Simultaneous Kinetic Spectrophotometric Determination of Hydrazine and Isoniazid Using H-Point Standard Addition Method and Partial Least Squares Regression in Micellar Media

The present study describes the application of simultaneous kinetic spectrophotometric determination of hydrazine (HZ) and isoniazid (INH), using H-point standard addition method (HPSAM) and partial least squares (PLS) calibration. The methods are based on the difference observed in the rate of iron (III) reduction with HZ and INH, in the presence of 2,2\u27-bipyridine (Bpy) and the subsequent complex formation between the resulted Fe2+ and Bpy in a solution containing sodium dodecyl sulfate (SDS) as a micellar medium. INH and HZ can simultaneously be determined between the range of 0.08−6.0 and 1.0−80.0 μg mL−1, respectively. The results have shown that by the application of HPSAM, the simultaneous determination could be performed with the ratio of 1:1000 to 1:12.5 for INH-HZ. Through the HPSAM analysis, the relative standard deviations of HZ and INH were 2.5 and 1.2, respectively. The total relative standard error for applying the PLS method to 9 synthetic samples, in the concentration ranges of 0.0−20.0 μg mL−1 of HZ and 0.5−3.0 μg mL−1 of INH, was 3.19. Both proposed methods (PLS and HPSAM) were successfully applied to the simultaneous determination of HZ and INH in several commercially available isoniazid formulations and satisfactory results were obtained.</p

Genetic characterization of the central variable region in African swine fever virus isolates in the Russian federation from 2013 to 2017

African swine fever virus (ASFV), classified as genotype II, was introduced into Georgia in 2007, and from there, it spread quickly and extensively across the Caucasus to Russia, Europe and Asia. The molecular epidemiology and evolution of these isolates are predominantly investigated by means of phylogenetic analysis based on complete genome sequences. Since this is a costly and timeconsuming endeavor, short genomic regions containing informative polymorphisms are pursued and utilized instead. In this study, sequences of the central variable region (CVR) located within the B602L gene were determined for 55 ASFV isolates submitted from 526 active African swine fever (ASF) outbreaks occurring in 23 different regions across the Russian Federation (RF) between 2013 and 2017. The new sequences were compared to previously published data available from Genbank, representing isolates from Europe and Asia. The sequences clustered into six distinct groups

A guide to molecular characterization of genotype ii African swine fever virus: Essential and alternative genome markers

African swine fever is a contagious viral disease that has been spreading through Europe and Asia since its initial report from Georgia in 2007. Due to the large genome size of the causative agent, the African swine fever virus (ASFV), the molecular epidemiology, and virus evolution are analyzed by employing different markers. Most of these markers originate from single nucleotide polymorphisms or disparities in the copy number of tandem repeat sequences observed during the comparisons of full genome sequences produced from ASFVs isolated during different outbreaks. Therefore, consistent complete genome sequencing and comparative analysis of the sequence data are important to add innovative genomic markers that contribute to the delineation of ASFV phylogeny and molecular epidemiology during active circulation in the field. In this study, the molecular markers currently employed to assess the genotype II ASFVs circulating in Europe and Asia have been outlined

Whole-genome sequencing of African swine fever virus from wild boars in the Kaliningrad region reveals unique and distinguishing genomic mutations

Since the first report of outbreaks of African swine fever (ASF) in Georgia in 2007, the disease has expanded into Europe, Russia, and Asia, spreading rapidly via contact with infected animals including domestic pigs and wild boars. The vast expansion of this Genotype II African swine fever virus (ASFV) across wide-ranging territories and hosts inevitably led to the acquisition of novel mutations. These mutations could be used to track the molecular epidemiology of ASFV, provided that they are unique to strains restricted within a certain area. Whilst whole-genome sequencing remains the gold standard for examining evolutionary changes, sequencing of a single locus with significant variation and resolution power could be used as a rapid and cost-eective alternative to characterize multiple isolates from a single or related outbrea

Electrochemical sensor based on multi-walled carbon nanotubes and 4-(((4-mercaptophenyl)imino)methyl) benzene-1,2-diol for simultaneous determination of epinephrine in the presence of acetaminophen

A carbon paste electrode modified with 4-(((4-mercaptophenyl)imino)methyl)benzene-1,2-diol (MIB) and multi-walled carbon nanotubes MIB /CNT/CPE) was prepared for determination of epinefrine (EP) in the presence of acetaminophen (AC). Cyclic voltammetry, chronoamperometry and differential pulse voltammetry (DPV) techniques were used to investigate the modified electrode for the electrocatalytic oxidation of (EP) and (AC) in aqueous solutions. The separation of the oxidation peak potential for EP- AC was 200 mV. Under the optimum conditions, the calibration curve for EP was obtained in the range of 1.0 to 25.0 µM and 25.0 to 500.0 µM. The diffusion coefficient for the oxidation of EP at the surface of modified electrode was calculated as 5.76×10-5 cm2s-1.</p

Simultaneous Kinetic Spectrophotometric Determination of Hydrazine and Isoniazid Using H-Point Standard Addition Method and Partial Least Squares Regression in Micellar Media

The present study describes the application of simultaneous kinetic spectrophotometric determination of hydrazine (HZ) and isoniazid (INH), using H-point standard addition method (HPSAM) and partial least squares (PLS) calibration. The methods are based on the difference observed in the rate of iron (III) reduction with HZ and INH, in the presence of 2,2\u27-bipyridine (Bpy) and the subsequent complex formation between the resulted Fe2+ and Bpy in a solution containing sodium dodecyl sulfate (SDS) as a micellar medium. INH and HZ can simultaneously be determined between the range of 0.08−6.0 and 1.0−80.0 μg mL−1, respectively. The results have shown that by the application of HPSAM, the simultaneous determination could be performed with the ratio of 1:1000 to 1:12.5 for INH-HZ. Through the HPSAM analysis, the relative standard deviations of HZ and INH were 2.5 and 1.2, respectively. The total relative standard error for applying the PLS method to 9 synthetic samples, in the concentration ranges of 0.0−20.0 μg mL−1 of HZ and 0.5−3.0 μg mL−1 of INH, was 3.19. Both proposed methods (PLS and HPSAM) were successfully applied to the simultaneous determination of HZ and INH in several commercially available isoniazid formulations and satisfactory results were obtained.</p

Post-genomic era in agriculture and veterinary science: successful and proposed application of genetic targeting technologies

Gene editing tools have become an indispensable part of research into the fundamental aspects of cell biology. With a vast body of literature having been generated based on next generation sequencing technologies, keeping track of this ever-growing body of information remains challenging. This necessitates the translation of genomic data into tangible applications. In order to address this objective, the generated Next Generation Sequencing (NGS) data forms the basis for targeted genome editing strategies, employing known enzymes of various cellular machinery, in generating organisms with specifically selected phenotypes. This review focuses primarily on CRISPR/Cas9 technology in the context of its advantages over Zinc finger proteins (ZNF) and Transcription activator-like effector nucleases (TALEN) and meganucleases mutagenesis strategies, for use in agricultural and veterinary applications. This review will describe the application of CRISPR/Cas9 in creating modified organisms with custom-made properties, without the undesired non-targeted effects associated with virus vector vaccines and bioactive molecules produced in bacterial systems. Examples of the successful and unsuccessful applications of this technology to plants, animals and microorganisms are provided, as well as an in-depth look into possible future trends and applications in vaccine development, disease resistance and enhanced phenotypic traits will be discussed

African swine fever in the Primorsky Krai: disease situation and molecular and biological properties of the isolate recovered from a wild boar long bone

It is necessary to continue the analysis of the situation and molecular and biological properties of the current African swine fever virus isolates, recovered in the Russian border territories to cover the following tasks: eradication of African swine fever; development of effective disease surveillance and control programs; search for promising genome markers for the vaccine development; implementation of the differentiation strategy between vaccinated and non-vaccinated animals; and clustering of the isolates. The post-hoc analysis of some ASF epidemiological data and comparative genetic analysis of isolates circulating in the Far East Federal District suggested the agent introduction and spread routes, as well as the seasonality of the infection occurrence in the Primorsky Krai. It was established, that two ASFV subgenotypes (IGR-I и IGR-II), differentiated by intergenic region I73R/I329L, circulated in the region under study during the first months post infection. Analysis of biological properties of ASFV/Primorsky 19/WB-6723 isolate recovered from the long bone of a dead wild boar in the Primorsky Krai suggested that the isolate is highly virulent, able to cause peracute to subacute disease and up to 100% mortality among infected animals. The incubation period and duration of the disease course in experimentally infected pigs were 4–6 and 3–5 days post infection, respectively. The ASFV genome was detected in blood samples collected from infected pigs on 5–8 days post infection by real-time polymerase chain reaction. Specific antibodies in blood samples were not detected. The need in further research of molecular and biological properties of current ASFV isolates was reaffirmed. To prevent the continuation of the epizooty and deterioration of the current situation the approaches to the disease surveillance and control need to be modified

Accessibility to biologics and its impact on disease activity and quality of life in patients with rheumatoid arthritis in Kuwait

Objective: Biologics are indicated in rheumatoid arthritis (RA) in case of persistent high disease activity despite conventional disease-modifying anti-rheumatic drugs (cDMARDs) or patients with contraindications to cDMARDs or poor prognostic factors. The purpose of this study was to compare the prescription rates of biologics in Kuwaiti and non-Kuwaiti patients and to assess whether this had an impact on disease activity and quality of life in RA patients. Methods: Data were extracted from the Kuwait Registry for Rheumatic Diseases. Adult patients who satisfied the ACR classification criteria for RA from four major hospitals in Kuwait were evaluated from February 2013 through May 2018. The treatment agents, disease activity, and quality of life of Kuwaiti patients were compared with non-Kuwaiti patients. Results: A total of 1651 RA patients were included; 806 (48.8%) were Kuwaiti patients. Among Kuwaiti patients, 62.5% were on biologic drugs in comparison with 14% of non-Kuwaiti patients. In comparison with non-Kuwaiti patients, Kuwaiti patients had significantly lower numbers of swollen joints (p < 0.001) and disease activity score-28 scores (p = 0.02) and less steroid use (p < 0.001) yet a significantly higher health assessment questionnaire-disability index (p < 0.001). Regression analysis showed that DAS-28 scores were significantly associated with the treatment type (p < 0.001) and that nationality was significantly predictive of the treatment type (p < 0.001). Conclusion: In the setting of easy accessibility to treatment for Kuwaiti patients, biologics were prescribed by rheumatologists at a higher rate than for non-Kuwaitis. This may explain the lower disease activity and the lower rate of steroid use in Kuwaiti patients than non-Kuwaitis. Key points: • Significant discrepancies in the rates of prescribing biologic therapies between KP and NKP in Kuwait were observed. • Several treatment outcomes were significantly better in the KP group than in the NKP group even after adjustment of confounding factors. • The poor access to biologic therapies was suggested to limit the effectiveness of RA treatments in the NKP group

Whole-genome sequencing of African swine fever virus from wild boars in the Kaliningrad region reveals unique and distinguishing genomic mutations

IntroductionSince the first report of outbreaks of African swine fever (ASF) in Georgia in 2007, the disease has expanded into Europe, Russia, and Asia, spreading rapidly via contact with infected animals including domestic pigs and wild boars. The vast expansion of this Genotype II African swine fever virus (ASFV) across wide-ranging territories and hosts inevitably led to the acquisition of novel mutations. These mutations could be used to track the molecular epidemiology of ASFV, provided that they are unique to strains restricted within a certain area. Whilst whole-genome sequencing remains the gold standard for examining evolutionary changes, sequencing of a single locus with significant variation and resolution power could be used as a rapid and cost-effective alternative to characterize multiple isolates from a single or related outbreak.Material and methodsASFVs obtained during active ASF outbreaks in the Russian region of Kaliningrad between 2017 and 2019 were examined. Since all of the viruses belonged to Genotype II and no clear differentiation based on central variable region (CVR) sequencing was observed, the whole-genome sequences of nine ASFV isolates from this region were determined. To obtain insights into the molecular evolution of these isolates, their sequences were compared to isolates from Europe, Asia, and Africa.ResultsPhylogenetic analysis based on the whole-genome sequences clustered the new isolates as a sister lineage to isolates from Poland and Germany. This suggests a possible shared origin followed by the addition of novel mutations restricted to isolates from this region. This status as a sister lineage was mirrored when analyzing polymorphisms in MGF-505-5R and MGF-110-7L, whilst a polymorphism unique to sequences from Kaliningrad was identified at locus K145R. This newly identified mutation was able to distinguish the isolates obtained from Kaliningrad with sequences of Genotype II ASFVs available on GenBank.DiscussionThe findings of this study suggest that ASFVs circulating in Kaliningrad have recently obtained this mutation providing an additional marker to the mutations previously described