Role of a 19S Proteasome Subunit- PSMD10(Gankyrin) in Neurogenesis of Human Neural Progenitor Cells

PSMD10(Gankyrin), a proteasome assembly chaperone, is a widely known oncoprotein which aspects many hall mark properties of cancer. However, except proteasome assembly chaperon function its role in normal cell function remains unknown. To address this issue, we induced PSMD10(Gankyrin) overexpression in HEK293 cells and the resultant large-scale changes in gene expression profile were analyzed. We constituted networks from microarray data of these differentially expressed genes and carried out extensive topological analyses. The overrecurring yet consistent theme that appeared throughout analysis using varied network metrics is that all genes and interactions identified as important would be involved in neurogenesis and neuronal development. Intrigued we tested the possibility that PSMD10(Gankyrin) may be strongly associated with cell fate decisions that commit neural stem cells to differentiate into neurons. Overexpression of PSMD10(Gankyrin) in human neural progenitor cells facilitated neuronal differentiation via β-catenin Ngn1 pathway. Here for the first time we provide preliminary and yet compelling experimental evidence for the involvement of a potential oncoprotein – PSMD10(Gankyrin), in neuronal differentiation.

Complications of veress needle insertion for creation of pneumoperitoneum

Objective: To determine the frequency of major and minor complications associated with Veress needle insertion method for creation of pneumoperitoneum for laparoscopic cholecystectomy procedure

Methods: This is a descriptive study aimed to assess the complications caused by Veress needle insertion and to determine the safety profile of this procedure. In this study patients undergoing laparoscopic cholecystectomy in Sindh Government Qatar Hospital were selected from January 2014 to December 2015. Exclusion criteria were established and those patients having age more than 80 years, procedure converted to open surgery or having previous history of upper abdominal surgeries were excluded. Complications occurring during creation of pneumoperitoneum by Veress needle were assessed. Complications were grouped as major and minor complications. Major complications included injury to abdominal viscera and vascular injuries, including injury to aorta, inferior vena cava, iliac and mesenteric arteries. Minor injuries included injury to omentum and mesentery

Results: Total of 377 cases of laparoscopic surgeries were included in the study. In these laparoscopic surgeries 3 [0.79%] injuries were recorded. All injuries recorded were categorised as minor complications and included two omental injuries and one mesenteric injury. No major complications were recorded including visceral or vascular injuries

Conclusion: It was concluded in this study that Veress needle method for creation of pneumoperitoneum has good safety profile, being less prone to major and minor complications, along with being a time-saving procedure

effect of adding body mass index [BMI] and neck circumference to Epworth sleepiness scale [ESS] for diagnosing obstructive sleep apnea hypopnea syndrome in sleep clinics

Objective: To study the effect of adding body mass index and neck circumference to Epworth Sleepiness Scale [ESS] for diagnosing obstructive sleep apnea hypopnea syndrome in sleep clinics

Study Design: Cross sectional validation study

Place and Duration of Study: Department of Pulmonology, Military Hospital Rawalpindi from 1st July 2013 to 01 September 2014

Material and Methods: The ESS and ESS plus body mass index [BMI] and neck circumference [NC] data was evaluated for 150 patients hospitalized in our hospital for polysomnographic evaluation of obstructive sleep apnea. Overnight polysomnography [PSG] was done for all patients and was considered the gold standard for diagnosis of OSA. ESS Scoring, BMI and NC data was done using pre-designated questionnaire. All the data was analyzed using SPSS version 19

Results: Age ranged from 18-74 years with mean age of the study group 53 [ +/- 12.1 SD] years. Out of study population 87 [58%] were males and 63 [42%] females. In study population 80 [55.3%] patients had ESS >10. while 102 patients had ESS>10 with BMI > 35 kg/m2 and NC>40 cm. Using an AHI >/= 5 for OSA, 136 patients [90.6%] had OSA. Sensitivity of ESS > 10 for OSA was 55.15% but increased to 72.79% when BMI > 35 kg/m2 and NC > 40 cm was added to patients with ESS > 10. Similarly specificity and predictive values of study population for diagnosing OSA also increased after adding BMI and NC to ESS

Conclusions: In this study adding body mass index [BMI] and NC to ESS score significantly increased the sensitivity and specificity and positive predictive value for diagnosis of OSA

Effectiveness of bone cleaning process using chemical and entomology approaches: time and cost

Skeletal examination is an important aspect of forensic pathology practice, requiring effective bone cleaning with minimal artefact. This study was conducted to compare between chemical and entomology methods of bone cleaning. Ten subjects between 20 and 40 years old who underwent uncomplicated medico-legal autopsies at the Institute of Forensic Medicine Malaysia were randomly chosen for this descriptive cross sectional study. The sternum bone was divided into 4 parts, each part subjected to a different cleaning method, being two chemical approaches i.e. laundry detergent and a combination of 6% hydrogen peroxide and powder sodium bicarbonate and two entomology approaches using 2nd instar maggots of Chrysomyia rufifacies and Ophyra spinigera. A scoring system for grading the outcome of cleaning was used. The effectiveness of the methods was evaluated based on average weight reduction per day and median number of days to achieve the average score of less than 1.5 within 12 days of the bone cleaning process. Using maggots was the most time-effective and costeffective method, achieving an average weight reduction of 1.4 gm per day, a median of 11.3 days to achieve the desired score and an average cost of MYR 4.10 per case to reach the desired score within 12 days. This conclusion was supported by blind validation by forensic specialists achieving a 77.8% preference for maggots. Emission scanning electron microscopy evaluation also revealed that maggots especially Chrysomyia rufifacies preserved the original condition of the bones better allowing improved elucidation of bone injuries in future real cases.

Clinical study on haemorrhoids and therapeutic evaluation of habb-e-rasaut and habb-e-muqil in its management

Haemorrhoid [Bawaseer] is a common problem characterized by an abnormally distended sub-mucosal arterio-venous shunt, embedded in swollen and inflamed anal cushion. Caused by accumulation of blood that is mixed with excess of Sauda and Balgham [Black bile and Phlegm], predisposed by improper venous drainage exacerbated by injury and inflammation. This study was done on 30 patients of 1st and 2nd degree internal haemorrhoids of either sex belonging to age group 20 to 60 years. Habb-e-Rasaut and Habb-e-Muqil were used daily for 60 days and promising results were found. Bleeding Per rectum, peri-anal pain, peri-anal itching, constipation and haemorrhoidal mass was improved in 80%, 94.4%, 90%, 85.7% and 53.3% cases respectively, after 60 days of treatment

Majoon khabsul hadeed an ultimate Unani remedy for GI bleeding disorders

The Unani drug Majoon Khabsul Hadeed was introduced by Ismail Jurjani for bleeding haemorrhoids, and with a little alteration in the formulation it is described in Ilajul Amraz, Miftah-ul-Hikmat, Qarabadeen Majeedi and Bayaz-e-Kabeer. Now this drug is available according to theformulation of Bayaz-e-Kabeer, Vol. 2, the constituents are Khabsul Hadeed Mudabbar, Oud, Saad Kufi, Zanjabeel, Filfil Siyah, Ajwain Desi, Izkhar, Halaila Siyah and Halaila Zard. In combination these constituents impart the drug Haemopoietic, Anti-Ulcerative, Ulcer healing, Styptic, Astringent, Anthelmintic and Laxative effects. These medicinal properties of the drug Majoon Khabsul Hadeed make the drug useful not only in bleeding haemorrhoids but in all bleeding conditions of gastrointestinal [GI] tract

Molecular identification of blow flies recovered from human cadavers during crime scene investigations in Malaysia

Forensic entomology applies knowledge about insects associated with decedent in crime scene investigation. It is possible to calculate a minimum postmortem interval (PMI) by determining the age and species of the oldest blow fly larvae feeding on decedent. This study was conducted in Malaysia to identify maggot specimens collected during crime scene investigations. The usefulness of the molecular and morphological approach in species identifications was evaluated in 10 morphologically identified blow fly larvae sampled from 10 different crime scenes in Malaysia. The molecular identification method involved the sequencing of a total length of 2.2 kilo base pairs encompassing the 'barcode' fragments of the mitochondrial cytochrome oxidase I (COI), cytochrome oxidase II (COII) and t-RNA leucine genes. Phylogenetic analyses confirmed the presence of Chrysomya megacephala, Chrysomya rufifacies and Chrysomya nigripes. In addition, one unidentified blow fly species was found based on phylogenetic tree analysis.

Molecular identification of blow flies recovered from human cadavers during crime scene investigations in Malaysia

Forensic entomology applies knowledge about insects associated with decedent in crime scene investigation. It is possible to calculate a minimum postmortem interval (PMI) by determining the age and species of the oldest blow fly larvae feeding on decedent. This study was conducted in Malaysia to identify maggot specimens collected during crime scene investigations. The usefulness of the molecular and morphological approach in species identifications was evaluated in 10 morphologically identifi ed blow fl y larvae sampled from 10 different crime scenes in Malaysia. The molecular identifi cation method involved the sequencing of a total length of 2.2 kilo base pairs encompassing the ‘barcode’ fragments of the mitochondrial cytochrome oxidase I (COI), cytochrome oxidase II (COII) and t-RNA leucine genes. Phylogenetic analyses confi rmed the presence of Chrysomya megacephala, Chrysomya rufifacies and Chrysomya nigripes. In addition, one unidentified blow fly species was found based on phylogenetic tree analysis.

Development of a Forensically Important Fly, Megaselia scalaris (Loew) (Diptera: Phoridae) on Cow’s Liver and Various Agar-based Diets / Jurnal Sains Kesihatan Malaysia

In forensic entomology practice, it is more common to use raw animal tissue to breed dipteran larvae and it often brings unpleasant odour in the laboratory. Few studies suggested the use of synthetic diets, mainly agar-based media, as alternatives to animal tissue but it is rarely being practiced in forensic entomology laboratory. The present study observed the growth of a forensically important fly, Megaselia scalaris (Loew) on raw cow’s liver, nutrient agar, casein agar and cow’s liver agar. A total of 100 M. scalaris eggs were transferred each into the different media and placed in an incubator at 30°C in a continuous dark condition. Data on length and developmental period were collected by randomly sampling three of the largest larvae from each rearing media, twice a day at 0900 and 1500 hours until pupariation. M. scalaris larvae reared on raw cow’s liver recorded the highest mean length (4.23 ± 1.96 mm) followed by cow’s liver agar (3.79 ± 1.62 mm), casein agar (3.14 ± 1.16 mm) and nutrient agar (3.09 ± 1.11 mm). Larval length in raw liver and liver agar were significantly different from those in nutrient and casein agar (p < 0.05). Larvae bred in liver agar and raw liver recorded the shortest larval duration before entering the post-feeding stage (89 hours), followed by nutrient agar (119 hours) and casein agar (184 hours). Total developmental time from oviposition until adult emergence for M. scalaris in liver agar and raw liver was approximately 163 hours. All puparia in nutrient agar and casein agar failed to hatch. This research highlighted the potential use of cow’s liver agar as an alternative diet of raw liver to culture M. scalaris in laboratory.

Comparative insect fauna succession on indoor and outdoor monkey carrions in a semi-forested area in Malaysia

Objective: To study the occurrence of insects fauna obtained in a semi forested area in Malaysia using the carcass of the monkey Macaca fascicularis, Raffles, and to compared the wave of insect succession collected from the carcass placed outdoor and indoor. Methods:The outdoor and indoor studies were conducted at a location of 3o17’57.86”N, 101o47’00.78”E. The euthanized monkeys were placed indoor and outdor. The insect seccession visited the carcass, the envirenmental temperature and relative humidity were recorded. Results:The main insects attracted to specific stages of decomposition were mainly members of Diptera and Coleoptera. There was a delay of fly arrival by two days in indoor carcass. Nocturnal oviposition behaviour was not observed in this study. The flies left the carcass during the twilight zone as noted during a 3 days observation period in outdoor study. The dipteran species found outdoor and indoor were similar but more diverse coleopteran species was found indoor. Conclusions:Information and evidence from this study can be used to improve the estimation of the post mortem interval in forensic cases.

Clinical manifestations and risk factors of otitis media with effusion in children presenting in tertiary care hospital

This study was conducted to find out the pattern of presentation and the risk factors associated with otitis media with effusion, in three major tertiary care hospitals of Karachi. Prospective descriptive study. This study was conducted at three major tertiary care hospital of Karachi including Civil Hospital Karachi, Lyari General Hospital and Abbasi Shaheed Hospital, Karachi, over a period of two years from November 1998 to October 2000. Two hundreds and fifty patients diagnosed as otitis media with effusion have been included in this study. Otoscopic examination, tympanic membrane mobility with Siegel's pneumatic speculum, examination of the nose and throat, pure tone audiogram and tympanometry were done in all the cases. There were 140 male and 110 female patients with mean age incidence was 7.3 [ +/- 1.2] years. Hearing impairment was the commonest symptom present in 98% of the cases. Retracted tympanic membrane was noted in 151 cases [60.4%]. Enlarged adenoids were present in 115 cases [46.0%]. Average hearing loss on pure tone audiogram was found to be 23.4 dB. Myringotomy was required in 84 cases [33.6%]. The peak age incidence of otitis media with effusion in this study was impairment with retracted tympanic membrane was the commonest clinical presentation. Enlarged adenoids was the most common and important risk factor

Serum levels of MIP-1beta and RANTES in HIV-1 subtype CRF01_AE infected patients with different rates of disease progression.

The beta-chemokines have been shown to inhibit HIV replication in vitro. To evaluate the role of serum beta-chemokines in disease progression and their anti-viral role in vivo, we determined serum levels of macrophage inflammatory protein-1beta (MIP-1beta) and regulated upon activation normal T-cell expressed and secreted (RANTES) of twenty HIV-1 subtype CRF01_AE infected patients: nine progressors (PRs, follow-up CD4+ cell count < 200/mm3 and progression to AIDS or death) and eleven slower progressors (SPs, asymptomatic and/or follow-up CD4+ cell counts > 350/mm3 at the end of follow-up) and determined their plasma viral loads. The subjects were followed for at least 36 months. All had initial CD4 values > 350 cells/mm3. In this longitudinal study, serum levels of MIP-1beta and RANTES in specimens obtained either early or later in the course of HIV infection did not differ significantly between progressors and slower progressors (p > 0.05). There were no significant changes in serum MIP-1beta and RANTES levels over time in either patient group (p > 0.05). No significant associations were observed between plasma viral loads and the measured beta-chemokines (r = -0.205, p = 0.21 for MIP-1beta and r = -0.12, p = 0.492 for RANTES). The results suggest these chemokines do not play a major systemic role in control of viremia or protection against the progression of HIV disease.

First report of the signal fly, Scholastes sp. (Diptera:Platystomatidae) visiting animal carcasses in Malaysia

Signal fly, Scholastes sp. (Diptera: Platystomatidae) was observed associated with animal carcasses in Malaysia. The first observation was on a monkey carcass, which was killed by using a handgun and immediately placed in a forested area in Gombak, Selangor while the second observation was on a pig that died of natural causes and whose carcass was placed in an oil palm plantation in Tanjung Sepat, Selangor. Both animal carcasses were visited by Scholastes sp. flies during the fresh decomposition period. However, the role Scholastes flies in the decomposition process remains unknown. In this paper, we report the occurrence of Scholastes sp. on animal carcasses in Malaysia for the first time.

The prevalence and persistence of human parvovirus B19 infection in thalassemic patients.

Human parvovirus B19 infection was studied in 60 thalassemic patients in Thailand. Seroprevalence, persistence of parvovirus B19 and their genotypes were identified in blood samples. Prevalence of anti-parvovirus B19 IgG and DNA found in thalassemic patients were 38% and 13%, respectively. Anti-parvovirus B19 IgM could be detected in 4% of these positive anti-parvovirus B19 IgG patients. The seroprevalence and parvovirus B19 DNA in patients with a history of blood transfusion were not significantly higher than those without such a history (44% vs. 34% and 20% vs. 9%, respectively). Phylogenetic analysis of NS1 nucleotide sequences of three parvovirus B19 samples revealed that they were parvovirus B19 genotype 1. They showed low genetic diversity from prototype (Au) strain. We concluded that acute and chronic persistent parvovirus B19 infection were found in the thalassemic Thai patients. Chronic persistence of parvovirus B19 infection might play important clinical role in thalassemic patients because of the high prevalence of parvovirus B19 DNA. Blood transfusion had no significant influence to increase the prevalence of parvovirus B19 infection in thalassemic patients.

Trends of HIV seropositivity at Siriraj Hospital: 13 year's observation from 1992-2004.

OBJECTIVE: The aim of the present report was to observe the trend of seroprevalence rates of HIV seropositivity for routine services at Siriraj Hospital for 13 years. MATERIAL AND METHOD: The prevalence rate of HIV seropositivity was analyzed in three groups of subjects: 1) patients who attended the hospital with HIV related diseases; 2) pregnant women at first visit to the antenatal care clinic; 3) emigrating workers who have applied for employment in foreign countries. RESULTS: Of the 13 year-observation, HIV seroprevalence rates in the groups of patients, pregnant women and emigrating workers was 10.6% (95%CI 8.9-12.3%), 2.0% (95%CI 1.8-2.2%) and 0.6% (95%CI 0.4-0.8%), respectively. CONCLUSION: The low prevalence of HIV seropositivity in the group of emigrating workers may be due to self selection, whereas the prevalence in pregnant women, which was rather consistent at about 2.0%, may represent the infection rate in the general population. The seroprevalence rate measured in the group of pregnant women demonstrates that Thailand should increase efforts to confine the spread of HIV infection in the community.

HIV-1 subtyping using gag/env heteroduplex mobility assay and peptide enzyme-linked immunosorbent assay.

Two HIV-1 subtypes have accounted for virtually all infections in Thailand: subtype B', found mainly in injection drug users (IDUs), and CRF01_AE (initially subtype E), found in over 90% of sexually infected persons and increasingly in IDUs in recent years. During 1997-1998, 227 blood samples were collected from HIV-1 infected individuals consisting of 92 mothers, 35 children and 100 IDUs. The blood samples were subtyped by heteroduplex mobility assay (HMA) and peptide enzyme-linked immunosorbent assay (PEIA). Using gag and env HMA, CRF01_AE and subtype B' accounted for 96-97% and 3-4% of both the mothers and the children, respectively. In the IDU group, 10% of the plasma samples could only be performed by gag HMA and gave the result as CRF01_AE. CRF01_AE and subtype B' using PEIA accounted for 67% and 33% of the IDUs. There was 100% concordance of the results between gag HMA and env HMA. Ninety-five percentages of concordant results were observed between HMA and PEIA. Of the 6/134 (5%) subjects with discordant results, nucleotide sequencing, used as a gold standard, confirmed the HMA result. In this study, HIV-1 was successfully genotyped by HMA and PEIA. However, a comparison of the subtyping results between HMA and PEIA revealed that HMA was slightly more accurate than PEIA.

The efficacy and effectiveness of influenza vaccination among Thai elderly persons living in the community.

OBJECTIVE: To determine the efficacy and cost-effectiveness of influenza vaccination in the Thai elderly living in an urban community. MATERIAL AND METHOD: The study design was a stratified, randomized, double blind, placebo-controlled trial. A total of 635 participants aged 60 years and older living in an urban community was randomized to receive an influenza vaccine or tetanus toxoid as a placebo injection. All participants were followed up 4-6 weeks in the community for influenza-like illness and treatment received, hospitalization and death for one year. A hemagglutination inhibition (HI) test for influenza virus antibody of all participants was done on the day of vaccination as well as 1 month, 5 months, and 12 months after the vaccination. Main outcome measures were immune response rate and protective titer, influenza-like illness, serological influenza, treatment received for influenza-like illness and their expenses, hospitalization and death during the study period. RESULTS: The immune response rate of vaccinations was 97.1% and protective titer for A (H1N1) and A (H3N2) strains were 96.4 and 98.6%, respectively. The incidence of influenza-like illness was 4.83% in the vaccine group compared with 10.88% in the placebo group. The relative risk reduction was 56% (95% CI = 14 to 77%). The survival analysis also showed that vaccinations significantly reduced the incidence of influenza (p = 0. 009). The number needed to prevent one episode was 17 persons (95% CI = 9 to 71 persons). The adverse reactions of vaccinations were mild and tolerable. However, the number of treatments received for influenza-like illness and their cost were not significantly different between the two groups. None of the subjects had pneumonia nor needed hospitalization during the study period. Seven participants died during the year of follow up, but not from influenza. CONCLUSION: In Thai elderly living in the community, influenza vaccination reduced the incidence of influenza-like illness by half, but not the number of treatments received for influenza-like illness, their cost, and its serious complications. In the year of the study, considering the cost of vaccines and the numbers needed to prevent one episode of infection from the provider's viewpoint, it may not be cost-effective to recommend that all Thai older persons living in the community should receive influenza vaccination annually. Vaccination recommendation for the elderly should be promptly implemented in expectation of a severe epidemic in Thailand.

Binding antibody to neutralizing epitope gp41 in HIV-1 subtype CRF 01_AE infection related to stage of disease.

The responsiveness of gp41 antibody against epitope ELDKWA in HIV-1 infected subjects is of importance in neutralizing viral infectivity and for being related to disease progression. In this study, antibody titers to this neutralizing epitope from HIV-1 infected subjects at asymptomatic and AIDS stages in Thailand were investigated by peptide ELISA. The results showed that the frequency of antibody production against this neutralizing epitope was low (15-35%). Moreover, antibody titers to this epitope in sera from AIDS patients were significantly lower than those in sera from asymptomatic subjects which were collected in the same year (p=0.001). Comparison between the past (1992-1994) and present (2002) sera from asymptomatic infected individuals revealed that the earlier panel contained lower antibody titers than the later panel did (p = 0.05). In addition, random sera for HIV-1 infected subjects who were infected by diverse genetic subtypes, (A through G) including CRF 01_AE, had low titers of antibody to this region as well. It is assumed that antibody production to this epitope is low and related to the stage of HIV-1 infection.

Dengue vector surveillance in urban residential and settlement areas in Selangor, Malaysia

Ovitrap surveillance was conducted in two urban residential areas (Taman Samudera Timur and Taman Samudera Selatan) and in a settlement area (Kampung Banjar), which is located 16 km from Kuala Lumpur city center, Malaysia. In Taman Samudera, dengue cases were reported monthly in 2003/2004. Thus, a study was initiated to determine the distribution and abundance of dengue vectors, Aedes aegypti and Ae. albopictus. The ovitrap surveillance indicated that Ae. aegypti and Ae. albopictus were present both indoors and outdoors. The residential sites had 73 - 79% of the ovitraps with just Ae. aegypti population and Kg. Banjar had 56% of the ovitraps with just Ae. aegypti. In the indoor and outdoor of the residential areas, together with the settlement area, the Ae. aegypti density was significantly more than Ae. albopictus (p 0.05), thus implicating that adult gravid female Ae. aegypti are present both indoors and outdoors and they do oviposit indoors and outdoors. Ae. aegypti can be incriminated as the principal dengue vector in the urban residential site, Taman Samudera and in the settlement area, Kg. Banjar.

Insecticide resistance development in Culex quinquefasciatus (Say), Aedes aegypti (L.) and Aedes albopictus (Skuse) larvae against malathion, permethrin and temephos

Laboratory-bred females of Culex quinquefasciatus, Aedes aegypti and Aedes albopictus from the insectarium, Unit of Medical Entomology, Institute for Medical Research were used in the experiment. The late third stage of the F0 larvae which survived the high selection pressure of malathion, permethrin and temephos were reared and colonies were established from adults that emerged. Cx. quinquefasciatus larvae were subjected to selection by malathion and permethrin for 40 generations, Ae. aegypti larvae to malathion, permethrin and temephos for 32 generations and Ae. albopictus larvae were selected against malathion and permethrin for 32 generations and 20 generations against temephos. The rate of resistance development was measured by LC50 value. Cx. quinquefasciatus larvae developed higher resistance to malathion and permethrin compared to Ae. aegypti and Ae. albopictus. On the whole, permethrin resistance developed at a faster rate than malathion and temephos.