Expression of hepatitis B virus chimeric proteins in prokaryotic and eukaryotic systems

The particulate form of the core antigen of hepatitis B virus (HBcAg), is highly immunogenic. It has been used as a carrier molecule, for expression and presentation of heterologous viral epitopes on the surface of hybrid core particles, in immunogenicity studies. The aim of this project was to produce a hybrid antigen comprising HBcAg and an immunogenic epitope of human cytomegalovirus (HCMV). A direct comparison was made of amino and carboxyl terminal fusions, by investigating the influence of position of the foreign epitope on antigenicity, immunogenicity and hybrid core particle formation. A part of the HCMV genome, encoding a neutralizing glycoprotein epitope gp58, was inserted at the amino terminus or fused to the truncated carboxyl terminus of HBcAg in separate constructs and expressed in a prokaryotic system (E.coli). At the same time, in order to express the same fusion proteins in a eukaryotic system, the baculovirus expression vector system (BEVS) was selected and as an initial control two recombinant baculoviruses, containing genes encoding HBcAg and hepatitis B surface antigen were isolated by dot blot hybridization. It was realized that work in BEVS would require more time than previously expected therefore further work was only carried out on the prokaryotic system. The HBcAg carboxyl terminal fusion (HBC 3 - 1 4 4 -HCMV) was expressed in high yields in E.coli and assembled into core like particles resembling native HBcAg. A similar fusion in the amino terminus of HBcAg (HCMV-HBC1-183) could not be purified or characterized immunologically, although it formed core like particles. HBc3-144-HCMV displayed HBc antigenicity but HCMV antigenicity could not be detected. Following immunization of rabbits with HBC3-144-HCMV, a high level of anti-HBc specific antibody was produced along with HCMV/gp58 specific antibody. The data presented here provide evidence that the HCMV/gp58 region can be used as a candidate immunogen for an HCMV subunit vaccine and that HBcAg can effectively present this foreign epitope joined to its carboxyl terminus, to the immune system

FREQUENCY OF ENCEPHALOPATHY IN CHILDREN WITH ENTERIC FEVER

Objective; To determine the frequency of encephalopathy in children with enteric fever presenting at a tertiary care hospital. Patients and methods; This descriptive, cross-sectional study was conducted at Department of Pediatric Medicine Services Hospital Lahore from January 2015 to December 2016. A total of 100 patients with enteric fever were recruited in our study. Results; Of these 100 study cases, 59 (59%) were boys and 41 (41%) were girls. Mean age of our study cases was 7.99 ± 3.28 years. Majority of the study cases with enteric fever i.e. 56 (56%) were age more than 7 years. Mean Temperature was 102.30 ± 0.45 F0. Mean disease duration (enteric fever) was 5.47 ± 1.18 days (with minimum disease duration was 4 days while maximum duration of fever was 8 days). Of these 100 study cases, 51 (51%) belonged to the rural areas and 49 (49%) were from urban territories. While majority of these children belonged to the families having poor socio-economic status 67 (67%), middle income socio-economic status was reported by 26 (26%) and 07 (7%) were from higher socio-economic status. Encephalopathy was seen in 18 (18%) of our study cases. Conclusion; Our study results have indicated that frequency of encephalopathy is high among children hospitalized due to enteric fever. Majority of our study cases belonged to poor socio-economic status families living in slum areas where poor sanitation facilities. Encephalopathy was significantly associated with disease duration, increasing age in male gender and lower age groups in female gender. Keywords; Enteric fever, Encephalopathy, children

COPPER DEFICIENCY IN RUMINANTS IN PAKISTAN

Copper (Cu) is a vital micronutrient in all living organisms. It is essential for the proper functioning of the different enzymes like ceruloplasmin, dopamine β, superoxide dismutase, cytochrome c oxidase, lysyl oxidase, monoamine oxidase, tyrosinase, peptidylglycine α, glactose oxidate, ascorbate oxidase and monooxigenase. These enzymes play a vital role in a variety of physiological functions of hematological, nervous, cardiovascular, reproductive, antioxident and immune systems. Hypocuprosis may be due to primary or secondary copper deficiency. The main reason of copper deficiency in Pakistani livestock is most probably due to the presence of dietary Cu antagonist, e.g sulphur and in particular molybdenum. Clinical symptoms of copper deficiency include ataxia, paralysis, paresis, incoordination, head tremors, edema, twisted joints, weakness of limbs, poor quality fleece, bespectacled eyes, fractures, anemia, depigmentation of hair and skin, rough body coats, thin bellies, degeneration of myocardial muscles, ataxia, head tremors and falling disease. In Pakistan copper has been used for the treatment of different health disorders of livestock e.g nutritional hemoglobinuria, parturient hemoglobinuria, mastitis, anestus, leukoderma, chronic diarrhea

Geographic-Based Spray-and-Relay (GSaR): An Efficient Routing Scheme for DTNs

In this article, we design and evaluate the proposed Geographic-based Spray-and-Relay (GSaR) routing scheme in Delay/Disruption Tolerant Networks (DTNs). To the best of our knowledge, GSaR is the first spray based geographic routing scheme using the historical geographic information for making routing decision. Here, the term spray means only a limited number of message copies are allowed for replication in the network. By estimating a movement range of destination via the historical geographic information, GSaR expedites message being sprayed towards this range, meanwhile prevents that away from and postpones that out of this range. As such, the combination of them intends to fast and efficiently spray the limited number of message copies towards this range, and effectively spray them within range, in order to reduce the delivery delay and increase the delivery ratio. Furthermore, GSaR exploits Delegation Forwarding (DF) to enhance the reliability of routing decision and handle the local maximum problem, considered as the challenges for applying geographic routing scheme in sparse networks. We evaluate GSaR under three city scenarios abstracted from real world, with other routing schemes for comparison. Results show that GSaR is reliable for delivering messages before expiration deadline and efficient for achieving low routing overhead ratio. Further observation indicates that GSaR is also efficient in terms of a low and fair energy consumption over the nodes in the network

Surgical management of traumatic extra dural hematoma in children: Experiences and analysis from 24 consecutively treated patients in a developing country

Background: Children with epidural hematoma (EDH) present differently thanadults. The outcome of treatment is also different. We aim to report our experiences with EDH in pediatric age group in terms of mode of injury, presenting features, management, and outcomes. We also aim to identify different prognostic indicators in pediatric patients with EDH. Methods: We prospectively collected data from 24 consecutively surgically treated pediatric patients. The data collected included presenting features, radiological imaging, details of management, and outcomes. Descriptive analysis was performed and different variables were tested for any statistical significance with Glasgow Outcome Score (GOS). Results: There were 19 male and 5 female patients. The mean GlasgowComa Scale (GCS) score at presentation was 9.3 ± 4.4. Falls were the mostcommon cause of EDH. Outcome assessment was done at 3 month follow up.A total of 15 patients had a GOS score of 5, 4 patients had a GOS score of4, 2 patients had a GOS score of 3, while 3 patients had a GOS score of 1.On univariate analysis, admitting GCS score, patient’s age, the time frominjury to admission and injury to surgery, anisocoric pupils at presentation andeffacement of basal cisterns were significantly associated with the outcome ofGOS score. Conclusion: Falls are the most common mode of injury leading to EDH inchildren. Lower GCS at presentation, younger age at trauma, increased timesince trauma to surgery and admission, anisocoria and effacement of basalcisterns are statistically significant variables in surgically treated pediatric patients of EDH that confer a poorer prognosis. A timely surgical intervention can result in excellent outcomes

Innovations in MDR therapy: essential oil applications

There are three common outcomes when testing the application of combinatory therapies for potential use in the clinical setting against multi-drug resistant (MDR) bacteria; whether synergistic, additive or antagonistic. Due to their relatively more favourable outcomes, antibiotics and adjuvant achieving synergism have been the primary focus in the development of therapeutics thus far; this has also resulted in less attention given to additive interactions, which may be equally effective. Hence, in this study, the authors investigated the effects of additivity using a representative combination model involving Cinnamomum verum (cinnamon bark) essential oil (CBO) and meropenem, a carbapenem class of antibiotics. Based on the zeta potential measurements, outer membrane permeability and scanning electron microscopy, synergistic and additive interactions of cinnamon bark oil (CBO) indicated comparable bacterial membrane disruption, alluding similar degrees of cell perturbations at the molecular level. Results from this study suggests that further investigations should be looked into for combinatorial therapeutic pairs achieving additive indices so as not to disqualify potentially useful alternatives in antibiotic therapy

Health systems changes after decentralisation: Progress, challenges and dynamics in Pakistan

Decentralisation is widely practised but its scrutiny tends to focus on structural and authority changes or outcomes. Politics and process of devolution implementation needs to be better understood to evaluate how national governments use the enhanced decision space for bringing improvements in the health system and the underlying challenges faced. We use the example of Pakistan\u27s radical, politically driven provincial devolution to analyse how national structures use decentralisation opportunities for improved health planning, spending and carrying out transformations to the health system. Our narrative draws on secondary data sources from the PRIMASYS study, supplemented with policy roundtable notes from Pakistan. Our analysis shows that in decentralised Pakistan, health became prioritised for increased government resources and achieved good budgetary use, major strides were made contextualised sector-wide health planning and legislations, and a proliferation seen in governance measures to improve and regulate healthcare delivery. Despite a disadvantaged and abrupt start to devolution, high ownership by politicians and bureaucracy in provincial governments led to resourcing, planning and innovations. However, effective translation remained impeded by weak institutional capacity, feeble federal-provincial coordination and vulnerability to interference by local elites. Building on this illustrative example, we propose (1) political management of decentralisation for effective national coordination, sustaining stable leadership and protecting from political interfere by local elites; (2) investment in stewardship capacity in the devolved structures as well as the central ministry to deliver on new roles

Diabetes Score questionnaire for lifestyle change in patients with type 2 diabetes

Background. Designed for use in clinical settings, the Diabetes Score is a 10-item, one-page questionnaire for discussing lifestyle change. We aimed to evaluate the Diabetes Score questionnaire for its validity and acceptability among individuals with type 2 diabetes.Methods. An observational study was conducted using interviewer-administered questionnaires to adult patients with type 2 diabetes at three ambulatory clinics. We used the Diabetes Score questionnaire for measuring adherence to diet, exercise and other lifestyle recommendations. The questionnaire yields an intuitive score ranging from 0 to 100, by addition of each of the 10 items which are rated as 0, 5 or 10 by the patient. A score of more than 60 was consideredsatisfactory.Results. A total of 311 patients, 56% females, with a median age of 55 years (range: 23 to 87) participated in the study. Diabetes Score correlated with glycemic control, HbA1c (r = –0.20) and blood glucose (r = –0.25; P < 0.001), indicating validity. Reliability was demonstrated by internal consistency (alpha .577) and discriminant factor analysis. Based on multivariate modeling, an improvement of 30 points on the Diabetes Score corresponded to a drop in HbA1c by 1.0%-unit (11 mmol/mol).Conclusion. Diabetes Score is a valid and reliable tool for empowering lifestyle and behavior modification among patients with diabetes mellitus. This brief and free-to-use questionnaire has the potential to be used in diabetes clinics to discuss behavior change. It can serve as the first-line intervention in diabetes patients while reducing the cost of diabetes care