Fowl adenovirus: history, emergence, biology and development of a vaccine against hydropericardium syndrome.

The poultry industry has emerged as one of the largest and fastest growing public sectors in the developed and developing countries. Unfortunately, this industry is under a major threat from diseases that are viral (Newcastle disease, infectious bursal disease, influenza, hydropericardium syndrome), bacterial (colibacillosis, pasteurellosis, salmonellosis, mycoplasmosis), parasitic (coccidiosis, histoplasmosis) or nutritional (dyschondroplasia, osteoporosis). Among these diseases, hydropericardium syndrome (HPS) is one of the important emerging diseases occurring in the specific areas of the world where broilers (chickens) are reared under intensive conditions. HPS was first observed in 1987 at Angara Goth, an area near Karachi, Pakistan, where broilers are raised. Since then, HPS has been reported in many countries of the world. From these reported cases, an adenovirus that was either isolated from or visualized electron microscopically in the liver of affected broilers has been implicated in the syndrome. The syndrome has been reproduced by inoculation of isolated fowl adenovirus (FAdV) strains, and hence, the syndrome is also called infectious hydropericardium syndrome. To our knowledge, HPS has not been observed in humans, so it is not considered a zoonotic disease, but it is of economic importance and causes huge losses to the poultry industry. Efforts have been made to develop conventional vaccines against this disease, which were formulated from infected liver homogenate. Formalin-inactivated liver organ vaccines have failed to protect the poultry industry. Hence, there is a dire need to develop a suitable vaccine to combat this disease. Currently, recombinant vaccine candidates are being developed by using molecular biology and biotechnological approaches for the prevention and control of infectious diseases, including HPS. Therefore, it is suggested that the immunogenicity of these recombinant proteins should be evaluated for their use as subunit vaccines.

How sesquiterpenes modulate signaling cascades in cancers.

Data obtained from high-throughput technologies has started to shed light on the interplay between signal transduction cascades and chromatin modifications thus adding another layer of complexity to the already complex regulation of the protein network. Based on the insights gleaned from almost a decade of research, it has now been convincingly revealed that sesquiterpenes effectively modulated different intracellular signaling cascades in different cancers. In this review we summarize how sesquiterpenes mediated Wnt, Shh, Notch and TRAIL induced signaling cascades.

Molecular cloning, expression and characterization of 100K gene of fowl adenovirus-4 for prevention and control of hydropericardium syndrome.

Fowl adenovirus-4 is an infectious agent causing Hydropericardium syndrome in chickens. Adenovirus are non-enveloped virions having linear, double stranded DNA. Viral genome codes for few structural and non structural proteins. 100K is an important non-structural viral protein. Open reading frame for coding sequence of 100K protein was cloned with oligo histidine tag and expressed in Escherichia coli as a fusion protein. Nucleotide sequence of the gene revealed that 100K gene of FAdV-4 has high homology (98%) with the respective gene of FAdV-10. Recombinant 100K protein was expressed in E. coli and purified by nickel affinity chromatography. Immunization of chickens with recombinant 100K protein elicited significant serum antibody titers. However challenge protection test revealed that 100K protein conferred little protection (40%) to the immunized chicken against pathogenic viral challenge. So it was concluded that 100K gene has 2397 bp length and recombinant 100K protein has molecular weight of 95 kDa. It was also found that the recombinant protein has little capacity to affect the immune response because in-spite of having an important role in intracellular transport & folding of viral capsid proteins during viral replication, it is not exposed on the surface of the virus at any stage.

Sampling Methods for Detection and Monitoring of the Asian Citrus Psyllid (Hemiptera: Psyllidae).

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama is a key pest of citrus due to its role as vector of citrus greening disease or "huanglongbing." ACP monitoring is considered an indispensable tool for management of vector and disease. In the present study, datasets collected between 2009 and 2013 from 245 citrus blocks were used to evaluate precision, sensitivity for detection, and efficiency of five sampling methods. The number of samples needed to reach a 0.25 standard error-mean ratio was estimated using Taylor's power law and used to compare precision among sampling methods. Comparison of detection sensitivity and time expenditure (cost) between stem-tap and other sampling methodologies conducted consecutively at the same location were also assessed. Stem-tap sampling was the most efficient sampling method when ACP densities were moderate to high and served as the basis for comparison with all other methods. Protocols that grouped trees near randomly selected locations across the block were more efficient than sampling trees at random across the block. Sweep net sampling was similar to stem-taps in number of captures per sampled unit, but less precise at any ACP density. Yellow sticky traps were 14 times more sensitive than stem-taps but much more time consuming and thus less efficient except at very low population densities. Visual sampling was efficient for detecting and monitoring ACP at low densities. Suction sampling was time consuming and taxing but the most sensitive of all methods for detection of sparse populations. This information can be used to optimize ACP monitoring efforts.

Insecticide sprays, natural enemy assemblages and predation on Asian citrus psyllid, Diaphorina citri (Hemiptera: Psyllidae).

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama is considered a key citrus pest due to its role as vector of 'huanglongbing' (HLB) or citrus greening, probably the most economically damaging disease of citrus. Insecticidal control of the vector is still considered a cornerstone of HLB management to prevent infection and to reduce reinoculation of infected trees. The severity of HLB has driven implementation of intensive insecticide programs against ACP with unknown side effects on beneficial arthropod fauna in citrus agroecosystems. We evaluated effects of calendar sprays directed against this pest on natural enemy assemblages and used exclusion to estimate mortality they imposed on ACP populations in citrus groves. Predator exclusion techniques were used on nascent colonies of D. citri in replicated large untreated and sprayed plots of citrus during the four major flushing periods over 2 years. Population of spiders, arboreal ants and ladybeetles were independently assessed. Monthly sprays of recommended insecticides for control of ACP, adversely affected natural enemy populations resulting in reduced predation on ACP immature stages, especially during the critical late winter/early spring flush. Consequently, projected growth rates of the ACP population were greatest where natural enemies had been adversely affected by insecticides. Whereas, this result does not obviate the need for insecticidal control of ACP, it does indicate that even a selective regimen of sprays can impose as yet undetermined costs in terms of reduced biological control of this and probably other citrus pests.

A subunit vaccine against hydropericardium syndrome using adenovirus penton capsid protein.

Hydropericardium syndrome (HPS) is a disease of poultry that is caused by fowl adenovirus-4. Inactivated liver homogenate from diseased birds is still the choice of vaccine in some countries which disseminates numerous pathogens along with inactivated virus. Moreover incomplete attenuation or inactivation, reversion to virulence and the oncogenic potential/genetic instability of the adenoviruses have prevented their use in routine vaccines. To address this problem an effort is made to develop a subunit vaccine. For this purpose penton base protein of HPS virus was expressed in Escherichia coli and used as subunit vaccine in broilers. Immunogenicity of the recombinant penton base protein and challenge protection test against pathogenic virus demonstrated the ability of recombinant penton base protein to confer (90%) protection. Results suggest that the recombinant penton base protein is a candidate for subunit vaccine against HPS.

Funding source and conflict of interest disclosures by authors and editors in gastroenterology specialty journals revisited.

BACKGROUND: A survey of journals published in the field of Gastroenterology conducted 5 years ago showed marked variability in reporting of conflicts of interest or funding sources in these journals. AIM: To re-examine reporting of conflicts of interest and funding sources for original articles and editorials in Gastroenterology and Hepatology journals. METHODS: We evaluated all original articles and editorials in 15 leading journals (determined by impact factor-Thomson Reuter Science Citation Index) devoted to Gastroenterology and Hepatology for disclosures of conflicts and for editor's self disclosures. We examined each journal's editorial policy by contacting the journal directly if the information was not revealed on the Web site or print versions of the journal. RESULTS: Of the 1574 articles evaluated, a total of 1207 (77%) reported the presence or absence of a potential conflict of interest and 1047 (67%) reported the presence or absence of funding sources. A total of 3 of the 15 (20%) journals (American Journal of Gastroenterology, Gastroenterology, and Alimentary Pharmacology and Therapeutics reported the presence or absence of funding sources in all their published original articles. Only 5 of 15 (33%) journals (Gut, Gastrointestinal Endoscopy, American Journal of Gastroenterology, Neurogastroenterology & Motility and Alimentary Pharmacology and Therapeutics) publicly disclosed the conflicts of interest of the editors. CONCLUSIONS: (i) Funding sources and conflicts of interest are still reported variably in the GI literature. (ii) Editorials and review articles are influential, but have poor reporting of conflicts of interest. (iii) Editors of many journals still do not report their conflicts of interest.

Essential medicines management during emergencies in Pakistan.

This paper illustrates the experiences of essential medicine management in providing cure and care to victims of Pakistan's 2005 earthquake in a safe, rational and effective mode. The health interventions assured access to essential medicine, sustained supply, inventory control through a computerized logistic support system and rational use of medicines. World Health Organization Pakistan outlined modalities for acceptance of donated medicines, assisted in speedy procurement of medicines and designed customized kits. Proper storage of medicines at controlled temperature was ensured in warehousing facilities in 12 locations. A steady supply of medicines and their consumption without stock-outs in the 56 first-level care facilities of calamity-hit areas helped to ascertain the average consumption and cost of essential medicines and supplies for the catchment population. Tools for quantification and forecasting of medicines and supplies were developed and shared. Medicines and medical supplies were efficiently used resulting in minimum wastage.

Human organ and tissue transplantation in Pakistan: when a regulation makes a difference.

Organ transplantation must be viewed in relation to the prevailing cultural, religious and economic conditions of a nation. Over the past two decades, Pakistan has emerged as one of the largest centres for commercial renal transplantation. Government efforts, supported by professional associations, civil society organizations and the media, along with World Health Organization technical assistance, have led to the development of legislation regulating this practice and curbing organ trade in conformity with international guidelines. Although only two years have passed since the enactment of the law, there is evidence that conditions have significantly improved, raising hopes for ethical and safe organ transplantation in Pakistan. This study reviews the salient features of the legislation and lists the foreseeable evolving challenges and opportunities.

Human organ and tissue transplantation in Pakistan; when a regulation makes a difference

Organ transplantation must be viewed in relation to the prevailing cultural, religious and socio-economic conditions of a nation. Over the past two decades, Pakistan has emerged as one of the largest centres for commercial renal transplantation. Government efforts, supported by professional associations, civil society organizations and the media, along with World Health Organization technical assistance, have led to the development of legislation regulating this practice and curbing organ trade in conformity with international guidelines. Although only two years have passed since the enactment of the law, there is evidence that conditions have significantly improved, raising hopes for ethical and safe organ transplantation in Pakistan. This study reviews the salient features of the legislation and lists the foreseeable evolving challenges and opportunities

Essential medicines management during emergencies in Pakistan

This paper illustrates the experiences of essential medicine management in providing cure and care to victims of Pakistan's 2005 earthquake in a safe, rational and effective mode. The health interventions assured access to essential medicine, sustained supply, inventory control through a computerized logistic support system and rational use of medicines. World Health Organization Pakistan outlined modalities for acceptance of donated medicines, assisted in speedy procurement of medicines and designed customized kits. Proper storage of medicines at controlled temperature was ensured in warehousing facilities in 12 locations. A steady supply of medicines and their consumption without stock-outs in the 56 first-level care facilities of calamity-hit areas helped to ascertain the average consumption and cost of essential medicines and supplies for the catchment population. Tools for quantification and forecasting of medicines and supplies were developed and shared. Medicines and medical supplies were efficiently used resulting in minimum wastage

Primary closure versus T-tube drainage after open choledochotomy.

BACKGROUND: Choledochotomy followed by T-tube has long been a standard surgical treatment for choledocholithiasis. It is still a preferred choice in many hospitals where minimal invasive procedures are not feasible. The use of T-tube is not without complications. To avoid the complications associated with T-tube, we have performed primary closure of the common bile duct (CBD) after exploration. This pilot study assesses the safety of primary closure of CBD, which would help form a basis for implementation on a wider scale. OBJECTIVE: To compare the clinical results of primary closure with T-tube drainage after open choledochotomy and assess the safety of primary closure for future application on a greater mass. PATIENTS AND METHODS: This comparative study was conducted at surgical unit IV Liaquat University of Medical and Health Sciences, Jamshoro, from January 2007 to December 2007. Thirty-five patients were included in the study of which 16 patients underwent primary closure. RESULTS: Thirty-five patients were included in the study. The mean age of patients who had primary closure done (n = 16) was 46.0 +/-16.8 and there were two (12.5%) males and 14 (87.5%) females. After primary closure of the CBD, bile leakage was noted in one patient (6.3%), which subsided without any biliary peritonitis as compared to the T-tube group in which two patients (10.5%) had bile leakage. Postoperative jaundice was seen in one patient (5.3%) who had a T-tube because of a blockage of CBD. Not a single patient had a retained stone in both groups as well as no recurrence of CBD stones. The postoperative hospital stay after primary closure was 5.56 +/-1.1 days as compared to after T-tube drainage which was 13.6 +/-2.3 days. The total cost of treatment in patients who underwent primary closure was USD194.5 +/-41.5 but after T-tube drainage it was USD548.6 +/-88.5. The median follow up duration for both groups was 6 months. CONCLUSION: Primary CBD closure is a safe and cost effective alternative to routine T-tube drainage after open choledochotomy.

Comparison of laboratory colonies and field populations of Tamarixia radiata, an ectoparasitoid of the Asian citrus psyllid, using internal transcribed spacer and cytochrome oxidase subunit I DNA sequences.

The genetic diversity of Tamarixia radiata Waterston (Hymenoptera: Eulophidae) laboratory colonies derived from collections in China, northern Vietnam, Pakistan, and a mixed colony from Taiwan and southern Vietnam was evaluated using the internal transcribed spacer (ITS) region 1, ITS-2, and the 5' end of the cytochrome oxidase subunit I gene. The strains share the same ITS sequence, consistent with the morphological hypothesis that the collections represent a single species. The COI marker was variable and could distinguish the northern Vietnam and Pakistan colonies from each other and from the other colonies. Comparison of COI sequences from field-collected populations of Puerto Rico, Guadeloupe, and Texas indicates that Florida is not a likely source of the introduction into Puerto Rico but is a likely source of the introduction into Texas.

Prevention of beta-thalassemia in a large Pakistani family through cascade testing.

BACKGROUND: We report cascade testing of a large Pakistani family for beta-thalassemia alleles. The family was still practicing consanguineous marriages and was at risk of having more affected births. OBJECTIVE: The objective of this study was to show that identification of disease carriers in families with index cases in order to create awareness about disease and provide genetic counseling would result in reduction of the frequency of beta-thalassemia in Pakistan. METHODS: In this large family with an index case, 27 available living members were tested for beta-thalassemia. Carriers of the disease were detected by measuring hemoglobin indices, and amplification refractory mutation system polymerase chain reaction was used for mutation analysis. Genetic counseling was provided to members of this family. RESULTS: There were already 3 marriages between the carrier members and 1 between a carrier and noncarrier in this large family; 12 (44.4%) members were found to carry the mutant gene, representing a very high carrier rate compared to the 5.4% carrier frequency of beta-thalassemia in the general population of Pakistan. The family was counseled for prevention of affected births. The initially reluctant family gradually became cooperative and seriously attended the genetic counseling sessions. CONCLUSION: Cascade testing is more practical than general population screening in a country with limited health facilities where consanguineous marriages are practiced. This report emphasizes the need of extensive testing within families with index cases to identify the carriers of beta-thalassemia in order to reduce disease occurrence through awareness and genetic counseling.

Chronomodulated drug delivery system of salbutamol sulphate for the treatment of nocturnal asthma.

A time dependent pulsed release system consisting of an effervescent core surrounded by consecutive layers of swelling and rupturable polymers was prepared and evaluated. The cores containing salbutamol sulphate as bioactive agent were prepared by direct compression method using different ratios of microcrystalline cellulose and effervescent agent and then coated sequentially with an inner swelling layer containing a hydrocolloid, hydroxypropylmethylcellulose E5 and an outer rupturable layer having Eudragit RL/RS (1:1). The effects of various processing and formulative parameters on the performance of system were studied. The rupture and dissolution tests were studied using the USP paddle method at 50 rpm in 0.1 N HCl and phosphate buffer pH 6.8. The lag time of the drug release decreased by increasing the inner swelling layer and increased by increasing the rupturing layer level. All the results obtained in the present study suggest that osmotic pumping effect was involved which eventually lead to the drug release.

Identification of hepatitis B virus core mutants by PCR-RFLP in chronic hepatitis B patients from Punjab, Pakistan.

Chronic hepatitis B virus (HBV) infection remains a major health issue worldwide. Several factors including core gene variation are responsible for the development of chronicity of HBV infection. The present study was designed to identify the variations in the core region of the HBV genome in a local population of chronic hepatitis B patients (n = 57) using a PCR-based restriction fragment length polymorphism (PCR-RFLP) method. Fifty subjects were found to be positive for the presence of HBV DNA. For the core region genotyping, the Ava II and Msp I restriction enzymes were used. Mutations at nucleotide (nt) 2147 and nt 2362 in the HBV genome in the core region for Ava II (A4 type, 74%) and nt 2331 for Msp I (M1 type, 66%) were observed as the most common pattern. These results are different from those of previously reported studies on other populations and thus appear to be unique to the Pakistani population. This type of characterization of core mutants may be useful for the design of vaccines based on viral epitopes that are effective for the Pakistani population. Moreover, these unique genotypic patterns for the HBV core gene might be some of the main factors responsible for understanding the underlying mechanism by which HBV chronicity is developed in the Pakistani population.

Positive correlation of fitness with group size in two biotypes of Russian wheat aphid (Homoptera: Aphididae).

Changes in fitness parameters as a function of colony size (one versus 10 aphids) were measured in two biotypes (RWA1 and RWA2) of the Russian wheat aphid, Diuraphis noxia (Mordvilko) (Homoptera: Aphididae), feeding on three cultivars of wheat, Triticum aestivum L., at two temperatures. 'Trego' is a cultivar with no specific resistance to D. noxia, whereas, 'Stanton' and 'Halt' express Dny and Dn4 resistance sources, respectively. Feeding in a group accelerated the development of RWA1 on Trego and Stanton at 20 degrees C, but not at 24 degrees C, whereas grouped RWA2 developed faster than solitary RWA2 on all three cultivars at 24 degrees C, but not at 20 degrees C. Survival (first instar-adult) of RWA2 also was improved by grouping on Stanton and Halt at 24 degrees C, but solitary RWA2 survived better at 20 degrees C on all three cultivars. The reproductive rate of RWA1 was improved by grouping on Trego and Stanton at both temperatures, but only on Halt at 24 degrees C. Lifetime fecundity of RWA1 also was increased by grouping in all cases except for Trego at 20 degrees C. Grouped development increased the reproductive rate of RWA2 on all three cultivars at 24 degrees C, but had no effect at 20 degrees C. Grouped RWA2 developed and reproduced faster than grouped RWA1 on all three cultivars at 24 degrees C. Thus, the fitness of D. noxia was positively correlated with group size during colony establishment, but the effects were sensitive to temperature, being more pronounced at 20 degrees C for RWA1 and at 24 degrees C for RWA2.

Can renal scan findings predict biopsy-proven allograft rejection?

OBJECTIVES: To assess the usefulness of isotopic renogram in diagnosing acute renal graft rejection. MATERIALS AND METHODS: Degree of perfusion and allograft uptake of tracer were correlated with the clinical and biopsy diagnoses in 15 postrenal transplant patients with varying degrees of renal impairment. Renographic findings and perfusion calculations were done by a blinded observer. RESULTS: A strong correlation was found between renal histology and renal scan findings in 13 of 15 patients. Sensitivity and specificity of renal scanning in diagnosing acute rejection were 85% and 50% respectively (using renal biopsy findings as the gold standard). CONCLUSION: Our results demonstrate a strong correlation between blinded perfusion assessment and biopsy-proven acute rejection. We conclude, therefore, that single renal flow scan with DTPA (noninvasive/nonnephrotoxic) allows a physician to tailor therapy for acute renal graft dysfunction. We suggest that in cases with a renographic diagnosis of AR, the patient should receive standard antirejection therapy. Renal biopsy should be reserved for those instances when the renographic findings are not definitive and those when the patient fails to respond to a standard methylprednisolone therapy.

Pregnancies in renal transplant recipients--with a focus on the maternal issues.

One hundred and thirteen pregnancies in 73 women who received a renal transplant were studied. The duration between the transplant operation and the conception averaged 19.9 months with a range of I month to 72 months. The mean age was 28 years. The majority (50 recipients had one pregnancy each, but two women had 5 children each and one had seven children). Only 12 (11.5%) out of the 113 pregnancies ended up with spontaneous abortions. The maternal medical problems encountered were reversible rejection in 11%, hypertension in 43%, UTI in 17% and gestational diabetes in 21%. It was noted that there was high incidence of preterm delivery in 64% of the pregnancies Caesarian section was required in 72% % of the cases. In conclusion we observed a high prevalence of successful pregnancies with no adverse effect on the allograft function and excellent fetal outcome (which is described in a separate paper focusing on the fetal outcome).

Pregnancies in renal transplant recipients--with a focus on babies.

We studied 113 pregnancies in 72 renal transplant recipients. The mean duration between the transplant operation and the first pregnancies was 19.9 months (1-72). This paper focuses on the findings in babies and their outcome. The male babies were 34% of the total. 28% of the babies were delivered vaginally and 72% by Caesarian section. It was noted that there was high incidence of preterm delivery in 64% of the pregnancies. The birth weights were also less than that observed in the general population. 84% was under 50th percentile for weight with 19% being under the 10th percentile. (Saudi growth chart, KACST) The mean hospital stay was 18 days (2-44). The Apgar score was less than 7 in only 10% of the cases and 10 in 50.9%. The incidence of congenital defects was low and no different than the general population (we encountered only 4 cases with minor defects). Despite exposure to cyclosporine throughout the pregnancy and having reduced Nephron mass by virtue of their low birth weight we could not find any glomerular or tubular defects, hypertension or prorienuria in 41 children with a mean age of 52 months. We advised against breast feeding in view of the presence of cyclosporine in the mothers' milk.