Outcome of tailored therapy in rheumatic heart disease with persistent atrial fibrillation (RHD-AF).

INTRODUCTION: Rheumatic heart disease with persistent atrial fibrillation (RHD-AF) is associated with increased morbidity. However, there is no standardized approach for the maintenance of sinus rhythm (SR) in them. We aimed to determine the utility of a stepwise approach to achieve SR in RHD-AF. METHODS: Consecutive patients with RHD-AF from July 2021 to August 2023 formed the study cohort. The stepwise approach included pharmacological rhythm control and/or electrical cardioversion (Central illustration). In patients with recurrence, additional options included AF ablation or pace and ablate strategy with conduction system pacing or biventricular pacing. Clinical improvement, NT-proBNP, 6-Minute Walk Test (6MWT), heart failure (HF) hospitalizations, and thromboembolic complications were documented during follow-up. RESULTS: Eighty-three patients with RHD-AF (mean age 56.13 ± 9.51 years, women 72.28%) were included. Utilizing this approach, 43 (51.81%) achieved and maintained SR during the study period of 11.04 ± 7.14 months. These patients had improved functional class, lower NT-proBNP, better distance covered for 6MWT, and reduced HF hospitalizations. The duration of AF was shorter in patients who achieved SR, compared to those who remained in AF (3.15 ± 1.29 vs 6.93 ± 5.23, p = 0.041). Thirty-five percent (29) maintained SR after a single cardioversion over the study period. Only one underwent AF ablation. Of the 24 who underwent pace and ablate strategy, atrial lead was implanted in 22 (hybrid approach), and 50% of these achieved and maintained SR. Among these 24, none had HF hospitalizations, but patients who maintained SR had further improvement in clinical and functional parameters. CONCLUSIONS: RHD-AF patients who could achieve SR with a stepwise approach, had better clinical outcomes and lower HF hospitalizations.

Risk of Distal Embolization From tPA (Tissue-Type Plasminogen Activator) Administration Prior to Endovascular Stroke Treatment.

BACKGROUND AND PURPOSE: In large artery occlusion stroke, both intravenous (IV) tPA (tissue-type plasminogen activator) and endovascular stroke treatment (EST) are standard-of-care. It is unknown how often tPA causes distal embolization, in which a procedurally accessible large artery occlusion is converted to a more distal and potentially inaccessible occlusion. METHODS: We analyzed data from a decentralized stroke telemedicine program in an integrated healthcare delivery system covering 21 hospitals, with 2 high-volume EST centers. We captured all cases sent for EST and examined the relationship between IV tPA administration and the rate of distal embolization, the rate of target recanalization (modified Treatment in Cerebral Infarction scale 2b/3), clinical improvement before EST, and short-term and long-term clinical outcomes. RESULTS: Distal embolization before EST was quite common (63/314 [20.1%]) and occurred more often after IV tPA before EST (57/229 [24.9%]) than among those not receiving IV tPA (6/85 [7.1%]; P<0.001). Distal embolization was associated with an inability to attempt EST: after distal embolization, 26/63 (41.3%) could not have attempted EST because of the new clot location, while in cases without distal embolization, only 8/249 (3.2%) were unable to have attempted EST (P<0.001). Among patients who received IV tPA, 13/242 (5.4%) had sufficient symptom improvement that a catheter angiogram was not performed; 6/342 (2.5%) had improvement to within 2 points of their baseline NIHSS. At catheter angiogram, 2/229 (0.9%) of patients who had received tPA had complete recanalization without distal embolization. Both IV tPA and EST recanalization were associated with improved long-term outcome. CONCLUSIONS: IV tPA administration before EST for large artery occlusion is associated with distal embolization, which in turn may reduce the chance that EST can be attempted and recanalization achieved. At the same time, some IV tPA-treated patients show symptomatic improvement and complete recanalization. Because IV tPA is associated with both distal embolization and improved long-term clinical outcome, there is a need for prospective clinical trials testing the net benefit or harm of IV tPA before EST.

Synthesis of green marine algal-based biochar for remediation of arsenic(V) from contaminated waters in batch and column mode of operation.

The sorption behavior of biochar derived from green seaweed (Ulva reticulata) toward arsenic(V) ions was explored in both batch and continuous modes. The pH edge experiments indicated optimum arsenic(V) sorption observed at pH 4, with maximum sorptional capacity of 7.67 mg/g through isotherm experiments. The kinetic experimental trials indicated that arsenic(V) sorption onto biochar was a fast electrostatic attraction process, with maximum removal occurred within 30 min. The sorption isotherms were modeled using the Toth, Redlich-Peterson, Langmuir and Freundlich isotherm models while the adsorption kinetics was modeled using the pseudo-first- and pseudo-second-order kinetic equations. The three-parameter models (Redlich-Peterson and Toth) better described the isotherm data, whereas pseudo-first-order model represented kinetic data well with low error and high correlation coefficient values. Among the different alkaline and acidic elutants investigated, the solution of 0.01 M NaOH effectively desorbed arsenic(V) from spent biochar. The feasibility of the biochar in continuous remediation of arsenic(V) from contaminated waters was explored in an up-flow fixed column. The biochar exhibited arsenic(V) removal efficiency and sorptional uptake of 59.5% and 8.12 mg/g, respectively. The biochar-loaded column was effectively desorbed using NaOH (0.01 M), with desorption efficiency of 99.5%.

Structural and luminescence properties of Sm3+ -doped bismuth phosphate glass for orange-red photonic applications.

In the present study, the effect of bismuth oxide (Bi2 O3 ) content on the structural and optical properties of 0.5Sm3+ -doped phosphate glass and the effect of concentration on structural and optical properties of Sm3+ -doped bismuth phosphate (BiP) glass were studied. Structural characterization was accomplished using X-ray diffraction (XRD), scanning electron microscopy (SEM) with energy dispersive spectroscopy (EDS), Fourier transform infrared (FTIR) spectroscopy and 31 P nuclear magnetic resonance (NMR) spectroscopy. Optical properties were studied using absorption, photoluminescence and decay measurements. Using optical absorption spectra, Judd-Ofelt parameters were derived to determine the local structure and bonding in the vicinity of Sm3+ ions. The emission spectra of Sm3+ -doped BiP glass showed two intense emission bands, 4 G5/2 →6 H7/2 (orange) and 4 G5/2 →6 H9/2 (red) for which the stimulated emission cross-sections (σe ) and branching ratios (ß) were found to be higher. The quantum efficiencies were also calculated from decay measurements recorded for the 4 G5/2 level of Sm3+ ions. The suitable combination of Bi2 O3 (10 mol%) and Sm3+ (0.5 mol%) ions in these glasses acted as an efficient lasing material and might be suitable for the development of visible orange-red photonic materials.

Concentration-dependent studies of Nd3+ -doped zinc phosphate glasses for NIR photoluminescence at 1.05 µm.

Nd3+ -doped lead-free zinc phosphate glasses with the chemical compositions (60-x) NH4 H2 PO4  + 20ZnO + 10BaF2  + 10NaF + xNd2 O3 (where x = 0.5, 1.0, 1.5, 2.0 and 2.5 mol%) were prepared using a melt quenching technique. Vibrational bands were assigned and clearly elucidated by Raman spectral profiles for all the glass samples. Judd-Ofelt (J-O) intensity parameters (Ωλ : λ = 2, 4, 6) were obtained from the spectral intensities of different absorption bands of Nd3+ ions. Radiative properties such as radiative transition probabilities (AR ), radiative lifetimes (τR ) and branching ratios (ßR ) for different excited states were calculated using J-O parameters. The near infrared (NIR) photoluminescence spectra exhibited three emission bands (4 F3/2 level to 4 I13/2 , 4 I11/2 and 4 I9/2 states) for all the concentrations of Nd3+ ions. Various luminescence properties were studied by varying the Nd3+ concentration for the three spectral profiles. Fluorescence decay curves of the 4 F3/2 level were recorded. The energy transfer mechanism that leads to quenching of the 4 F3/2 state lifetimes was discussed at higher concentration of Nd3+ ions. These glasses are suggested as suitable hosts to produce efficient lasing action in NIR region at 1.05 µm.

Spectral investigations on Eu3+ ,Sm3+ -doped and Sm3+ /Eu3+ co-doped potassium-fluoro-phosphate glass emitting intense orange-red for lighting applications.

Potassium fluoro-phosphate (KFP) glass singly doped with different concentrations of europium (Eu3+ ) or samarium (Sm3+ ) or co-doped (Sm3+ /Eu3+ ) was prepared, and their luminescence spectra were investigated. The phase composition of the product was verified by X-ray diffraction analysis. Optical transition properties of Eu3+ in the studied potassium phosphate glass were evaluated in the framework of the Judd-Ofelt theory. The radiative transition rates (AR ), fluorescence branching ratios (ß), stimulated emission cross-sections (σe ) and lifetimes (τexp ) for certain transitions or levels were evaluated. Red emission of Eu3+ was exhibited mainly by the 5 D0 →7 F2 transition located at 612 nm. Concentration quenching and energy transfer were observed from fluorescence spectra and decay curves, respectively. It was found that the lifetimes of the 5 D0 level increased with increase in concentration and then decreased. By co-doping with Sm3+ , energy transfer from Sm3+ to Eu3+ occurred and contributed to the enhancement in emission intensity. Intense orange-red light emission was obtained upon sensitizing with Sm3+ in KFP glass. This approach shows significant promise for use in reddish-orange lighting applications. The optimized properties of the Sm3+ /Eu3+ co-doped potassium phosphate glass might be promising for optical materials.

Extreme Expression of DNA Repair Protein Apurinic/Apyrimidinic Endonuclease 1 (APE1) in Human Breast Cancer As Measured by Liquid Chromatography and Isotope Dilution Tandem Mass Spectrometry.

Apurinic/apyrimidinic endonuclease 1 (APE1) is a DNA repair protein and plays other important roles. Increased levels of APE1 in cancer have been reported. However, available methods for measuring APE1 levels are indirect and not quantitative. We previously developed an approach using liquid chromatography and tandem mass spectrometry with isotope dilution to accurately measure APE1 levels. Here, we applied this methodology to measure APE1 levels in normal and cancerous human breast tissues. Extreme expression of APE1 in malignant tumors was observed, suggesting that breast cancer cells may require APE1 for survival. Accurate measurement of APE1 may be essential for the development of novel treatment strategies and APE1 inhibitors as anticancer drugs.

The fused anthranilate synthase from Streptomyces venezuelae functions as a monomer.

Recently, we showed that the fused chorismate-utilizing enzyme from the antibiotic-producing soil bacterium Streptomyces venezuelae is an anthranilate synthase (designated SvAS), not a 2-amino-2-deoxyisochorismate (ADIC) synthase, as was predicted based on its amino acid sequence similarity to the phenazine biosynthetic enzyme PhzE (an ADIC synthase). Here, we report the characterization of SvAS using steady-state kinetics, gel filtration chromatography, and laser light scattering. The recombinant His-tagged enzyme has Michaelis constants Km with respect to substrates chorismate and glutamine of 8.2 ± 0.2 µM and 0.84 ± 0.05 mM, respectively, and a catalytic rate constant k cat of 0.57 ± 0.02 s(-1) at 30 °C. Unlike most other anthranilate synthases, SvAS does not utilize ammonia as a substrate. The enzyme is competitively but non-cooperatively inhibited by tryptophan (K i = 11.1 ± 0.1 µM) and is active as a monomer. The finding that SvAS is a monomer jibes with the variety of association modes that have been observed for anthranilate synthases from different microorganisms, and it identifies the enzyme's minimal functional unit as a single TrpE-TrpG pair.

Genomic study in Mexicans identifies a new locus for triglycerides and refines European lipid loci.

BACKGROUND: The Mexican population and others with Amerindian heritage exhibit a substantial predisposition to dyslipidemias and coronary heart disease. Yet, these populations remain underinvestigated by genomic studies, and to date, no genome-wide association (GWA) studies have been reported for lipids in these rapidly expanding populations. METHODS AND FINDINGS: We performed a two-stage GWA study for hypertriglyceridemia and low high-density lipoprotein cholesterol (HDL-C) in Mexicans (n=4361), and identified a novel Mexican-specific genome-wide significant locus for serum triglycerides (TGs) near the Niemann-Pick type C1 protein gene (p=2.43×10(-08)). Furthermore, three European loci for TGs (APOA5, GCKR and LPL), and four loci for HDL-C (ABCA1, CETP, LIPC and LOC55908) reached genome-wide significance in Mexicans. We used cross-ethnic mapping to narrow three European TG GWA loci, APOA5, MLXIPL, and CILP2 that were wide and contained multiple candidate variants in the European scan. At the APOA5 locus, this reduced the most likely susceptibility variants to one, rs964184. Importantly, our functional analysis demonstrated a direct link between rs964184 and postprandial serum apoAV protein levels, supporting rs964184 as the causative variant underlying the European and Mexican GWA signal. Overall, 52 of the 100 reported associations from European lipid GWA meta-analysis generalised to Mexicans. However, in 82 of the 100 European GWA loci, a different variant other than the European lead/best-proxy variant had the strongest regional evidence of association in Mexicans. CONCLUSIONS: This first Mexican GWA study of lipids identified a novel GWA locus for high TG levels; used the interpopulation heterogeneity to significantly restrict three previously known European GWA signals, and surveyed whether the European lipid GWA SNPs extend to the Mexican population.

Multifunctional characteristics of biosynthesized CoFe2O4@Ag nanocomposite by photocatalytic, antibacterial and cytotoxic applications.

Carissa carandas, a traditional medicinal herb with a high concentration of antioxidant phytochemicals, has been used for thousands of years in the Ayurveda, Unani, and homoeopathic schools of medicine. By employing Carissa carandas bark extract as a reducing and capping agent in green biosynthesis, we extend this conventional application to produce CoFe2O4 and CoFe2O4@Ag nanocomposite. A variety of techniques have been used to characterize the synthesised nanocomposite, including UV-Vis, FTIR, XRD, FESEM, EDX, and BET. The CoFe2O4 and CoFe2O4@Ag nanocomposite demonstrated promising antibacterial action against human bacterial pathogens like B. subtilis and S. aureus as gram positive and P. aeruginosa and E. coli as gram negative with inhibition zones of 24.3 ± 0.57, 17.4 ± 0.75 and 20.5 ± 0.5, 19.8 ± 1.6 mm respectively, and the obtained results were superior to the nanocomposite without silver. Moreover, in-vitro cytotoxicity effects of biosynthesized CoFe2O4 and CoFe2O4@Ag were performed on the human breast cancer cell MCF-7. It was found that the MCF-7 cells' 50% inhibitory concentration (IC50) was 60 µg/mL. Additionally, biosynthesized CoFe2O4 and CoFe2O4@Ag nanocomposite was used to demonstrate the photocatalytic eradication of Rhodamine Blue (RhB). Due to the addition of Ag, which increases surface area, conductivity, and increased charge carrier separation, the CoFe2O4@Ag nanocomposite exhibits a high percentage of photocatalytic degradation of ⁓ 98% within 35 min under UV light irradiation. The photocatalytic performance of as-synthesised nanocomposite was evaluated using dye degradation-adsorption in both natural light and dark condition. Under dark conditions, it was found that 2 mg mL-1 CoFe2O4@Ag in RhB aqueous solution (5 ppm) causes dye adsorption in 30 min with an effectiveness of 72%. Consequently, it is anticipated that the CoFe2O4@Ag nanocomposite will be a promising photocatalyst and possibly a noble material for environmental remediation applications.

Identification and quantification of human DNA repair protein NEIL1 by liquid chromatography/isotope-dilution tandem mass spectrometry.

Accumulated evidence points to DNA repair capacity as an important factor in cancer and other diseases. DNA repair proteins are promising drug targets and are emerging as prognostic and therapeutic biomarkers. Thus, the knowledge of the overexpression or underexpression levels of DNA repair proteins in tissues will be of fundamental importance. In this work, mass spectrometric assays were developed for the measurement in tissues of the human DNA repair protein NEIL1 (hNEIL1), which is involved in base excision and nucleotide excision repair pathways of oxidatively induced DNA damage. Liquid chromatography/isotope-dilution tandem mass spectrometry (LC-MS/MS), in combination with a purified and fully characterized recombinant (15)N-labeled analogue of hNEIL1 ((15)N-hNEIL1) as an internal standard, was utilized to develop an accurate method for the quantification of hNEIL1. Both hNEIL1 and (15)N-hNEIL1 were hydrolyzed with trypsin, and 18 tryptic peptides from each protein were identified by LC-MS/MS on the basis of their full-scan mass spectra. These peptides matched the theoretical peptides expected from trypsin hydrolysis of hNEIL1 and provided a statistically significant protein score that would unequivocally identify hNEIL1. The product ion spectra of the tryptic peptides from both proteins were recorded, and the characteristic product ions were defined. Selected-reaction monitoring was used to analyze mixtures of hNEIL1 and (15)N-hNEIL1 on the basis of product ions. Additional confirmation of positive identification was demonstrated via separation of the proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and in-gel tryptic digestion followed by LC-MS/MS analysis. These results suggest that the developed assays would be highly suitable for the in vivo positive identification and accurate quantification of hNEIL1 in tissues.

Analysis of chemical engineering curriculum to improve process safety competency.

Continuous process safety (PS) development is the key to maintaining a good PS system, and its competency plays a substantial role. However, PS incompetency can still be demonstrated in several process-related accidents, particularly major catastrophic incidents. To mitigate this gap, universities' PS education is analysed. Because PS is an important element of chemical engineering (CE), this study seeks to identify the most prevalent PS subjects taught in the top 300 Quacquarelli Symonds ranking (2019) universities. Findings indicate that PS education remains insufficiently addressed in undergraduate CE curricula over the years. Twelve common topics, i.e., human factors; management of hazards, incidents, and risk; design; fire and explosion; legislation and standards; sustainability; process control; economics; toxicology; and software are identified. Notably, sustainability is acknowledged to be a new common PS topic, depicting its demand for industrial evolution. Ultimately, strengthening the collaboration between universities and industries is required to develop graduates' PS competency.Abbreviations: ALARP: as low as reasonably practicable; CAD: computer-aided design; CE: chemical engineering; ETA: event tree analysis; FTA: fault tree analysis; FMEA: failure mode and effect analysis; HAZAN: hazard analysis; HAZID: hazard identification; HAZOP: hazard and operability; HSE: health, safety and environment; HYSYS: Hyprotech Systems; LCA: life cycle analysis; LOPA: layer of protection analysis; MS: Microsoft; ORP: occupational risk prevention; PC: personal computer; PHA: process hazard analysis; PS: process safety; PSM: process safety management; QS: Quacquarelli Symonds; SMS: safety management system.

An ultra-sensitive laccase/polyaziridine-bismuth selenide nanoplates modified GCE for detection of atenolol in pharmaceuticals and urine samples.

The analysis of ß-blockers in pharmaceutical, biological and environmental samples has gained much interest due to their wide applications. The aim of this study was to develop an enzyme-based biosensor using hexagonal-shaped low-dimensional Bi2Se3 NPs decorated with laccase through polyaziridine (PAZ) modified glassy carbon electrode (Lac/PAZ-Bi2Se3 NPs/GCE). Surface properties were examined using SEM, TEM, EDX, XRD, XPS, FTIR, UV-Visible, and zeta potential. Electrochemical studies were performed with cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The enzymatic biosensor exhibited excellent catalytic activity towards the oxidation of ATN at +1.05 V (vs Ag/AgCl). Under the optimum experimental conditions, Ip (µA) was linearly related to the concentrations of ATN in the range of 3 to 130 µM (R2 = 0.9972) with an LOD of 0.15 µM and 0.21 µM with and without Lac enzyme. Additionally, the validation of the biosensor was tested to determine ATN on within a day and between-day basis. The biosensor was applied successfully to detect ATN in real samples. The obtained recoveries range from 98.5 % to 99.2 % with an RSD (n = 5) of 0.95 (±0.02). The findings of this study have potential biomedical applications in drug detection employing a promising nano electrode sensor of Lac/PAZ-Bi2Se3 NPs/GCE.

The potential of polyethylene microplastics to transport copper in aquatic systems: Adsorption and desorption studies.

Heavy metals and microplastics are two types of general contaminants that can cause significant damage to water systems and organisms. However, the interaction of heavy metal ions with microplastic in aquatic systems received less attention compared with that of organic contaminants. This study aims to explore the interaction of copper (II) ions with microplastic (polyethylene) in aquatic systems. The adsorption experiments were performed by varying several operational parameters including equilibrium pH, initial Cu (II) concentrations, and contact times. The microplastic was characterized using X-ray diffraction, scanning electron microscopy, and Fourier transform infrared. The results confirmed the porous nature of the microplastic surface and the existence of various binding sites. The maximum Cu (II) uptake by microplastic was recorded as 1.23 mg/g at pH 5, according to the Langmuir adsorption isotherm. The experimental isotherm data exhibited a good fit to the Toth model, followed by the Langmuir and Freundlich equations, according to the correlation coefficient and %error values. The pseudo-first kinetics equation showed a better fit to copper (II) kinetics data compared with the pseudo-second kinetics equation. Elution of copper (II) ions from copper (II)-loaded microplastic was attempted using several elutants, and the results indicated that 0.01 M HNO3 performed well with elution efficiency over 99.5%. Thus, the elution experiments furnished proof that Cu-loaded microplastic may leach Cu (II) ions under rich acidic conditions, thereby aiding the transport of Cu (II) ions into the digestive tracts of aquatic organisms. PRACTITIONER POINTS: Polyethylene microplastics showed potential to sorb copper ions. The mechanism was electrostatic interaction between microplastics and metal ions. Maximum copper adsorption by microplastic was recorded as 1.23 mg/g. Once desorbed, Cu(II) transferred into the digestive tracts of aquatic organisms.

Identification and quantification of DNA repair proteins by liquid chromatography/isotope-dilution tandem mass spectrometry using their fully 15N-labeled analogues as internal standards.

Oxidatively induced DNA damage is implicated in disease, unless it is repaired by DNA repair. Defects in DNA repair capacity may be a risk factor for various disease processes. Thus, DNA repair proteins may be used as early detection and therapeutic biomarkers in cancer and other diseases. For this purpose, the measurement of the expression level of these proteins in vivo will be necessary. We applied liquid chromatography/isotope-dilution tandem mass spectrometry (LC-MS/MS) for the identification and quantification of DNA repair proteins human 8-hydroxyguanine-DNA glycosylase (hOGG1) and Escherichia coli formamidopyrimidine DNA glycosylase (Fpg), which are involved in base-excision repair of oxidatively induced DNA damage. We overproduced and purified (15)N-labeled analogues of these proteins to be used as suitable internal standards to ensure the accuracy of quantification. Unlabeled and (15)N-labeled proteins were digested with trypsin and analyzed by LC-MS/MS. Numerous tryptic peptides of both proteins were identified on the basis of their full-scan mass spectra. These peptides matched the theoretical peptide fragments expected from trypsin digestion and provided statistically significant protein scores that would unequivocally identify these proteins. We also recorded the product ion spectra of the tryptic peptides and defined the characteristic product ions. Mixtures of the analyte proteins and their (15)N-labeled analogues were analyzed by selected-reaction monitoring on the basis of product ions. The results obtained suggest that the methodology developed would be highly suitable for the positive identification and accurate quantification of DNA repair proteins in vivo as potential biomarkers for cancer and other diseases.

Stable isotope-labeling of DNA repair proteins, and their purification and characterization.

Reduced DNA repair capacity is associated with increased risk for a variety of disease processes including carcinogenesis. Thus, DNA repair proteins have the potential to be used as important predictive, prognostic and therapeutic biomarkers in cancer and other diseases. The measurement of the expression level of these enzymes may be an excellent tool for this purpose. Mass spectrometry is becoming the technique of choice for the identification and quantification of proteins. However, suitable internal standards must be used to ensure the precision and accuracy of measurements. An ideal internal standard in this case would be a stable isotope-labeled analog of the analyte protein. In the present work, we over-expressed, purified and characterized two stable isotope-labeled DNA glycosylases, i.e., (15)N-labeled Escherichia coli formamidopyrimidine DNA glycosylase (Fpg) and (15)N-labeled human 8-oxoguanine-DNA glycosylase (hOGG1). DNA glycosylases are involved in the first step of the base excision repair of oxidatively induced DNA damage by removing modified DNA bases. The measurement by MALDI-ToF mass spectrometry of the molecular mass and isotopic purity proved the identity of the (15)N-labeled proteins and showed that the (15)N-labeling of both proteins was more than 99.7%. We also measured the DNA glycosylase activities using gas chromatography/mass spectrometry with isotope-dilution. The enzymic activities of both (15)N-labeled Fpg and (15)N-labeled hOGG1 were essentially identical to those of their respective unlabeled counterparts, ascertaining that the labeling did not perturb their catalytic sites. The procedures described in this work may be used for obtaining stable isotope-labeled analogs of other DNA repair proteins for mass spectrometric measurements of these proteins as disease biomarkers.

Fusing an insoluble protein to GroEL apical domain enhances soluble expression in Escherichia coli.

A protocol for increasing soluble protein expression by fusing the chaperone GroEL apical domain with a gene of interest is described herein. GroEL apical domain, the minichaperone that functions independently of GroES and ATP in protein folding, is cloned downstream of the lambda CII ribosome binding site in the parent pRE vector. The pRE vector has tightly controlled transcription suitable for expressing toxic proteins. The GroEL minichaperone is fused to a glycine-serine rich linker followed by the enterokinase protease recognition sequence. A number of genes that are recalcitrant to protein production in the parent pRE vector 5were cloned into the pRE:GroEL fusion vector and successfully expressed as fusion proteins in Escherichia coli.

Predictors of major neurological improvement after intravenous thrombolysis in acute ischemic stroke: a hospital-based study from south India.

BACKGROUND: Despite the increasing use of recombinant tissue plasminogen activator (rt-PA) in acute ischemic stroke, uncertainty persists about the short- and long-term outcome of the thrombolysed patients. OBJECTIVE: To identify predictors of major neurological improvement at 24 h after intravenous rt-PA administration in patients of acute ischemic stroke and their relationship with outcome at 12 months. MATERIALS AND METHODS: We analyzed the data of the patients with acute ischemic stroke treated as per the National Institute of Neurological Disorders and Stroke (NINDS) criteria with intravenous rt-PA between January 2000 and June 2009 at a tertiary care center in south India. Major neurological improvement was defined by an 8-point improvement in National Institute of Health Stroke Scale (NIHSS) score or an NIHSS score of 0 or 1 at 24 h. Good outcome was defined as a 12-month modified Rankin Scale (mRS) of 0 to 1. RESULTS: Of the 72 patients with acute ischemic stroke treated with intravenous rt-PA, 23 (32%) patients had major neurological improvement at 24 h. Age <60 years (OR 1.9, 95% CI 1.7 to 3.2), admission glucose levels <8 mmol/L (OR 3.87, 95% CI 1.9 to 9.2) and mild to moderate baseline stroke severity (NIHSS median score 10+ 6) were associated with major neurological improvement after adjusting for co variables. Major neurological improvement at 24 h was an independent predictor of good outcome (mRS=1) at 12 months (OR 13.9, 95% CI 6.84 to 40.2). CONCLUSIONS: Age <60 years, glucose levels <8 mmol/L and mild to moderate stroke severity (NIHSS median score 10+/-6) was associated with major neurological improvement after intravenous rt-PA. Major neurological improvement at 24 h after the administration of intravenous thrombolysis independently predicted good outcome at 12 months.

Resection of Olfactory Groove Meningiomas Through Unilateral vs. Bilateral Approaches: A Systematic Review and Meta-Analysis.

Introduction: Consensus is limited regarding optimal transcranial approaches (TCAs) for the surgical resection of olfactory groove meningiomas (OGMs). This systematic review and meta-analysis aims to examine operative and peri-operative outcomes of unilateral compared to bilateral TCAs for OGMs. Methods: Electronic databases were searched from inception until December 2019 for studies delineating TCAs for OGM patients. Patient demographics, pre-operative symptoms, surgical outcomes, and complications were evaluated and analyzed with a meta-analysis of proportions. Results: A total of 27 observational case series comparing 554 unilateral vs. 451 bilateral TCA patients were eligible for review. The weighted pooled incidence of gross total resection is 94.6% (95% CI, 90.7-97.5%; I 2 = 59.0%; p = 0.001) for unilateral and 90.9% (95% CI, 85.6-95.4%; I 2 = 58.1%; p = 0.003) for bilateral cohorts. Similarly, the incidence of OGM recurrence is 2.6% (95% CI, 0.4-6.0%; I 2 = 53.1%; p = 0.012) and 4.7% (95% CI, 1.4-9.2%; I 2 = 55.3%; p = 0.006), respectively. Differences in oncologic outcomes were not found to be statistically significant (p = 0.21 and 0.35, respectively). Statistically significant differences in complication rates in bilateral vs. unilateral TCA cohorts include meningitis (1.0 vs. 0.0%; p = 0.022) and mortality (3.2 vs. 0.2%; p = 0.007). Conclusions: While both cohorts have similar oncologic outcomes, bilateral TCA patients exhibit higher post-operative complication rates. This may be explained by underlying tumor characteristics necessitating more radical resection but may also indicate increased morbidity with bilateral approaches. However, evidence from more controlled, comparative studies is warranted to further support these findings.

A comparative biochemical and structural analysis of the intracellular chorismate mutase (Rv0948c) from Mycobacterium tuberculosis H(37)R(v) and the secreted chorismate mutase (y2828) from Yersinia pestis.

The Rv0948c gene from Mycobacterium tuberculosis H(37)R(v) encodes a 90 amino acid protein as the natural gene product with chorismate mutase (CM) activity. The protein, 90-MtCM, exhibits Michaelis-Menten kinetics with a k(cat) of 5.5+/-0.2s(-1) and a K(m) of 1500+/-100microm at 37 degrees C and pH7.5. The 2.0A X-ray structure shows that 90-MtCM is an all alpha-helical homodimer (Protein Data Bank ID: 2QBV) with the topology of Escherichia coli CM (EcCM), and that both protomers contribute to each catalytic site. Superimposition onto the structure of EcCM and the sequence alignment shows that the C-terminus helix3 is shortened. The absence of two residues in the active site of 90-MtCM corresponding to Ser84 and Gln88 of EcCM appears to be one reason for the low k(cat). Hence, 90-MtCM belongs to a subfamily of alpha-helical AroQ CMs termed AroQ(delta.) The CM gene (y2828) from Yersinia pestis encodes a 186 amino acid protein with an N-terminal signal peptide that directs the protein to the periplasm. The mature protein, *YpCM, exhibits Michaelis-Menten kinetics with a k(cat) of 70+/-5s(-1) and K(m) of 500+/-50microm at 37 degrees C and pH7.5. The 2.1A X-ray structure shows that *YpCM is an all alpha-helical protein, and functions as a homodimer, and that each protomer has an independent catalytic unit (Protein Data Bank ID: 2GBB). *YpCM belongs to the AroQ(gamma) class of CMs, and is similar to the secreted CM (Rv1885c, *MtCM) from M.tuberculosis.