Patients' knowledge, attitude, and practices toward unused medications in Qatar: A cross-sectional survey.

Background: Global evidence has linked unused medications and their inappropriate disposal to adverse health, economic, environmental, and ethical impacts. However, such evidence is scarce in Qatar. This study explored patients' knowledge and attitude toward unused medications and their practices toward medication supply and disposal (KAP) in Qatar.Materials and methods. Study design: A cross-sectional survey using a pretested questionnaire was performed between February 2020 and October 2020. Descriptive statistics, Man Whitney U, and Kruskal-Wallis Rank-Sum tests were applied. The Chi square test assessed the association between socio-demographic characteristics and KAP scores. Characteristics that were found significantly associated with KAP (i.e., p-values <0.05) were further included as predictor variables in the multiple linear regression model. Results: All items pertaining to patients' knowledge were found to be good (mean score > 3), except for "awareness of unwanted medication return policy" (mean score < 3), i.e., the lowest level of patient agreement (31 %) (median (M) = 3, Interquartile Range (IQR) = 3). Their attitude was generally good (mean score > 3). Conversely, their practice toward medication supply was poor (mean score < 3). Possible future use was the most reported reason (79 %) for keeping medications at home, and home trash was the most widely disposing place of unused ones (76 %). Knowledge was significantly higher among non-laborers and other occupations than among patients with no work (p < 0.001) and (p = 0.005), respectively. The attitude was significantly lower among patients with healthcare providers (HCPs) in their household than among those without (p = 0.001). Practices were also significantly lower among those aged 40-49 years and those with HCPs in their household than those aged 18-29 years (p = 0.012) and those without HCPs, (p < 0.001), respectively. Conclusions: Overall, patients' knowledge and attitude toward unused medications seem good, while their practices toward medication supply and disposal are bad. To mitigate the health, economic, and environmental impacts of unused medications, interventions including rationalizing drug supply, use, disposal, prescribing, manufacturing, and promotion are recommended.

Adductor Canal Block Versus Local Infiltration Analgesia for Postoperative Pain Management in Total Knee Arthroplasty.

BACKGROUND: Total knee arthroplasty (TKA) is a common surgical procedure for patients with knee osteoarthritis, often associated with postoperative pain. Effective pain management strategies are essential for improving patient outcomes and satisfaction. This study aimed to compare the efficacy of two analgesic modalities, local infiltration analgesia (LIA) and adductor canal block (ACB), in providing postoperative pain relief for patients undergoing TKA. METHODS: This prospective randomized comparative study included 60 patients undergoing TKA for knee osteoarthritis under subarachnoid block (spinal anaesthesia). Patients were divided into two groups: LIA group (local wound infiltration with periarticular injection of bupivacaine 0.125% + dexmedetomidine 1 mcg/kg) and ACB group (ACB with bupivacaine 0.125% + 1 mcg/kg dexmedetomidine). Pain relief was assessed using the Numerical Rating Scale (NRS) score, time to first rescue analgesic requirement (NRS > 3), and total amount of analgesic needed in the first 24 hours post-surgery. RESULTS: The time to first perception of pain with NRS > 3 was 11.30±0.8 hours in the ACB group and 9.40 ± 1.1 hours in the LIA group, with a statistically significant difference (p < 0.001). Additionally, the total number of rescue analgesic doses given in the first 24 hours post-operatively differed significantly between the two groups (p = 0.046). CONCLUSION: The study concludes that ACB is an effective postoperative analgesic modality, superior to local infiltration analgesia, for patients undergoing TKA.

Outcome of a Simple Novel Technique to Reduce Soft Tissue Complications in Open Tendoachilles Injury: A Series of 20 Patients.

Introduction: Open tendoachilles injuries are rare and associated with significant soft tissues complications. The objective of the present study was to assess the clinical outcome and safety of a simple and minimally invasive technique, with a goal to assess if it may help minimise flap and wound related complications in open tendoachilles injuries. Materials and methods: This prospective study of four years duration included 20 patients with open tendoachilles injuries managed with a simple minimally invasive tunnel technique. The primary outcome variable was occurrence of a major soft tissue complication. The secondary outcome variables included functional outcome measured using AOFAS Ankle hind foot score, re-rupture of tendoachilles and need for revision surgery. Results: None of the patients in the present series developed a serious soft tissue complication. Based upon the AOFAS hind foot scoring system, good to excellent outcome was achieved in 19 (95%) patients. All the patients were able to perform tip toe walking at six months post-surgery. None of the patients had a re-rupture of the tendoachilles and no patient needed a revision surgery. The complications encountered include thickening of the tendon at the repair site (15%), superficial wound infection (5%), stitch granuloma (5%) and hypertrophic scar (5%). Conclusion: This technique seems to be promising in reducing the soft tissue complications associated with the surgical management of open tendoachilles injuries. Most patients had a good final clinical outcome. The technique is safe, simple and reproducible. However, further randomised control studies with a larger sample size assessing the technique are recommended.

Discovery of SDS-347 as a specific peptide competitive inhibitor of G9a with promising anti-cancer potential.

BACKGROUND: G9a is a histone H3K9 methyltransferase enzyme found highly upregulated in many cancers. H3 binds to the rigid I-SET domain and the cofactor, S-adenosyl methionine, binds to the flexible post-SET domain of G9a. Inhibition of G9a is known to inhibit the growth of cancer cell-lines. METHODS: Recombinant G9a and H3 were used to develop radioisotope-based inhibitor screening assay. The identified inhibitor was evaluated for isoform selectivity. The mode of enzymatic inhibition was studied by enzymatic assays and bioinformatics. Anti-proliferative activity of the inhibitor was studied in cancer cell lines by utilizing MTT assay. The mechanism of cell death was studied by western blotting and microscopy. RESULTS: We developed a robust G9a inhibitor screening assay that led to the discovery of SDS-347 as a potent G9a inhibitor with IC50 of 3.06 µM. It was shown to reduce the levels of H3K9me2 in cell-based assay. The inhibitor was found to be peptide competitive and highly specific as it did not show any significant inhibition of other histone methyltransferases and DNA methyltransferase. Docking studies showed that SDS-347 could form direct bonding interaction with Asp1088 of the peptide-binding site. SDS-347 showed anti-proliferative effect against various cancer cell lines especially the K562 cells. Our data suggested that SDS-347 mediated antiproliferative action via ROS generation, induction of autophagy and apoptosis. CONCLUSION: Overall, the findings of the current study include development of a new G9a inhibitor screening assay and identification of SDS-347, as a novel, peptide competitive and highly specific G9a inhibitor with promising anticancer potential.

Enhancing the biochemical growth of Haematococcus pluvialis by mitigation of broad-spectrum light stress in wastewater cultures.

In this study, the microalgae Haematococcus pluvialis were cultivated in wastewater inoculated into low-density polypropylene plastic air pillows (LDPE-PAPs) under a light stress. The cells were irradiated to different light stresses using white LED lights (WLs) as the control, and broad-spectrum lights (BLs) as a test for the period of 32 days. It was observed that the inoculum (70 × 102 mL-1 cells) of H. pluvialis algal cells increased almost 30 and 40 times in WL and BL, respectively, at day 32 coherent to its biomass productivity. Higher lipid concentration of up to 36.85 µg mL-1 was observed in BL irradiated cells compared to 13.215 µg L-1 dry weight of biomass in WL. The chlorophyll 'a' content was 2.6 times greater in BL (3.46 µg mL-1) compared to that in WL (1.32 µg mL-1) with total carotenoids being about 1.5 times greater in BL compared to WL on day 32. The yield of red pigment 'Astaxanthin' was about 27% greater in BL than in WL. The presence, of different carotenoids including astaxanthin was also confirmed by HPLC, whereas fatty acid methyl esters (FAMEs) were confirmed by GC-MS. This study further confirmed that wastewater alongwith with light stress is suitable for the biochemical growth of H. pluvialis with good biomass yield as well as carotenoid accumulation. Additionally there was 46% reduction in chemical oxygen demand (COD) in a far more efficient manner when cultured in recycled LDPE-PAP. Such type of cultivation of H. pluvialis made the overall process economical and suitable for upscaling to produce value-added products such as lipids, pigments, biomass, and biofuel for commercial applications.

Fit-cardiopulmonary resuscitation approach in public mass cardiopulmonary resuscitation teaching: A randomized control trial.

OBJECTIVES: To improve public awareness and the rate of bystander cardiopulmonary resuscitation (CPR), a novel and exciting approach called fit-CPR that incorporates mass CPR with high-intensity physical activity into the beat of locally favoured music was proposed. This study was conducted to measure the effectiveness of fit-CPR compared to the standard classroom method (CCM). METHODS: Between 30th August to 29th November 2018, 129 participants from Syiah Kuala University, Banda Aceh, Indonesia, were randomized to learn CPR, either through fit-CPR or CCM protocol. All participants underwent pre, post, and 6-month retention tests. Each test had a 10-item questionnaire with CPR performance on a manikin that was assessed using a validated checklist. RESULTS: Sixty-one (47.3%) participants completed the fit-CPR while 68 (52.7%) completed the CCM. There was a significant improvement in knowledge, performance, and quality of CPR from pre, post, and 6-month retention tests (p<0.01) in both groups. On high-quality CPR, the fit-CPR and CCM groups obtained an increased score of 285.0% and 151%, respectively, p=0.014 between pre and immediate post-test. Knowledge scores between fit-CPR and CCM groups showed an increase of 79.5% and 111.2%, respectively, p=0.002. Fit-CPR was completed between 52.5-57.5 minutes, while CCM took 75 minutes. CONCLUSION: The fit-CPR demonstrated a comparable outcome to standard CPR when teaching to the mass public with less time spent.

Outcome of a Simple Novel Technique to Reduce Soft Tissue Complications in Open Tendoachilles Injury: A Series of 20 Patients

@#Introduction: Open tendoachilles injuries are rare and associated with significant soft tissues complications. The objective of the present study was to assess the clinical outcome and safety of a simple and minimally invasive technique, with a goal to assess if it may help minimise flap and wound related complications in open tendoachilles injuries. Materials and methods: This prospective study of four years duration included 20 patients with open tendoachilles injuries managed with a simple minimally invasive tunnel technique. The primary outcome variable was occurrence of a major soft tissue complication. The secondary outcome variables included functional outcome measured using AOFAS Ankle hind foot score, re-rupture of tendoachilles and need for revision surgery. Results: None of the patients in the present series developed a serious soft tissue complication. Based upon the AOFAS hind foot scoring system, good to excellent outcome was achieved in 19 (95%) patients. All the patients were able to perform tip toe walking at six months post-surgery. None of the patients had a re-rupture of the tendoachilles and no patient needed a revision surgery. The complications encountered include thickening of the tendon at the repair site (15%), superficial wound infection (5%), stitch granuloma (5%) and hypertrophic scar (5%). Conclusion: This technique seems to be promising in reducing the soft tissue complications associated with the surgical management of open tendoachilles injuries. Most patients had a good final clinical outcome. The technique is safe, simple and reproducible. However, further randomised control studies with a larger sample size assessing the technique are recommended.

The study of border zone formation in ischemic heart using electro-chemical coupled computational model.

Myocardial infarction (MI) is the most common cause of a heart failure, which occurs due to myocardial ischemia leading to left ventricular (LV) remodeling. LV remodeling particularly occurs at the ischemic area and the region surrounds it, known as the border zone. The role of the border zone in initiating LV remodeling process urges the investigation on the correlation between early border zone changes and remodeling outcome. Thus, this study aims to simulate a preliminary conceptual work of the border zone formation and evolution during onset of MI and its effect towards early LV remodeling processes by incorporating the oxygen concentration effect on the electrophysiology of an idealized three-dimensional LV through electro-chemical coupled mathematical model. The simulation result shows that the region of border zone, represented by the distribution of electrical conductivities, keeps expanding over time. Based on this result, the border zone is also proposed to consist of three sub-regions, namely mildly, moderately, and seriously impaired conductivity regions, which each region categorized depending on its electrical conductivities. This division could be used as a biomarker for classification of reversible and irreversible myocardial injury and will help to identify the different risks for the survival of patient. Larger ischemic size and complete occlusion of the coronary artery can be associated with an increased risk of developing irreversible injury, in particular if the reperfusion treatment is delayed. Increased irreversible injury area can be related with cardiovascular events and will further deteriorate the LV function over time.

3'-(4-(Benzyloxy)phenyl)-1'-phenyl-5-(heteroaryl/aryl)-3,4-dihydro-1'H,2H-[3,4'-bipyrazole]-2-carboxamides as EGFR kinase inhibitors: Synthesis, anticancer evaluation, and molecular docking studies.

Pyrazoline-linked carboxamide derivatives were designed, synthesized, and evaluated for potential epidermal growth factor receptor (EGFR) kinase inhibition, anticancer activity, and apoptotic and cardiomyopathy toxicity. Compounds 6m and 6n inhibit EGFR kinase at a concentration of 6.5 ± 2.91 and 3.65 ± 0.54 µM, respectively. Some of these compounds showed effects on proliferation, which were also then evaluated against four different human cancer cell lines, that is, MCF-7 (breast cancer), A549 (non-small-cell lung tumor), HCT-116 (colon cancer), and SiHa cells (cancerous tissues of the cervix uteri). The results showed that certain synthetic compounds showed significant inhibitor activity; compounds 6m and 6n were more cytotoxic than doxorubicin against A549 cancer cells, with IC50 values of 10.3 ± 1.07 and 4.6 ± 0.57 µM, respectively. Additionally, compounds 6m and 6n induced apoptosis in A549 cancer cells, as evidenced by 4',6-diamidino-2-phenylindole (DAPI) staining and phase-contrast microscopy. Potency to induce apoptosis by compound 6n was further confirmed by fluorescence-activated cell sorting using Annexin V-FITC and propidium iodide labeling. Compound 6n showed normal cardiomyocytes with no marked sign of pyknotic nuclei in cardiomyopathy and also normal histological appearance of the renal cortex when compared with that of control. Results of molecular docking studies suggested that compounds 6m and 6n can bind to the hinge region of the adenosine triphosphate-binding site of EGFR kinase, like the standard drug erlotinib. Therefore, the present study suggests that compounds 6m and 6n have potent in vitro antitumor activities against the human non-small-cell lung tumor cell line A549, which can be further explored in other cancer cell lines and in animal studies.

Differentiation of human neuroblastoma cell line IMR-32 by sildenafil and its newly discovered analogue IS00384.

Sildenafil, a phosphodiesterase-5 inhibitor is FDA approved drug against erectile dysfunction. It is currently undergoing many clinical trials, alone or in combinations against different diseases. Treatment of neural progenitor cells with sildenafil is known to regulate their basal cGMP levels and enhance neurogenesis and differentiation. cGMP as well as cAMP are known to play a central role in the maintenance, repair and remodelling of the nervous system. In the present study, we report the neurodifferentiation property of sildenafil in neuroblastoma cancer cell line IMR-32. Sildenafil was found to induce the formation of neurite outgrowths that were found expressing neuronal markers, such as NeuN, NF-H and ßIII tubulin. IS00384, a recently discovered PDE5 inhibitor by our laboratory, was also found to induce neurodifferentiation of IMR-32 cells. The effect of IS00384 on differentiation was even more profound than sildenafil. Both the compounds were found to elevate and activate the Guanine nucleotide exchange factor C3G, which is a regulator of differentiation in IMR-32 cells. They were also found to elevate the levels of cGMP and activate the AMPK-ACC and PI3K-Akt signalling pathways. These pathways are known to play important role in cytoskeletal rearrangements necessary for differentiation. This study highlights the role of phosphodiesterases-5 in neurodifferentiation and use of sildenafil and IS00384 as small molecule tools to study the process of cellular differentiation.

Synthesis of Dihydropyrimidinones (DHPMs) and Hexahydro Xanthene Catalyzed by 1,4-Diazabicyclo [2.2.2] Octane Triflate Under Solvent-Free Condition.

OBJECTIVE: DABCO salts were evaluated as catalysts for the Biginelli reaction between 4- methoxybenzaldehyde, urea and ethyl acetoacetate under solvent-free conditions. 1,4-Diazabicyclo [2.2.2] octane triflate was found to be a simple, inexpensive, highly efficient catalyst for Biginelli reaction for a variety aromatic aldehyde with urea and ethyl acetoacetate at 80°C afforded corresponding 3,4-dihydropyrimidinones in 50-99% yields after 30-120 minutes. 1,3-Cyclohexadione was used in place of ethyl acetoacetate in the absence of urea this methodology is giving hexahydro xanthene derivatives in good to excellent yields after 3-4 hours. METHODS: DABCO salt 4 (5 mol%), 4-methoxybenzaldehyde (0.73 mmol) and urea (0.73 mmol) were stirred for 10 minutes at 80°C, then ethyl acetoacetate (1.5 equiv.) was added and reaction mixture was stirred at 80°C for specified time. The resulting solution was stirred continuously and progress of the reaction was followed by TLC. The crude reaction mixture was purified by flash column chromatography on silica gel (hexane/ethyl acetate (1:2)) to give pure desired product. RESULTS: Reaction conditions of the Biginelli reaction were optimized using 4-methoxybenzaldehyde (0.73 mmol), urea (0.73 mmol), and ethyl acetoacetate (5 equiv.) as model substrates catalyzed by 1,4-Diazabicyclo [2.2.2] octane triflate (5 mol%) in a different solvents, screening of different catalysts and different temperatures. Neat condition was found to be the best for the Biginelli condensation and corresponding 3,4- dihydropyrimidinones was obtained in good to excellent yields. When the reaction was carried out with benzaldehyde derivatives and cyclohexane-1,3-dione in place of ethyl acetoacetate in the absence of urea, solely corresponding hexahydro xanthene derivatives were obtained in 61-91% yields. CONCLUSION: In conclusion, we have applied salts of 1,4-Diaza-bicyclo [2.2.2] octane as catalysts in the Biginelli condensation and corresponding 3,4-dihydropyrimidinones were obtained in 50- 99% yields under solvent free conditions. This methodology is having advantages like simple work-up; low loading of catalyst and reaction was performed at moderate temperature under solvent-free conditions.

A cross sectional study on the acceptance of pre-hospital continuous positive airway pressure ventilation among ambulance paramedic in an urban emergency medical service system in a developing country.

OBJECTIVES: To assess the acceptance among the developing country urban paramedics towards pre-hospital continuous positive airway pressure (CPAP) ventilation. METHODS: A cross-sectional prospective study was conducted among the ambulance paramedics working at the pre-hospital care unit of the Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia from March 2012 to August 2012 on ambulance paramedics. Questionnaires were used to assess their experience, knowledge, and perception, while their competencies were assessed using an Objective Structured Clinical Examination by 2 independent-calibrated raters on the use of the Boussignac CPAP system. RESULTS: Twenty-six ambulance paramedics qualified for this study with an average work experience of 5.59+/-3.53 years. A total of 76.9% had no formal training for CPAP during their study years. Knowledge of CPAP apparatus-arrangement sequence scored as 88.5% correct, while 96.2% scored `Good` to `Very-good` in the ability to diagnose conditions that warrant its use. A total of 76.9% were confident to monitor patients on CPAP, and 61.5% in applying the device. However, only 53.8% were confident to start the CPAP, and 38.5% to troubleshoot if any problem arose. For perceptions, 96.2% felt it was easy to learn CPAP, while 88.5% felt that paramedics could use it without supervision, and 80.8% felt that it should not be confined to the Emergency Department setting. A total of 96.1% were competent in CPAP application. CONCLUSION: Developing country urban ambulance paramedics possessed adequate knowledge, positive attitudes, and demonstrated good CPAP application skills. However, lack of confidence towards decision to initiate and troubleshoot of potential complications were the main obstacles hindering its use.

Improving door-to-doctor and door-to-analgesia time in a tertiary teaching hospital emergency department through a novel 'Red Box' system.

OBJECTIVE: To evaluate the effectiveness of a new patient flow system, `The Red Box` on the quality of patient care in respect of the time taken for the care to be delivered to the patient. METHODS: A pre-post study was conducted looking at the door-to-doctor (DTD) and door-to-analgesia (DTA) times for cases presenting to the Emergency Department (ED) of a tertiary teaching hospital `The National University of Malaysia Medical Center` between the periods of July and September 2005 against July and September 2008. Demographic data, ED presentation time, time seen by first doctor, and time first analgesia given were collected in both periods and analyzed. RESULTS: A total of 1,000 cases were enrolled. Group A (pre-Red Box) and group B (post-Red Box) comprised 500 cases each. The mean DTD time for group A was 29 minutes (SD +/- 3 minutes) and for group B was 3 minutes (SD +/- 1 minute), with a 98.8% reduction (p<0.001). For DTA time, group A recorded a mean of 46 minutes (SD +/- 3 minutes), and group B recorded a mean of 9 minutes (SD +/- 2 minutes), an 80.4% reduction (p<0.001). CONCLUSION: The implementation of a red box system improved the quality of emergency patient care in the ED of a tertiary teaching hospital as evidenced by significant reductions in DTD and DTA time.

Single-port access total laparoscopic hysterectomy for stage 1A1 cancer of the cervix.

We describe the first case of single-port access total laparoscopic hysterectomy with intracorporeal suturing of the vault performed in Singapore. A 40-year-old woman with microinvasive squamous cell carcinoma of the cervix successfully underwent single-port access total laparoscopic hysterectomy. Unique articulated and multifunction laparoscopic instruments were used to complete the surgery in 118 minutes, with no complications. The patient had minimal pain postoperatively and recovered uneventfully within two weeks. This case illustrates the benefits of single-port access laparoscopic surgery in well-selected cases.

Genotoxic potential of copper oxide nanoparticles in human lung epithelial cells.

Copper oxide nanoparticles (CuO NPs) are increasingly used in various applications. Recent studies suggest that oxidative stress may be the cause of the cytotoxicity of CuO NPs in mammalian cells. However, little is known about the genotoxicity of CuO NPs following exposure to human cells. This study was undertaken to investigate CuO NPs induced genotoxic response through p53 pathway in human pulmonary epithelial cells (A549). In addition, cytotoxicity and oxidative stress markers were also assessed. Results showed that cell viability was reduced by CuO NPs and degree of reduction was dose dependent. CuO NPs were also found to induce oxidative stress in dose-dependent manner indicated by depletion of glutathione and induction of lipid peroxidation, catalase and superoxide dismutase. The expression of Hsp70, the first tier biomarker of cellular damage was induced by CuO NPs. Further, CuO NPs up-regulated the cell cycle checkpoint protein p53 and DNA damage repair proteins Rad51 and MSH2 expression. These results demonstrate that CuO NPs possess a genotoxic potential in A549 cells which may be mediated through oxidative stress. Our short-term exposure study of high level induction of genotoxic response of CuO NPs will need to be further investigated to determine whether long-term exposure consequences may exist for CuO NPs application.

Chronopharmaceutics: a promising drug delivery finding of the last two decades.

Pulsatile drug delivery system capable of releasing the drug after a predetermined lag period in pulsed or controlled release manner recently has drawn the attention of both academic and industrial research. Depending on the effective therapeutic application of the drug, a variety of design strategies have been formulated in the pursuit of pulsatile release. Circadian (24 hr cycle) dependency of various physiological and pathological functions is well established, thus, it becomes imperative to develop a drug delivery system to achieve release of drug at specific site and time. Such systems are advantageous for drugs which have an extensive first pass metabolism, biological tolerance, needs targeting of locally absorbed / active drug to a specific site in intestine and are useful for the therapy for chronopharmacological needs. This manuscript portrays the important patents related to chronomodulated release system such as system with eroding, rupturing or soluble barrier coatings. In addition, recently developed chronotherapeutic dosage forms including tablets, capsules, pellets, beads implants, osmotic pump, liposome, thermoresponsive, inflammation stimuli sensitive, electrical stimuli sensitive, ultrasound stimuli responsive, magnetic stimuli responsive etc., are also conferred.

The requirement for cellular transportin 3 (TNPO3 or TRN-SR2) during infection maps to human immunodeficiency virus type 1 capsid and not integrase.

Recent genome-wide screens have highlighted an important role for transportin 3 in human immunodeficiency virus type 1 (HIV-1) infection and preintegration complex (PIC) nuclear import. Moreover, HIV-1 integrase interacted with recombinant transportin 3 protein under conditions whereby Moloney murine leukemia virus (MLV) integrase failed to do so, suggesting that integrase-transportin 3 interactions might underscore active retroviral PIC nuclear import. Here we correlate infectivity defects in transportin 3 knockdown cells with in vitro protein binding affinities for an expanded set of retroviruses that include simian immunodeficiency virus (SIV), bovine immunodeficiency virus (BIV), equine infectious anemia virus (EIAV), feline immunodeficiency virus (FIV), and Rous sarcoma virus (RSV) to critically address the role of integrase-transportin 3 interactions in viral infection. Lentiviruses, with the exception of FIV, display a requirement for transportin 3 in comparison to MLV and RSV, yielding an infection-based dependency ranking of SIV > HIV-1 > BIV and EIAV > MLV, RSV, and FIV. In vitro pulldown and surface plasmon resonance assays, in contrast, define a notably different integrase-transportin 3 binding hierarchy: FIV, HIV-1, and BIV > SIV and MLV > EIAV. Our results therefore fail to support a critical role for integrase binding in dictating transportin 3 dependency during retrovirus infection. In addition to integrase, capsid has been highlighted as a retroviral nuclear import determinant. Accordingly, MLV/HIV-1 chimera viruses pinpoint the genetic determinant of sensitization to transportin 3 knockdown to the HIV-1 capsid protein. We therefore conclude that capsid, not integrase, is the dominant viral factor that dictates transportin 3 dependency during HIV-1 infection.

Biochemical and virological analysis of the 18-residue C-terminal tail of HIV-1 integrase.

BACKGROUND: The 18 residue tail abutting the SH3 fold that comprises the heart of the C-terminal domain is the only part of HIV-1 integrase yet to be visualized by structural biology. To ascertain the role of the tail region in integrase function and HIV-1 replication, a set of deletion mutants that successively lacked three amino acids was constructed and analyzed in a variety of biochemical and virus infection assays. HIV-1/2 chimers, which harbored the analogous 23-mer HIV-2 tail in place of the HIV-1 sequence, were also studied. Because integrase mutations can affect steps in the replication cycle other than integration, defective mutant viruses were tested for integrase protein content and reverse transcription in addition to integration. The F185K core domain mutation, which increases integrase protein solubility, was furthermore analyzed in a subset of mutants. RESULTS: Purified proteins were assessed for in vitro levels of 3' processing and DNA strand transfer activities whereas HIV-1 infectivity was measured using luciferase reporter viruses. Deletions lacking up to 9 amino acids (1-285, 1-282, and 1-279) displayed near wild-type activities in vitro and during infection. Further deletion yielded two viruses, HIV-1(1-276) and HIV-1(1-273), that displayed approximately two and 5-fold infectivity defects, respectively, due to reduced integrase function. Deletion mutant HIV-1(1-270) and the HIV-1/2 chimera were non-infectious and displayed approximately 3 to 4-fold reverse transcription in addition to severe integration defects. Removal of four additional residues, which encompassed the C-terminal beta strand of the SH3 fold, further compromised integrase incorporation into virions and reverse transcription. CONCLUSION: HIV-1(1-270), HIV-1(1-266), and the HIV-1/2 chimera were typed as class II mutant viruses due to their pleiotropic replication defects. We speculate that residues 271-273 might play a role in mediating the known integrase-reverse transcriptase interaction, as their removal unveiled a reverse transcription defect. The F185K mutation reduced the in vitro activities of 1-279 and 1-276 integrases by about 25%. Mutant proteins 1-279/F185K and 1-276/F185K are therefore highlighted as potential structural biology candidates, whereas further deleted tail variants (1-273/F185K or 1-270/F185K) are less desirable due to marginal or undetectable levels of integrase function.

Prevalence and pattern of congenital heart disease in school children of eastern Uttar Pradesh.

BACKGROUND AND AIM: The prevalence of congenital heart disease (CHD) is not known in our country. The aim of present study was to find out the prevalence of CHD in school children of eastern Uttar Pradesh. METHOD: A team consisting of a cardiologist, physicians and junior residents visited schools in the area. All the children were examined for presence of cardiac murmur or history of heart disease or any intervention. Those with murmurs or previous history of heart disease were called to the Medical College Hospital for evaluation by ECG, chest X-ray and echocardiography for confirmation of the lesion. RESULTS: Out of 118,212 children examined, 142 were found to have CHD. The prevalence was 1.3 per 1000 children and the commonest lesions were ventricular and atrial septal defects, aortic stenosis with or without regurgitation, and pulmonary stenosis. CONCLUSION: CHD prevalence is 1.3 per 1000 school children that is nearly two and a half times more than that of RHD. Knowing it is important for development of facilities for CHD care in our setup.