Virtual Screening of FDA-Approved Drugs against LasR of Pseudomonas aeruginosa for Antibiofilm Potential.

Pseudomonas aeruginosa is a Gram-negative pathogenic bacterium that is present commonly in soil and water and is responsible for causing septic shock, pneumonia, urinary tract and gastrointestinal infections, etc. The multi-drug resistance (MDR) phenomenon has increased dramatically in past years and is now considered a major threat globally, so there is an urgent need to develop new strategies to overcome drug resistance by P. aeruginosa. In P. aeruginosa, a major factor of drug resistance is associated to the formation of biofilms by the LasR enzyme, which regulates quorum sensing and has been reported as a new therapeutic target for designing novel antibacterial molecules. In this study, virtual screening and molecular docking were performed against the ligand binding domain (LBD) of LasR by employing a pharmacophore hypothesis for the screening of 2373 FDA-approved compounds to filter top-scoring hit compounds. Six inhibitors out of 2373 compounds were found to have binding affinities close to that of known LasR inhibitors. The binding modes of these compounds to the binding site in LasR-LBD were analyzed to identify the key interactions that contribute to the inhibition of LasR activity. Then, 50 ns simulations of top hit compounds were performed to elucidate the stability of their binding conformations with the LasR-LBD. This study, thus concluded that sulfamerazine showed the highest binding affinity for the LasR-LBD binding pocket exhibiting strong inhibitory binding interactions during molecular dynamics (MD) simulation.

Is home delivery really preferred? a mixed-methods national study in Pakistan.

BACKGROUND: Pakistan has a high maternal mortality ratio and a low rate of skilled birth attendants (SBAs). To address these two important issues, the Pakistan Maternal Newborn and Child Health (MNCH) programme launched the community midwives (CMW) initiative in 2007. CMWs are supposed to conduct deliveries at community level outside health facilities. The purpose of the current study is to document perceptions about CMWs and preferences for birthing place. METHODS: A mixed-methods study was conducted covering four provinces. For the quantitative survey, households were selected through a multistage sampling technique from rural districts. In 1,450 rural households, preferences of respondents about CMW-conducted deliveries were recorded. Qualitative data were obtained through focus group discussions (FGDs) and in-depth interviews (IDIs) with women, community elders, CMWs, and MNCH programme personnel in the same areas where the quantitative study was carried out. In both studies, preferences and the reasons behind particular respondent preferences were recorded. Frequencies of responses were analysed for the quantitative study. Narration and quotes from various types of participants were used to present findings from FGDs and IDIs. RESULTS: In the quantitative study, 42% of respondents expressed a preference for birthing stations, i.e. a place where CMWs conduct deliveries; 22% preferred home deliveries. Birthing stations were favoured because of the availability of space and equipment and the proximity to women's homes. These findings were largely supported by the qualitative component, although a range of views about where a CMW should conduct deliveries were expressed. CONCLUSION: Insights into where CMWs might provide delivery services were obtained through this study. Birthing stations may be an option as a preferred location for delivery care and should be considered as part of Pakistan's national CMW programme.

Expression analysis of ABCA1 in type 2 diabetic Pakistani patients with and without dyslipidemia and correlation with glycemic index and lipid profile.

Diabetes Mellitus type II, earlier considered as an endocrinological disorder is now more regarded as an inflammatory disorder along with lipid aberrations. It demands for regular monitoring, healthy dietary habits and lifestyle modification. This study was focused on gene expression of ATP binding cassette protein 1 (ABCA1) in diabetic dyslipidemia patients in comparison with control groups of only diabetics and healthy individuals. Blood samples and data were collected from recruited 390 patients who were further divided into three groups (130 each). Glycemic index and lipid profile was assessed. Delta Delta Ct method was used that revealed downregulation of the studied gene more in diabetic dyslipidemia patients as compared to only diabetics and healthy controls. The Ct values of ABCA1 were associated with glycemic index and lipid profile using Pearson's correlation. A negative correlation with fasting blood sugar and a positive correlation with HbA1cwas observed in only diabetics group. While in diabetic dyslipidemia and normal healthy controls, a negative correlation was found with both. As far as the lipid profile is concerned a positive correlation was observed among only diabetics with whole lipid profile. In diabetics with dyslipidemia, a negative correlation with all parameters except the TAGs was observed. A positive correlation with all except HDL was observed in healthy controls. The Ct values and fold change were compared among diseased and healthy individuals by applying independent t test. The cycle threshold in only diabetics was p = 0.000018 and in diabetic dyslipdemia individuals was p = 0.00251 while fold change in only diabetics (p = 0.000230) and in diabetics with dyslipidemia (p = 0.001137) was observed to be as statistically significant.

Water Quality Index and Human Health Risk Assessment of Drinking Water in Selected Urban Areas of a Mega City.

The present study was conducted to evaluate the quality of drinking water and assess the potential health hazards due to water contaminants in selected urban areas of Lahore, Pakistan. Water samples were collected from ten sites and analyzed for different physico-chemical parameters including turbidity, color, pH, total dissolved solids (TDS), nitrates, fluoride, residual chlorine, and total hardness. Additionally, heavy metal (arsenic) and microbial parameters (E. coli) were also determined in the water samples. Drinking water quality evaluation indices, including the water quality index (WQI) for physico-chemical and biological parameters and human health risk assessment (HHRA) for heavy metal were estimated using the analytical results of the target parameters. It was found in most of the areas that the levels of arsenic, fluoride, TDS, and residual chlorine were higher than those recommended by the National Environmental Quality Standard (NEQS) and World Health Organization (WHO) guidelines. In addition to the physico-chemical parameters, microbial content (E. coli) was also found in the drinking water samples of the selected areas. Statistical analysis of the results indicated that levels of target parameters in drinking water samples are significantly different between sampling sites. The WQI for all physico-chemical and microbial parameters indicated that drinking water in most of the areas was unfit and unsuitable (WQI > 100) for drinking purposes except for the water of Bhatti Gate and Chota Gaon Shahdara with a WQI of 87 and 91, respectively. Drinking water in these areas had a very poor WQI rating. According to HHRA, drinking water from the selected sites was found to be of high risk to children and adults. The carcinogenic risk of arsenic indicated that all samples were of high risk to both adults and children (4.60 and 4.37 × 10-3, respectively). Regular monitoring of drinking water quality is essential, and proactive measures must be implemented to ensure the treatment and availability of safe drinking water in urban areas.

Glossopharyngeal Neuralgia Associated With Bradycardia and syncope-a Case Report.

We describe a case of 76-year-old woman with glossopharyngeal neuralgia who developed bradycardia and syncope after decreased carbamazepine dosing due to worsening renal function. Telemetry and EKG showed bradycardia and sinus pauses associated with paroxysms of typical glossopharyngeal neuralgia pain. With the addition of gabapentin to carbamazepine, her glossopharyngeal neuralgia pain as well as bradycardia resolved. A pacemaker was placed to prevent bradycardia and syncope. Clinicians should be mindful of the association between glossopharyngeal neuralgia and bradycardia and cardiac syncope so appropriate treatment can be offered in a timely manner to prevent adverse outcomes associated with syncope and cardiac arrest.

Effect of cell density on the biological titer and yield of 146S fraction of foot-and-mouth disease virus O in cell suspension.

Foot-and-mouth disease (FMD) is a highly infectious disease of cattle and other cloven-hoofed animals, causing huge economic losses annually worldwide. This disease is endemic in Pakistan where the serotypes of the foot-and-mouth disease virus (FMDV) are A, O and ASIA-1. At present, trivalent FMDV vaccines are being used to prevent FMD but the current production process is laborious and is unable to fulfill the needs of the meat and dairy industries. To meet the vaccine needs of Pakistan, the conventional method of using adherent cell lines to produce the vaccine could be replaced by suspension cell cultures which produce higher yields in less time and less volume. Therefore, the aim of this study was to investigate and optimize some of the factors that affect viable cell density and subsequent virus yield. The relationship between the yield of the 146S fraction and the TCID50 of the virus preparations obtained was also evaluated as a mean to control and check the quality of the vaccine product. The results provided optimized conditions for vaccine production using cell suspensions and showed that there was a linear relationship between TCID50 and 146S fraction yield. Either TCID50 or the 146S fraction yield, or both could be used as parameters for quality monitoring during vaccine production. Using TCID50 reduced the number of steps involved in virus production while measuring 146S fraction yield was useful for quality control. However, more studies are required to evaluate the relative effectiveness of vaccines produced by virus cultures using either TCID50 or 146S fraction as quality monitoring tools.

Epileptiform activity predicts epileptogenesis in cerebral hemorrhage.

This retrospective case-controlled study was performed to evaluate whether Epileptiform Activity, suspected clinical seizures, and/or 2HELPS2B/S after nontraumatic Intraparenchymal Hemorrhage or Subarachnoid Hemorrhage can predict Epilepsy. Hundred and thirty-two patients were included-29 (Epilepsy), 103 (Control Group). After matching, the average effect for all three risk factors was significant as follows: (1) Epileptiform Activity (p = 0.012, odds ratio 3.14), (2) suspected seizures (p = 0.021, odds ratio 3.78), and (3) 2HELPS2B/S score (p < 0.001, odds ratio 4.94). This study shows that Epileptiform Activity, suspected seizures, and particularly, the 2HELPS2B/S score in the acute phase are risk factors for the development of epilepsy after nontraumatic intraparenchymal and subarachnoid hemorrhage.

Effect of Flavonoid-Coated Gold Nanoparticles on Bacterial Colonization in Mice Organs.

Multidrug resistance (MDR) has been a potentiator for the exploration of antibiotics. Nano drug delivery systems have opened new avenues to overcome this challenge. Although antibacterial nanocarriers are extensively realized, their effect on the bacteria residing inside the tissues and their toxicity is rarely explored. This study investigated the effects of flavonoid coated gold nanoparticles (FAuNPs) on the colonization of Enterococcus faecalis in the mouse liver and kidneys. Flavonoids were extracted from the leaves of Berberis lycium Royle and used to stabilize gold following a green synthesis approach. FAuNPs were characterized by ultraviolet-visible (UV-Vis) spectroscopy, Fourier-transform infrared spectroscopy (FTIR), scanning transmission electron microscopy (STEM), X-ray powder diffraction (XRD), and energy-dispersive X-ray spectroscopy (EDS). FAuNPs showed significantly higher reduction in bacterial counts in in-vitro and in-vivo in mice organs as compared to the free flavonoids owing to their biocompatibility and effectiveness.

Cost analytic model to determine the least costly inpatient erythropoiesis stimulating therapy regimen.

BACKGROUND: Unlike in outpatient settings, the comparative costs of epoetin alpha (EPO) and darbepoetin alpha (DARB) have not been evaluated broadly from the inpatient hospital perspective. OBJECTIVE: To develop a cost analytic model comparing hospital inpatient costs for erythropoiesis stimulating therapies within the nephrology and oncology settings. METHODS: A cost analytic model incorporating erythropoietic drug, pharmacy, and nursing costs was developed from the inpatient hospital perspective to evaluate comparative costs of EPO and DARB. Erythropoietic drug costs were calculated using unit wholesale acquisition cost multiplied by the number of units or micrograms while comparing the following dosing regimens: EPO 3 times weekly, EPO once weekly, and DARB once weekly. Pharmacy costs included dispensing and delivery costs, while nursing costs incorporated administration time costs; all were calculated by estimated fractional hours per activity multiplied by hourly wages. The total frequency of erythropoiesis stimulating therapy administrations was determined based on the average hospital length of stay. The first erythropoiesis stimulating therapy dose was assumed to occur on day 3 of hospitalization. For total inpatient costs, a weighted average was calculated across disease states. One-way sensitivity analyses were conducted by varying length of stay, day of initial erythropoiesis stimulating therapy dose, pharmacy and nursing costs, and once-weekly DARB dose. RESULTS: EPO 3 times weekly was the least costly regimen across all disease states evaluated. Threshold analysis indicated that the cost of once-weekly DARB regimens would have to be reduced by 37% to equal the cost of EPO 3 times weekly for an average length of stay. Sensitivity analyses did not considerably affect the results. CONCLUSIONS: EPO 3 times weekly was found to be the least costly erythropoiesis stimulating therapy regimen for nephrology and oncology inpatients for the average length of stay as well as most other lengths of stay considered. Once-weekly EPO was the least costly erythropoiesis stimulating therapy regimen for several other lengths of stay, while once-weekly DARB was never found to be the least costly regimen.

Chemical sensing of Benzo[a]pyrene using Corchorus depressus fluorescent flavonoids.

Plant phytochemicals, such as flavonoids are in use for the development of optical biosensor. Benzo[a]pyrene (B[a]P), is a pervasive environmental and dietary carcinogen. A fluorescent assay is developed using plant isolated flavonoid for the detection of B[a]P. High content saponins are excluded from the flavonoid-containing methanolic extract of Corchorus depressus by implying reduction of silver ions by saponins resulting in formation of silver nanoparticles. Isolated plant flavonoids are used to develop a spectrofluorometric assay for the detection of B[a]P. Decrease in the flavonoid fluorescence intensity by B[a]P is found to be based on both static and dynamic quenching. Specificity of the assay for B[a]P was tested for other carcinogens belonging to different classes of compounds. Flavonoids-mediated sensing can be implied for the development of new generation of nanoparticle-based biosensors that can be more sensitive and less susceptible to external factors, such as temperature and humidity.

Sequence analysis of hepatitis C virus nonstructural protein 3-4A serine protease and prediction of conserved B and T cell epitopes.

The hepatitis C virus (HCV) is a global health issue. The nonstructural protein 3 (NS3)-4Agene of HCV is responsible for serine protease activity. The aim of the present study was to develop a global consensus sequence of HCV serine protease, analyze conserved residues, and predict highly conserved B- and T-cell binding epitopes in the NS3-4A protein. A total of 160 NS3-4A sequences from the six genotypes of HCV were refracted in the current study. The amino acid sequences were aligned to obtain a global consensus sequence. The location of possible B- and T-cell epitopes were predicted in the HCV NS3-4A consensus sequence by employing bioinformatics tools. Despite the high mutation rate of HCV, the functionally important residues are highly conserved. These include residues that form the catalytic triad (His57, Asp81 and Ser139), the S1 and S6 pocket, zinc-binding site (Cys97, Cys99, Cys145 and His149) and the substrate binding groove. The epitopes B1, B8 and B9 are predicted to be ideal candidates for B-cell-based vaccine and are >95% conserved across six major HCV genotypes. The major histocompatibility complex (MHC) class I epitopes, M4, M5, M7 and M10, and MHC class II epitopes, T5, T7 and T10 are ideal epitopes for vaccine development with high antigenicity scores and high conservancy across major HCV genotypes. The predicted B- and T-cell epitopes are ideal targets for vaccine development, and are capable of producing a strong immune response for all major genotypes of HCV.

PEG capped methotrexate silver nanoparticles for efficient anticancer activity and biocompatibility.

BACKGROUND: Nanocarriers endow tremendous benefits to the drug delivery systems depending upon the specific properties of either component. These benefits include, increase in the drug blood retention time, reduced efflux, additional toxicity and targeted delivery. Methotrexate (MTX) is clinically used for cancer treatment. Higher dosage of MTX results in hepatic and renal toxicity. In this study methotrexate silver nanoparticles (Ag-MTX) coated with polyethylene glycol (PEG) are synthesized and characterized. Their anticancer activity and biocompatibility is also evaluated. RESULTS: Ag-MTX nanoparticles are synthesized by chemical reduction method. They are characterized by Ultraviolet-Visible Spectroscopy and Fourier Transform Infrared Spectroscopy. Average size of PEG coated Ag-MTX nanoparticles (PEG-Ag-MTX nanoparticles) is 12nm. These particles exhibited improved anticancer activity against MCF-7 cell line. Hemolytic activity of these particles was significantly less than MTX. CONCLUSION: PEG-Ag-MTX nanoparticles are potential nanocarrier of methotrexate which may offer MTX based cancer treatment with reduced side effects. In-vivo investigations should be carried out to explore them in detail.

Mixed-linker approach in designing porous zirconium-based metal-organic frameworks with high hydrogen storage capacity.

Three highly porous Zr(iv)-based metal-organic frameworks, UBMOF-8, UBMOF-9, and UBMOF-31, were synthesized by using 2,2'-diamino-4,4'-stilbenedicarboxylic acid, 4,4'-stilbenedicarboxylic acid, and combination of both linkers, respectively. The mixed-linker UBMOF-31 showed excellent hydrogen uptake of 4.9 wt% and high selectivity for adsorption of CO2 over N2 with high thermal stability and moderate water stability with permanent porosity and surface area of 2552 m(2) g(-1).

Intrathoracic gossypiboma.

Gossypiboma refers to retained sponge or swab in any body cavity after surgery. Although it is a rare occurence, it can lead to various complications which include adhesions, abscess formation and subsequent infections. Gossypiboma occurs as a result of not using radio-opaque sponges, poorly performed sponge counts, inadequate wound explorations on suspicion and misread intraoperative radiographs. Therefore, this event can be avoided if strict preventive measures are taken. Moreover, further complications can be avoided following the correct and early diagnosis of gossypiboma. Gossypiboma is an important topic as it carries great medicolegal consequences for the surgeon. We have presented three cases of intrathoracic gossipiboma following previous cardiothoracic surgeries. They presented years after their surgeries, with features varying from patient to patient, ranging from cough and fever to no sypmtoms at all. CT scan only showed a mass lesion in all cases, therefore we proceeded for CT-guided biopsy which was also found to be inconclusive. It was only after entering the thoracic cavity via video-assisted thoracoscopy/thoracotomy that the diagnosis was made and sponges were taken out successfully. All our cases recovered with no further complications.