Cloning and periplasmic soluble expression of hepatitis B surface antigen gene in Escherichia coli.

OBJECTIVE: The objective of this study was to clone and express the hepatitis B surface antigen gene (HBsAg) in Escherichia coli (E. coli), thereby aiming to develop potential local therapeutics for combating Hepatitis B virus (HBV) infection in the Pakistani community by producing HBsAg in E. coli. MATERIALS AND METHODS: Blood serum samples were collected from hepatitis B-infected patients, and their genomic DNA was extracted. Real-time and nested polymerase chain reaction (PCR) was performed to amplify the HBsAg gene. The gene of interest was cloned into the pET20b expression vector and transformed into E. coli BL21 (DE3) using Isopropyl ß-D-1-thiogalactopyranoside (IPTG) induction. The gene's precise size was confirmed with gene-specific external and internal primers (681 bp and 400 bp, respectively). RESULTS: The HBsAg gene was successfully sequenced and submitted to GenBank, exhibiting 98% homology with targeted HBV sequences worldwide. The expression of HBsAg protein was confirmed through silver staining, Coomassie staining, western blot, and dot blot analysis. CONCLUSIONS: The expressed protein clones are now available for further development as a local recombinant DNA vaccine to prevent hepatitis B viral infection in the local community.

Distribution, host range and toxicity assessment of different insecticides on Bactrocera diversa Coquillett, 1904 (Diptera: Tephritidae).

The present study was conducted to investigate the array of hosts, distribution and to evaluate the toxicity of four insecticides: imidacloprid, fipronil, cypermethrin and chlorpyrifos alone and in combination against 3rd instar maggot and adult stage of fruit fly Bactrocera diversa Coquillett, 1904 (Diptera: Tephritidae) during 2021. B. diversa maggots were found vigorously feeding inside the cucurbit hosts (pumpkin, cucumber, bitter gourd, watermelon, round melon, bottle gourd) collected from different localities of Poonch division of Azad Jammu & Kashmir, Pakistan, and this species is reported for the first time as new record to this region. Susceptibilities of B. diversa to insecticides were evaluated using topical method. Mortality was checked after 3, 6, 8 and 24h of exposure. Cypermethrin was most effective to kill 50% of both larval and adult stage with least LC50 [7.2(1.040±0.214), 17.4(0.748±0.193)], respectively followed by imidacloprid. Imidacloprid most effectively killed 90% of both larval and adult population with least LC90 value [73.2 (3.013±0.708) 16.9 (1.886±0.437)] respectively after 24 hours. Cypermethrin with chlorpyrifos most effectively killed 50 and 90 percent of both larval and adult stage of B. diversa with least LC50 value [11.3 (1.085±0.245), 2.5 (0.759±0.252)] and least LC90 value [171.3 (1.085±0.245), 121.9 (0.759±0.252)], respectively after 24h of exposure. Toxicity of each insecticide increased with exposure for longer time and increased dose. Cypermethrin is suggested as most effective against both larval and adult stages of B. diversa in combination with chlorpyrifos followed by imidacloprid.

Clinical audit on indications and technique of male urethral catheterization in emergency department of a tertiary care hospital.

Urethral catheterization (UC) is a common procedure done in the Emergency Department of a hospital. The main indications for a UC are relieving acute urinary retention and bladder outlet obstruction. This cross-sectional audit was carried out between July 2019 to February 2020, in the Emergency Department (ED). All male patients aged 18 years and above, in whom UC was performed, were included in the audit. A total of 81 male patients were surveyed. Although, sterilized technique was maintained by using sterile gloves and pyodine but hand hygiene (sterilization or hand wash) was not performed before the procedure 80% of times. CDC hand hygiene guidelines were distributed to promote hand hygiene. What do we already know about this topic ? The Lippincott guidelines are considered gold standard which includes hand hygiene as an important element of urinary catheterization UC. The CDC Center of Disease Control CDC promotes the use of mandatory gloves and generous hand sanitizing before procedures. How does your research contribute to the field For continuous improvement and patient focused practice, we must audit the technique of nursing staff, doctors and health care assistants who are involved in insertion and ordering of catheters to provide quality health care and avoid unnecessary catheterization [1]. The aim of this study was to audit the appropriateness of usage of urinary catheters by the healthcare staff and, their technique of insertion and indications for urinary catheterization. What are your research's implications towards theory, practice, or policy? Hand hygiene is an important element of patient care. If done properly it can prevent several hospital-acquired infections. Heathcare facilities must audit their staff on regular basis and provide refreshers and promote the use of proper hand hygiene before urinary catheterization.

The effectiveness of high-flow nasal cannula and standard non-rebreathing mask for oxygen therapy in moderate category COVID-19 pneumonia: Randomised controlled trial.

Background: COVID-19 caused by the highly infectious severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) infection is a matter of concern and has led to severe health problems all over the world. Oxygen therapy is the mainstay for the management of patients suffering from various stages of the disease. Objectives: To compare the effectiveness of high-flow nasal cannula (HFNC) and standard non-rebreathing mask (NRBM) as oxygen delivery devices in moderate cases of COVID-19 pneumonia. Methods: A single-centre, open-label, randomised controlled trial was conducted between February 2021 and April 2021. All the enrolled patients (N=120) were randomly allocated into two groups according to the oxygen delivery device used. Group 1 (n=60) received HFNC and group 2 (n=60) received NRBM as the initial oxygen delivery device, to maintain a target saturation ≥96% in both groups. The progression-free survival without escalation of respiratory support, partial pressure of arterial oxygen (PaO2 ), a ratio of partial pressure of arterial oxygen to fractional inspiratory oxygen concentration (PaO2 /FiO2 ), respiratory rate, heart rate, blood pressure, number of patients requiring non-invasive ventilation or endotracheal intubation, time for de-escalation of oxygen therapy to lower FiO2 device, time to progression to severe disease, survival at day 28, and patient satisfaction level were compared between the two groups. Results: Demographic, clinical variables and treatment received were comparable in the two groups. In the HFNC group, 90% of patients had successful outcomes with the initial oxygen therapy device used as compared with 56.6% in the NRBM group (p<0.001; odds ratio (OR) 0.145; 95% confidence interval (CI) 0.054 - 0.389). Using HFNC also resulted in improved oxygenation (PaO2 /FiO2 ) (p<0.001), better patient satisfaction (p<0.001), and a shorter time for de-escalation of oxygen therapy to a lower FiO2 device (p<0.001). The 28-day survival was higher in the HFNC group, but the difference was statistically insignificant (p=0.468). Conclusion: HFNC is a reliable oxygen therapy modality for moderate category COVID-19 pneumonia and results in a higher success rate of oxygen therapy, better oxygenation, and a greater patient satisfaction level as compared with a NRBM.

Molecular and phenotypic characterization of two cryptic species of the predatory genus Mononchoides Rahm, 1928 (Rhabditida: Diplogastridae) and their congeneric affinities.

Based on morphometric, morphological and molecular characterization using partial small subunit 18S ribosomal DNA (rDNA) and the D2/D3 domain of large subunit 28S rDNA, we described a new species Mononchoides kanzakii collected from manure, and the known species Mononchoides composticola Steel, Moens, Scholaert, Boshoff, Houthoofd and Bert, 2011, isolated from the dung beetle Oniticellus cinctus (Fabricius, 1775). Phylogenetic trees based on the evolutionary model (GTR + I + G) were inferred by Bayesian inference algorithms. Mononchoides kanzakii sp. n. is characterized by 28-32 longitudinal ridges, discontinuous at level of stoma; amphidial apertures inconspicuous; metastegostom armed with thorn-shaped dorsal tooth; a flattened, claw-like right subventral tooth, and left subventral denticulate ridge with 12-14 fine denticles delimited by a group of five denticles in females vs. triangular, flattened right subventral tooth, 5-8 prominent denticles at left subventral sector in males; cloacal lips with a distinct rim; and gubernaculum with cuticularized, proximal and distal extensions of equal length, each constituting half of the length of the wider part of gubernaculum.

It's not just about recruitment: An exploratory look at tobacco education sessions to increase participation into smoking cessation programs among American Indians.

American Indians (AI) have the highest smoking rates and lowest quit rates of any racial/ethnic group in the U.S. Researchers and community members from the American Indian Health Research and Education Alliance (AIHREA) created and evaluated a culturally-tailored smoking cessation program, All Nations Breath of Life (ANBL) as a recruitment tool for smoking cessation programs among AI. To increase enrollment in ANBL, AI smokers were approached at cultural events and asked to attend a 30-minute educational session (in-person, n= 179; tele-video, n=97). Tele-video (30%) and in-person (9%) session participants were recruited into ANBL. Pre- and post-tests showed participants in both sessions demonstrated increased motivation and confidence to quit smoking but significant differences were present in both sessions (p < 0.0001). Results indicate that theoretically guided and culturally tailored education sessions are viable approaches to educate and recruit underserved populations into programs that promote smoking cessation.

Association of single nucleotide polymorphisms in microRNAs with susceptibility to hepatitis B virus infection and HBV-related liver complications: A study in a Saudi Arabian population.

The aim of this study was to evaluate the association of 10 SNPs in different microRNAs (miRNAs) with susceptibility to hepatitis B virus (HBV) infection, HBV clearance, persistence of chronic HBV infection, and progression to liver cirrhosis and hepatocellular carcinoma (HCC). Patients were categorized into the following groups: inactive HBV carrier, active HBV carrier, HBV-cleared subject and cirrhosis+HCC. Samples were analysed for 10 SNPs in microRNAs using either PCR-based genotyping or the TaqMan assay. We found that rs1358379 was associated with susceptibility to HBV infection, HBV clearance, persistent chronic HBV infection and liver cirrhosis+HCC. In addition, we found that rs2292832 and rs11614913 were associated with risk of HBV infection, viral clearance and cirrhosis+HCC, whereas rs2910164 was associated with proneness to HBV infection, and ability to clear the virus. There was evidence of associations between rs6505162 and HBV clearance and the development of liver disease, whereas a single association was found between rs2289030 and HBV clearance. Similarly, rs7372209 and rs4919510 were specifically associated with the development of HBV-induced liver complications. SNPs in miRNAs affect the susceptibility, clearance and progression of HBV infection in Saudi Arabian patients. We found, using Gene Ontology or pathway analyses, that these genes may contribute to the pathophysiology of HBV infection and related liver complications. However, differences in the association of examined SNPs with various clinical stages indicate variations in the respective functional roles of these polymorphisms and their miRNAs, and thus, further investigation to fully explore their therapeutic potential is warranted.

Antibody Prevalence and Risk Factors for Toxoplasma gondii Infection in Women from Multan, Pakistan.

Toxoplasma gondii infections are prevalent in humans and warm-blooded animals. Maternal infections during pregnancy may have devastating consequences for transplacentally infected neonates. This study was conducted to examine the seroprevalence of antibodies to T. gondii in pregnant women of childbearing age and determine risk factors associated with pregnancy history, pet ownership, social and cultural factors at Nishtar Hospital, Multan. Samples were collected from 403 women and examined using a commercially available enzyme-linked immunosorbent assay (ELISA). The overall prevalence of antibodies to T. gondii was 17.6% (71) in the 403 samples collected from women. Antibodies to T. gondii were present in 19.4% (45) of 232 pregnant women and 15.2% (26) of the samples from 171 non-pregnant women. This study identified miscarriage history, pet ownership, type of residence, marital status, source of drinking water and eating habits as significant (P < 0.05) risk factors associated with the presence of antibodies to T. gondii infection. Seroprevalence was not significantly different (P > 0.05) in women from different ethnic groups based upon lifestyle and culture.

Fecal occult blood test use by Kansas medicare beneficiaries.

PURPOSE: The aim of this study was to examine utilization of and trends in fecal occult blood testing (FOBT) among beneficiaries since Medicare began FOBT coverage on January 1, 1998. METHODS: We identified Kansas Medicare beneficiaries ages 65-79. Using Medicare claims, we determined which beneficiaries received FOBT during 1998-1999. We examined demographic variables associated with FOBT and rate changes over time. We linked beneficiaries to primary care practices (PCPs) and examined FOBT variations among 483 PCPs. RESULTS: FOBT use remained unchanged during the study period. Of the 215,322 beneficiaries, 11% received at least one FOBT in 1998 and 11% in 1999; 18% had at least one test during the 2-year period, but only 4% had a test during both years. Caucasians and females had the highest FOBT rates. Although FOBT rates among PCPs ranged from 0 to 71%, only 19% of the practice rates exceeded 10%. CONCLUSIONS: Few beneficiaries obtain annual FOBT and little change in rates has occurred since Medicare reimbursement began. Although FOBT rates vary widely between PCPs, most either do not provide FOBT or do not bill Medicare for FOBT. The FOBT claims rate is much lower than reported in patient surveys and may indicate that Medicare should reexamine its reimbursement policy.

Comparative salt tolerance of amphidiploid and diploid Brassica species.

Salt (NaCl) tolerance of three amphidiploid Brassica species, B. napus (AC genome), B. carinata (BC genome), and B. juncea (AB genome), and their putative diploid relatives, B. campestris (A genome), B. oleracea (C genome) and B. nigra (B genome) was examined under glasshouse conditions. The plants were grown in sand culture throughout the study period (51 days). Twenty-three day old plants of all six species were subjected for 28 days to control (0 mol m(-3) NaCl), 100 and 200 mol m(-3) NaCl in Hoagland nutrient solution. Under saline conditions, growth of the three amphidiploid species was significantly greater in shoot and root weights, and seed yield was greater than that of the diploids. The amphidiploids accumulated lower Na(+) but higher K(+) in their shoots and roots, the K/Na ratio therefore being considerably higher than those of the diploids. Ca(2+) accumulation was similar in the diploids and amphidiploids, and the Cl(-) accumulation pattern was not consistent in the different species. The occurrence of high salt tolerance of amphidiploids, and their enhanced K/Na discrimination with respect to their diploid relatives, suggests that salt tolerance has been obtained from A and C genomes and the latter trait from all three genomes (A, B, and C).