seropositivity as compared to female patients (15.3% vs. 12.4%, p = 0.02). Those with duration of diabetes 11 years and above and the ones with good glycemic control had higher seroprevalence rates of 18.2% and 18.7% respectively. There was no statistically significant difference among subjects when the distribution of HCV was studied on the basis of marital status, locality, or family history of diabetes.</p> <p>Conclusions</p> <p>The results show that there is a strong association between HCV and T2DM in the region as evident from significantly higher prevalence of HCV infection in diabetics as compared to the control group in the present study.</p