Designing a TiO2-MoO3-BMIMBr nanocomposite by a solvohydrothermal method using an ionic liquid aqueous mixture: an ultra high sensitive acetaminophen sensor.

This study shows a simplistic, efficient procedure to synthesize TiO2-MoO3-BMIMBr nanocomposites. Powder X-ray diffraction, scanning electron microscopy, energy-dispersive X-ray spectroscopy, and X-ray photoelectron spectroscopy have all been used to completely analyse the materials. The detection of acetaminophen (AC) has been examined at a modified glassy carbon electrode with TiO2-MoO3-BMIMBr nanocomposites. Moreover, the electrochemical behavior of the nanocomposite modified electrode has been studied by cyclic voltammetry (CV), differential pulse voltammetry (DPV), chronoamperometry and electrochemical impedance spectroscopy (EIS). The linear response of AC was observed in the range 8.26-124.03 nM. The sensitivity and detection limits (S/N = 3) were found to be 1.16 µA L mol-1 cm-2 and 11.54 nM by CV and 24 µA L mol-1 cm-2 and 8.16 nM by DPV respectively.

SARS-CoV-2 variants of concern surveillance including Omicron using RT-PCR-based genotyping offers comparable performance to whole genome sequencing.

Known SARS-CoV-2 variants of concern (VOCs) can be detected and differentiated using an RT-PCR-based genotyping approach, which offers quicker time to result, lower cost, higher flexibility, and use of the same laboratory instrumentation for detection of SARS-CoV-2 when compared with whole genome sequencing (WGS). In the current study, we demonstrate how we applied a genotyping approach for identification of all VOCs and that such technique can offer comparable performance to WGS for identification of known SARS-CoV-2 VOCs, including more recent strains, Omicron BA.1 and BA.2.

Nanoparticle surface energy transfer (NSET) in ferroelectric liquid crystal-metallic-silver nanoparticle composites: Effect of dopant concentration on NSET parameters.

In the recent past, the resonance energy transfer studies using metallic nanoparticles has become a matter of quintessence in modern technology, which considerably extends its applications in probing specific biological and chemical processes. In the present study, metallic-silver nanoparticles of 2-4 nm (diameter) capped with hexanethiol ligand are developed and dispersed in ferroelectric liquid crystal (FLC). The morphology of nanoparticles was characterized using HR-TEM and SEM techniques. Furthermore, a systematic study of energy transfer between the host FLC material (as donors) and metallic-silver nanoparticles (as acceptors) has been explored employing steady state and time resolved fluorescence spectroscopic techniques. The nanoparticle based surface energy transfer (NSET) parameters viz., transfer efficiency, transfer rate, and proximity distance between donor and acceptor, have been determined for NSET couples (FLC material-metallic-silver nanoparticle) composites. It is observed that various NSET parameters and quenching efficiency follow a linear dependence on the concentration of metallic-silver nanoparticles in host FLC material. The nonradiative energy transfer and superquenching effect were analyzed with the help of Stern-Volmer plots. The impact of present study about superquenching effect of the silver nanoparticles can be used for sensing applications that require high degree sensitivity.

Psychiatrist facing litigation.

Malpractice is civil wrong actionable by law. It is true, we have statistically an outside chance of getting sued. Among physicians of all specialties, we have, between 2.6% in North America and 1.3% in Europe, chance of facing malpractice suit. But, that is no comfort for one being sued. Malpractice generally involves, among others, wrongdoing or professional misconduct. It may or may not lead to legal action. I describe two cases of malpractice and its impact on the morale of my mental health team. In one case, the family were seeking compensation by claiming "medical negligence" in a trivial mishap. They had involved the local press to "publicize" their claim. In another case, the grievance arose due to anti-electroconvulsive therapy views of the patient's sister. The families decided not to sue us and accepted decisions of local inquiry. Not much is written on this subject in our country. The cases are almost 20 years old but are still relevant. The author describes the harrowing experience of the team members while the investigation was on. Fortunately, the inquiry panel concluded the proceedings in just 4 weeks. In addition, the author advises how one can prepare the defense. The readers would find informative the case vignettes and strategy to deal with the crisis.

Crystal structure of 4,5,6,7,8,8-hexa-chloro-2-(3,4-di-meth-oxy-pheneth-yl)-3a,4,7,7a-tetra-hydro-1H-4,7-methano-iso-indole-1,3(2H)-dione [+solvent].

In the title compound, C19H15Cl6NO4 [+solvent], the six-membered ring of the norbornene moiety adopts a boat conformation and the two five-membered rings have envelope conformations. The pyrrolidine ring makes a dihedral angle of 14.83 (12)° with the 3,4-di-meth-oxy-phenyl ring, which are attached to each other by an extended N-CH2-CH2-Car bridge. In the crystal, the structure features C-H⋯O inter-molecular hydrogen bonds, an offset π-π inter-action [inter-centroid distance = 3.564 (1) Å] and a C-Cl⋯π inter-action. The contribution of some disordered solvent to the scattering was removed using the SQUEEZE routine [Spek (2015 ▸). Acta Cryst. C71, 9-18] of PLATON. The solvent contribution was not included in the reported mol-ecular weight and density.

Structural insights and binding of a natural ligand, succinic acid with serine and cysteine proteases.

In the age of growing infectious diseases, there is a great demand for new inhibitors which can exhibit minimum side effects. Owing to the importance of proteases in life cycle and invasion, they have been projected as attractive targets for structure based drug designing against microbes including viruses. Here we report the inhibitory activity of a well known natural compound succinic acid against both serine and cysteine proteases. The ligand is found co-crystallized with Bovine pancreatic trypsin in one of our crystallization trials and the diffraction data up to1.9 Å reveal its interactions with the catalytic triad residues Histidine 57 and Serine 195. Binding of the ligand with these proteases have been validated using caseinolysis inhibition. With trypsin, ITC analysis showed tight binding of the ligand, resulting in change in Gibb's free energy (ΔG) by -20.31 kJ/mol. To understand the existence of succinic acid at the active site, molecular docking was performed and it revealed binding of it with trypsin and papain at corresponding active sites. This dual inhibitory activity of natural ligand, succinic acid can be accounted for the recent reports on anti-viral property of plant extracts where dicarboxilic fatty acids are normally abundant.

Using Whole Genome Analysis to Examine Recombination across Diverse Sequence Types of Staphylococcus aureus.

Staphylococcus aureus is an important clinical pathogen worldwide and understanding this organism's phylogeny and, in particular, the role of recombination, is important both to understand the overall spread of virulent lineages and to characterize outbreaks. To further elucidate the phylogeny of S. aureus, 35 diverse strains were sequenced using whole genome sequencing. In addition, 29 publicly available whole genome sequences were included to create a single nucleotide polymorphism (SNP)-based phylogenetic tree encompassing 11 distinct lineages. All strains of a particular sequence type fell into the same clade with clear groupings of the major clonal complexes of CC8, CC5, CC30, CC45 and CC1. Using a novel analysis method, we plotted the homoplasy density and SNP density across the whole genome and found evidence of recombination throughout the entire chromosome, but when we examined individual clonal lineages we found very little recombination. However, when we analyzed three branches of multiple lineages, we saw intermediate and differing levels of recombination between them. These data demonstrate that in S. aureus, recombination occurs across major lineages that subsequently expand in a clonal manner. Estimated mutation rates for the CC8 and CC5 lineages were different from each other. While the CC8 lineage rate was similar to previous studies, the CC5 lineage was 100-fold greater. Fifty known virulence genes were screened in all genomes in silico to determine their distribution across major clades. Thirty-three genes were present variably across clades, most of which were not constrained by ancestry, indicating horizontal gene transfer or gene loss.

Comparative Evaluation of Specific Phytochemical Indicatives in Cirivilvadi Kasaya Prepared Freshly and at Commercial Scale.

Kasaya or decoction is an Ayurvedic dosage form, prescribed based on the stage of the disease according to the principles of Ayurveda. This dosage form is traditionally prepared fresh and consumed on the same day but for the sake of convenience; the process of preparation has been modified so that it can be stored with longer shelf life, easy availability and produced in large quantities. There is a need to understand the implications of this modification in terms of chemical changes. This work attempted to check the phytochemical profile of both freshly prepared decoction and commercially available decoction with reference to some analytical parameters like pH, total soluble solids, phenols, alkaloids, potassium and to assess the changes in the thin layer chromatography profiling of the decoction. The results showed that phenols and potassium are found to be two fold higher in freshly prepared decoction, compared to commercially available decoction diluted to dosage in practice (1:4 ratio). However, the total alkaloid content was found to be approximately ten fold higher in commercially available decoction. It was observed that the thin layer chromatography profile of decoctions was extracted into petroleum ether and chloroform was similar and consistent with different batches though the bands in commercially available decoction were slightly more intense compared to freshly prepared decoction. The total soluble solids in commercially available decoction were four times higher than freshly prepared decoction. The study reveals that there are differences in the phytochemical profiles of the freshly prepared decoction and commercially available decoction of the same formulation. However, the significance of these differences can be determined only by further clinical studies. On the other hand, the study lends support to the practice of diluting the commercially available decoction to make it equivalent to freshly prepared decoction.

Stability of oryzanol fortified biscuits on storage.

Biscuit can be used as a functional food to deliver nutraceuticals to consumers. One such natural nutraceutical oryzanol is present in rice bran oil. Oryzanol possesses a variety of health benefits which include reduction of cholesterol in blood, improvement of capillary action of blood vessels, anti-aging effect and others. Biscuit is a well known cereal based processed food and the fortification of oryzanol into the biscuits will go a long way to provide antioxidant rich, highly stable and acceptable functional food to the consumers. Biscuits were prepared with commercially available fat (CF) and oryzanol fortified fat (OFF). The control biscuits (CB) and oryzanol fortified biscuits (OFB) were packed in 200 gauge polypropylene pouches, stored at 27 °C with different relative humidity (RH 11 %, 22 %, 32 %, 44 % and 56 %) and analysed for its stability during storage of 120 days. Critical moisture content of OFB (4.8 %) was slightly less than that of CB (5.3 %). The fat content of the CB (12.2 %) and OFB (12.5 %) did not change during storage while free fatty acid content (0.36 % and 0.60 %) and peroxide value (0.08 and 0.17 meq. O2/100 g biscuit) respectively for CB and OFFB was showed small but significant changes during storage. Oryzanol content (292 mg) and radical scavenging activity (81.1 %) of OFB did not change during storage. The biscuits had a shelf life of minimum 3 months at 27 °C. Oryzanol in OFB showed good stability during baking and storage of biscuits.

Posological Considerations of Ocimum sanctum (Tulasi) as per Ayurvedic Science and Pharmaceutical Sciences.

A study in 2010 reported that administration of 2 g of O. sanctum leaves for 30 days to laboratory male albino rabbits showed adverse effect on sperm count and male hormones. The dose and duration at which this testing was reported was commented upon as being high. It is learnt that basis this publication a few European regulators have imposed restrictions on usage of O. sanctum. Recognizing the need for evaluation, a review has been made of the posological considerations related to decision on dose of a drug in pharmaceuticals (drug development stages) and in Ayurvedic science as part of history of use and current usage. Specifically, we report the dose range as per documented tradition, marketed products containing O. sanctum as an ingredient and current clinical practice. Greater consultation is suggested before deciding the studies on Ayurvedic herbs. Regulatory action of banning use of O. sanctum needs a review and may need to be replaced with an advisory.

Genomic characterization of Salmonella Cerro ST367, an emerging Salmonella subtype in cattle in the United States.

BACKGROUND: Within the last decade, Salmonella enterica subsp. enterica serovar Cerro (S. Cerro) has become one of the most common serovars isolated from cattle and dairy farm environments in the northeastern US. The fact that this serovar is commonly isolated from subclinically infected cattle and is rarely associated with human disease, despite its frequent isolation from cattle, has led to the hypothesis that this emerging serovar may be characterized by reduced virulence. We applied comparative and population genomic approaches to (i) characterize the evolution of this recently emerged serovar and to (ii) gain a better understanding of genomic features that could explain some of the unique epidemiological features associated with this serovar. RESULTS: In addition to generating a de novo draft genome for one Salmonella Cerro strain, we also generated whole genome sequence data for 26 additional S. Cerro isolates, including 16 from cattle operations in New York (NY) state, 2 from human clinical cases from NY in 2008, and 8 from diverse animal sources (7 from Washington state and 1 from Florida). All isolates sequenced in this study represent sequence type ST367. Population genomic analysis showed that isolates from the NY cattle operations form a well-supported clade within S. Cerro ST367 (designated here "NY bovine clade"), distinct from isolates from Washington state, Florida and the human clinical cases. A molecular clock analysis indicates that the most recent common ancestor of the NY bovine clade dates back to 1998, supporting the recent emergence of this clone.Comparative genomic analyses revealed several relevant genomic features of S. Cerro ST367, that may be responsible for reduced virulence of S. Cerro, including an insertion creating a premature stop codon in sopA. In addition, patterns of gene deletion in S. Cerro ST367 further support adaptation of this clone to a unique ecological or host related niche. CONCLUSIONS: Our results indicate that the increase in prevalence of S. Cerro ST367 is caused by a highly clonal subpopulation and that S. Cerro ST367 is characterized by unique genomic deletions that may indicate adaptation to specific ecological niches and possibly reduced virulence in some hosts.

Targeting c-Met in melanoma: mechanism of resistance and efficacy of novel combinatorial inhibitor therapy.

Numerous tyrosine kinase inhibitors (TKIs) targeting c-Met are currently in clinical trials for several cancers. Their efficacy is limited due to the development of resistance. The present study aims to elucidate this mechanism of c-Met TKI resistance by investigating key mTOR and Wnt signaling proteins in melanoma cell lines resistant to SU11274, a c-Met TKI. Xenografts from RU melanoma cells treated with c-Met TKIs SU11274 and JNJ38877605 showed a 7- and 6-fold reduction in tumor size, respectively. Resistant cells displayed upregulation of phosphorylated c-Met, mTOR, p70S6Kinase, 4E-BP1, ERK, LRP6, and active ß-catenin. In addition, GATA-6, a Wnt signaling regulator, was upregulated, and Axin, a negative regulator of the Wnt pathway, was downregulated in resistant cells. Modulation of these mTOR and Wnt pathway proteins was also prevented by combination treatment with SU11274, everolimus, an mTOR inhibitor, and XAV939, a Wnt inhibitor. Treatment with everolimus, resulted in 56% growth inhibition, and a triple combination of SU11274, everolimus and XAV939, resulted in 95% growth inhibition in RU cells. The V600E BRAF mutation was found to be positive only in MU cells. Combination treatment with a c-Met TKI and a BRAF inhibitor displayed a synergistic effect in reducing MU cell viability. These studies indicate activation of mTOR and Wnt signaling pathways in c-Met TKI resistant melanoma cells and suggest that concurrent targeting of c-Met, mTOR, and Wnt pathways and BRAF may improve efficacy over traditional TKI monotherapy in melanoma patients.

Progress toward an efficient panel of SNPs for ancestry inference.

Many panels of ancestry informative single nucleotide polymorphisms have been proposed in recent years for various purposes including detecting stratification in biomedical studies and determining an individual's ancestry in a forensic context. All of the panels have limitations in their generality and efficiency for routine forensic work. Some panels have used only a few populations to validate them. Some panels are based on very large numbers of SNPs thereby limiting the ability of others to test different populations. We have been working toward an efficient and globally useful panel of ancestry informative markers that is comprised of a small number of highly informative SNPs. We have developed a panel of 55 SNPs analyzed on 73 populations from around the world. We present the details of the panel and discuss its strengths and limitations.

Comparative analysis of genome sequences covering the seven cronobacter species.

BACKGROUND: Species of Cronobacter are widespread in the environment and are occasional food-borne pathogens associated with serious neonatal diseases, including bacteraemia, meningitis, and necrotising enterocolitis. The genus is composed of seven species: C. sakazakii, C. malonaticus, C. turicensis, C. dublinensis, C. muytjensii, C. universalis, and C. condimenti. Clinical cases are associated with three species, C. malonaticus, C. turicensis and, in particular, with C. sakazakii multilocus sequence type 4. Thus, it is plausible that virulence determinants have evolved in certain lineages. METHODOLOGY/PRINCIPAL FINDINGS: We generated high quality sequence drafts for eleven Cronobacter genomes representing the seven Cronobacter species, including an ST4 strain of C. sakazakii. Comparative analysis of these genomes together with the two publicly available genomes revealed Cronobacter has over 6,000 genes in one or more strains and over 2,000 genes shared by all Cronobacter. Considerable variation in the presence of traits such as type six secretion systems, metal resistance (tellurite, copper and silver), and adhesins were found. C. sakazakii is unique in the Cronobacter genus in encoding genes enabling the utilization of exogenous sialic acid which may have clinical significance. The C. sakazakii ST4 strain 701 contained additional genes as compared to other C. sakazakii but none of them were known specific virulence-related genes. CONCLUSIONS/SIGNIFICANCE: Genome comparison revealed that pair-wise DNA sequence identity varies between 89 and 97% in the seven Cronobacter species, and also suggested various degrees of divergence. Sets of universal core genes and accessory genes unique to each strain were identified. These gene sequences can be used for designing genus/species specific detection assays. Genes encoding adhesins, T6SS, and metal resistance genes as well as prophages are found in only subsets of genomes and have contributed considerably to the variation of genomic content. Differences in gene content likely contribute to differences in the clinical and environmental distribution of species and sequence types.

Validation of the Applied Biosystems MicroSEQ real-time PCR system for detection of E. coli O157:H7 in food.

Modern molecular methods offer the advantages of simplicity and short time-to-results compared to traditional culture methods. We describe the validation of a new Real-Time PCR method to detect E. coli O157:H7 in five food matrixes. The complete system consists of the MicroSEQ E. coli O157:H7 Detection Kit, sample preparation (two sample preparation methods, the PrepSEQ Nucleic Acid Extraction Kit and the PrepSEQ Rapid Spin Sample Preparation Kit, were validated), the Applied Biosystems 7500 Fast Real-Time PCR instrument, and RapidFinder Express software. The test method was compared to the U.S. Department of Agriculture Microbiology Laboratory Guidebook 5.04 reference method for detecting E. coli O157:H7 in 25 g and 375 g ground beef and beef trim, and to the ISO 16654 reference method for detecting E. coli O157:H7 in 25 g spinach, orange juice, and apple juice. The MicroSEQ E. coli O157:H7 Detection Kit showed equivalent detection compared to the corresponding reference method based on Mantel-Haenszel Chi-square statistics for all matrixes tested. An independent validation confirmed these findings on ground beef. The MicroSEQ kit detected all 51 E. coli O157:H7 strains tested and showed good discrimination against an exclusivity panel of 30 strains.

Evaluation of applied biosystems MicroSEQ real-time PCR system for detection of Listeria spp. in food and environmental samples.

A complete system for real-time PCR detection of Listeria species was validated in five food matrixes and five environmental surfaces, namely, hot dogs, roast beef, lox (smoked salmon), pasteurized whole cow's milk, dry infant formula, stainless steel, plastic cutting board, ceramic tile, rubber sheets, and sealed concrete. The system consists of the MicroSEQ Listeria spp. Detection Kit, two sample preparation kits (PrepSEQ Nucleic Acid Extraction Kit and PrepSEQ Rapid Spin Sample Preparation Kit), the Applied Biosystems 7500 Fast Real-Time PCR instrument, and the RapidFinderTM Express v1.1 Software for data analysis. The test method was compared to the ISO 11290-1 reference method using an unpaired study design. The MicroSEQ Listeria spp. Detection Kit and the ISO 11290-1 reference method showed equivalent detection based on Chi-square analysis for all matrixes except hot dogs. For hot dogs, the MicroSEQ method detected more positives than the reference method for the low- and high-level inoculations, with all of the presumptive positives confirmed by the reference method. An independent validation study confirmed these findings on lox and stainless steel surface. The MicroSEQ kit detected all 50 Listeria strains tested and none of the 31 nontarget bacteria strains.

Identification and characterization of novel Salmonella mobile elements involved in the dissemination of genes linked to virulence and transmission.

The genetic diversity represented by >2,500 different Salmonella serovars provides a yet largely uncharacterized reservoir of mobile elements that can contribute to the frequent emergence of new pathogenic strains of this important zoonotic pathogen. Currently, our understanding of Salmonella mobile elements is skewed by the fact that most studies have focused on highly virulent or common serovars. To gain a more global picture of mobile elements in Salmonella, we used prediction algorithms to screen for mobile elements in 16 sequenced Salmonella genomes representing serovars for which no prior genome scale mobile element data were available. From these results, selected mobile elements underwent further analyses in the form of validation studies, comparative analyses, and PCR-based population screens. Through this analysis we identified a novel plasmid that has two cointegrated replicons (IncI1-IncFIB); this plasmid type was found in four genomes representing different Salmonella serovars and contained a virulence gene array that had not been previously identified. A Salmonella Montevideo isolate contained an IncHI and an IncN2 plasmid, which both encoded antimicrobial resistance genes. We also identified two novel genomic islands (SGI2 and SGI3), and 42 prophages with mosaic architecture, seven of them harboring known virulence genes. Finally, we identified a novel integrative conjugative element (ICE) encoding a type IVb pilus operon in three non-typhoidal Salmonella serovars. Our analyses not only identified a considerable number of mobile elements that have not been previously reported in Salmonella, but also found evidence that these elements facilitate transfer of genes that were previously thought to be limited in their distribution among Salmonella serovars. The abundance of mobile elements encoding pathogenic properties may facilitate the emergence of strains with novel combinations of pathogenic traits.

Kikuchi-Fujimoto disease: a sheep in wolf's clothing.

INTRODUCTION: Kikuchi-Fujimoto disease (KFD) is a rare, self-limiting disorder that typically affects the cervical lymph nodes. Recognition of this condition is crucial, especially because it can easily be mistaken for tuberculosis, lymphoma, or even adenocarcinoma. Awareness of this disorder will help prevent misdiagnosis and inappropriate treatment. METHODS: From January 2006 to December 2008, 30 patients who underwent a biopsy of a cervical lymph node and proved histologically to have KFD were enrolled in this study. We studied clinical manifestations, laboratory results, treatment, and recurrence for each patient. Patients were followed up for a mean period of 2 years. RESULTS: There were 24 women and 6 men, with a mean age of 18 years. Two patients had a past history of tuberculosis. Major clinical symptoms and signs were fever (70%) and lymphadenopathy (100%). The affected cervical lymph nodes were most commonly located in the posterior cervical triangle. Unilateral and bilateral cervical lymph nodes were affected in 25 and 5 patients, respectively. The affected lymph nodes were most commonly less than 3 cm in size. Leukopenia was observed in 46.7%, and a raised erythrocyte sedimentation rate was seen in 56.7% of the cases. Treatment strategies included no medication, nonsteroidal antiinflammatory drugs (NSAIDs) alone, steroids alone, or a combination of NSAIDs and steroids. Ninety percent improved within 3 months, whereas one patient showed improvement only after 9 months of continued treatment. No recurrence has since been noted. CONCLUSION: KFD is a benign disease that masquerades as other more sinister diseases and can lead to unnecessary treatment-induced physiologic, psychological, and financial morbidity to the patient. Tissue diagnosis is necessary in all cases, and an effective communication between the surgeon and the pathologist is imperative in making an accurate diagnosis.