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Cureus Apr 202390% of visually impaired people live in developing countries. There are various types of vision impairment, but the focus of the current study is retinitis pigmentosa...
INTRODUCTION
90% of visually impaired people live in developing countries. There are various types of vision impairment, but the focus of the current study is retinitis pigmentosa (RP). Up to now, 150 mutations have been reported that are linked with RP.
METHODOLOGY
Healthy and affected members from two Pakistani families (RP01 and RP02) segregating autosomal recessive RP were selected for DNA extraction. PCR was conducted, and the amplified PCR products were analyzed using Polyacrylamide Gel Electrophoresis (PAGE) and visualized in the Gel Doc system for linkage analysis. The Gene Hunter 2.1r5 tool in the Simple Linkage v5.052 beta software suite was used to conduct multipoint parametric linkage analysis on the two consanguineous families examined on the 6K Illumina array. Exons and intron-exon borders of all known arRP genes found in homozygous areas were sequenced in the matching probands using a 3130 automated sequencer and the Big Dye Terminator Cycle Sequencing Kit v3.1. The mutation study was carried out using the AlaMut 1.5 program.
RESULTS
In both families, linkage analysis was performed using microsatellite marker DIS422 for gene and microsatellite marker D8S2332 for gene . Multipoint linkage analysis identifies genomic regions that could potentially contain the genetic defect. In family RP01, only a single peak with a maximal multipoint LOD score of 3.00 was identified on chromosome 1, whereas in family RP02, multiple peaks with multipoint LOD scores of 1.80 were identified on chromosome 8. Analysis of the gene revealed a homozygous substitution of glycine for valine (c.1152T>G; p.V243G), whereas the gene demonstrated that leucine was substituted for proline as a result of cytosine to thymine transfer (c.3419C>T; p. P1035L). Conclusion: Homozygosity mapping is a powerful method for finding genetic abnormalities that are both precise and comprehensive for identifying harmful variations in consanguineous families. This method is invaluable for providing accurate clinical diagnostic and genetic advice in remote regions of Pakistan while also increasing knowledge about autosomal recessive diseases and the dangers of mixing.
PubMed: 37267051
DOI: 10.7759/cureus.37933 -
Annals of Work Exposures and Health Aug 2021Exposure to asbestos fibres is linked to numerous adverse health effects and the use of asbestos is currently banned in many countries. Still, asbestos applications are...
Exposure to asbestos fibres is linked to numerous adverse health effects and the use of asbestos is currently banned in many countries. Still, asbestos applications are present in numerous residential and professional/industrial buildings or installations which need to be removed. Exposure measurements give good insight in exposure levels on the basis of which the required control regime is determined to ensure that workers are protected against adverse health effects. However, it is a costly and time-consuming process to measure all situations as working conditions and materials may vary greatly. Therefore, the mechanistic model 'Asbestos Removal Exposure Assessment Tool (AREAT)' was developed to estimate exposure to respirable asbestos fibres released during asbestos abatement processes where measurements are not available. In such instances tailored control regimes can be implemented based on modelled exposure levels. The mechanistic model was developed using scientific literature, an in-house asbestos abatement dataset, and knowledge with regard to previously developed models. Several exposure determinants such as the substance emission potential, activity emission potential, control measures, and dilution in air were identified and specific modifiers were developed for each category. Through an algorithm, AREAT calculates a dimensionless score based on the model inputs. The model was calibrated using a statistical model on an extensive measurement dataset containing a broad variety of exposure scenarios. This statistical model enabled the translation of dimensionless AREAT scores to actual estimated fibre concentrations in fibres m-3. In total, 370 personal inhalation exposure measurements from 71 different studies were used for calibration of AREAT. Of these measurements, in 191 cases (52%) with microscopic analysis (all asbestos fibre analyses were conducted with scanning electron microscopy/energy dispersive X-ray analysis in accordance with ISO 14966) no fibres were detected and the limit of detection values(LODs) were given. To assess the influence of the large number of measurements with exposures below LOD values on the performance of the model, calibrations were performed on the total dataset and the selection of data excluding measurements below LOD. The AREAT model correlated well with the datasets, with a Pearson correlation of 0.73 and 0.8 and Spearman rank correlation of 0.56 and 0.8. The model was fitted to estimate a typical exposure value [i.e. geometric mean (GM) exposures], but it is recommended to use a more conservative worst case higher percentile (for example the 90th percentile; which adds a factor of 17.3 based on the model uncertainty on the GM estimate), to account for variability in the measurements and uncertainty in model estimates. This work has shown the development and calibration of a mechanistic model, capable of estimating asbestos fibre exposures during asbestos abatement processes. The AREAT model will be implemented as a lower tier exposure model in a risk assessment tool used within the Netherlands to plan abatement processes and to develop control strategies.
Topics: Air Pollutants, Occupational; Asbestos; Calibration; Humans; Inhalation Exposure; Occupational Exposure
PubMed: 33791749
DOI: 10.1093/annweh/wxaa112 -
Current Biology : CB Dec 2021Many aspects of sleep are heritable, but only a few sleep-regulating genes have been reported. Here, we leverage mouse models to identify and confirm a previously...
Many aspects of sleep are heritable, but only a few sleep-regulating genes have been reported. Here, we leverage mouse models to identify and confirm a previously unreported gene affecting sleep duration-dihydropyrimidine dehydrogenase (Dpyd). Using activity patterns to quantify sleep in 325 Diversity Outbred (DO) mice-a population with high genetic and phenotypic heterogeneity-a linkage peak for total sleep in the active lights off period was identified on chromosome 3 (LOD score = 7.14). Mice with the PWK/PhJ ancestral haplotype at this location demonstrated markedly reduced sleep. Among the genes within the linkage region, available RNA sequencing data in an independent sample of DO mice supported a highly significant expression quantitative trait locus for Dpyd, wherein reduced expression was associated with the PWK/PhJ allele. Validation studies were performed using activity monitoring and EEG/EMG recording in Collaborative Cross mouse strains with and without the PWK/PhJ haplotype at this location, as well as EEG and EMG recording of sleep and wake in Dpyd knockout mice and wild-type littermate controls. Mice lacking Dpyd had 78.4 min less sleep during the lights-off period than wild-type mice (p = 0.007; Cohen's d = -0.94). There was no difference in other measured behaviors in knockout mice, including assays evaluating cognitive-, social-, and affective-disorder-related behaviors. Dpyd encodes the rate-limiting enzyme in the metabolic pathway that catabolizes uracil and thymidine to β-alanine, an inhibitory neurotransmitter. Thus, data support β-alanine as a neurotransmitter that promotes sleep in mice.
Topics: Animals; Dihydrouracil Dehydrogenase (NADP); Haplotypes; Mice; Mice, Knockout; Sleep; beta-Alanine
PubMed: 34653361
DOI: 10.1016/j.cub.2021.09.049 -
Prognostic value of the H FPEF score in patients undergoing transcatheter aortic valve implantation.ESC Heart Failure Feb 2021The aim of this study was to assess the prognostic value of the H FPEF score in patients undergoing transcatheter aortic valve implantation (TAVI) for severe aortic...
AIMS
The aim of this study was to assess the prognostic value of the H FPEF score in patients undergoing transcatheter aortic valve implantation (TAVI) for severe aortic stenosis (AS) and preserved left ventricular ejection fraction (EF).
METHODS AND RESULTS
In this multicentre study, a total of 832 patients from two German high-volume centres, who received TAVI for severe AS and preserved EF (≥50%), were identified for calculation of the H FPEF score. Patients were dichotomized according to low (0-5 points; n = 570) and high (6-9 points; n = 262) H FPEF scores. Kaplan-Meier and Cox regression analyses were applied to assess the prognostic impact of the H FPEF score. We observed a decrease in stroke volume index (-2.04 mL/m /point) and mean transvalvular gradients (-1.14 mmHg/point) with increasing H FPEF score translating into a higher prevalence of paradoxical low-flow, low-gradient AS among patients with high H FPEF score. One year after TAVI, the rates of all-cause (low vs. high H FPEF score: 8.0% vs. 19.4%, P < 0.0001) and cardiovascular (CV) mortality (1.9% vs. 9.0%, P < 0.0001) as well as the rate of CV mortality or rehospitalization for congestive heart failure (6.4% vs. 23.2%, P < 0.0001) were higher in patients with high H FPEF score compared with those with low H FPEF score. After multivariable analysis, a high H FPEF score remained independently predictive of all-cause mortality [hazard ratio 1.59 (1.28-2.35), P = 0.018] and CV mortality or rehospitalization for congestive heart failure [hazard ratio 2.92 (1.65-5.15), P < 0.001]. Among the H FPEF score variables, atrial fibrillation, pulmonary hypertension, and elevated left ventricular filling pressure were the strongest outcome predictors.
CONCLUSIONS
The H FPEF score serves as an independent predictor of adverse CV and heart failure outcome among TAVI patients with preserved EF. A high H FPEF score is associated with the presence of paradoxical low-flow, low-gradient AS, the HFpEF in patients with AS. By identifying patients in advanced stages of HFpEF, the H FPEF score might be useful as a risk prediction tool in patients with preserved EF scheduled for TAVI.
Topics: Aortic Valve Stenosis; Heart Failure; Humans; Prognosis; Stroke Volume; Transcatheter Aortic Valve Replacement; Ventricular Function, Left
PubMed: 33215870
DOI: 10.1002/ehf2.13096 -
Experimental and Therapeutic Medicine Apr 2020Developmental dysplasia of the hip (DDH), previously known as congenital hip dislocation, is a frequently disabling condition characterized by premature arthritis later...
Developmental dysplasia of the hip (DDH), previously known as congenital hip dislocation, is a frequently disabling condition characterized by premature arthritis later in life. Genetic factors play a key role in the aetiology of DDH. In the present study, a genome-wide linkage scan with the Affymetrix 10K GeneChip was performed on a four-generation Chinese family, which included 19 healthy members and 5 patients. Parametric and non-parametric multipoint linkage analyses were carried out with Genespring GT v.2.0 software, and the logarithm of odds (LOD) score and nonparametric linkage (NPL) score were calculated. Parametric linkage analysis was performed, assuming an autosomal recessive trait with full penetrance and Affymetrix 'Asian' allele frequencies. The strongest evidence for linkage was found on chromosome 8q23-24, with a peak LOD score of 2.658 (θ=0), covering 2.377 Mb from single nucleotide polymorphisms (SNPs) rs724717 to rs720132. This interval included nine additional successive SNPs: rs1566071, rs1902121, rs756404, rs702768, rs777813, rs2033995, rs147959, rs2884367 and rs1898287. The same region also yielded the highest NPL score of 2.883 (P=0.0156) from the non-parametric multipoint linkage analysis. Additionally, the second highest NPL score of 2.727 (P=0.0156) and LOD score of 2.528 (θ=0) were obtained on chromosome 12p12 for three consecutive markers (rs1919980, rs763853 and rs725124). This region overlapped a narrow distance of 0.642 Mb. Notably, in addition to these two regions; no significant linkage was identified for other chromosomal regions (with LOD and NPL scores >2.0). For the first time, at least for this pedigree, the evidence in the present study showed that DDH is mapped to two novel regions at 8q23-q24 and 12p12.
PubMed: 32256763
DOI: 10.3892/etm.2020.8513 -
Frontiers in Immunology 2022We identified and genes as putative effectors of the Quantitative Trait locus (QTL) that we mapped at distal chromosome 7 named for Inflammatory response modulator 1,...
We identified and genes as putative effectors of the Quantitative Trait locus (QTL) that we mapped at distal chromosome 7 named for Inflammatory response modulator 1, controlling acute inflammatory response (AIR) and the production of IL-1β, dependent on the activation of the NLRP3 inflammasome. We obtained the mapping through genome-wide linkage analysis of Single Nucleotide Polymorphisms (SNPs) in a cross between High (AIRmax) and Low (AIRmin) responder mouse lines that we produced by several generations of bidirectional selection for Acute Inflammatory Response. A highly significant linkage signal (LOD score peak of 72) for IL-1β production limited a 4 Mbp interval to chromosome 7. Sequencing of the locus region revealed 14 SNPs between "High" and "Low" responders that narrowed the locus to a 420 Kb interval. Variants were detected in non-coding regions of , and genes and at the first exon of gene, resulting in an E19K substitution in the protein ASC (apoptosis associated speck-like protein containing a CARD) an adaptor molecule in the inflammasome complex. Silencing of inhibited IL1-β production by stimulated macrophages and the E19K ASC mutation carried by AIRmin mice impaired the IL-1β response and the formation of ASC specks in stimulated cells. IL-1β and ASC specks play major roles in inflammatory reactions and in inflammation-related diseases. Our results delineate a novel genetic factor and a molecular mechanism affecting the acute inflammatory response.
Topics: Animals; CARD Signaling Adaptor Proteins; Genetic Linkage; Inflammasomes; Inflammation; Mice; Quantitative Trait Loci
PubMed: 35799794
DOI: 10.3389/fimmu.2022.899569 -
International Journal of Molecular... Dec 2022In the current study, the reversed-phased high-pressure liquid chromatography (RP-HPLC) method was proposed for the estimation of lignocaine hydrochloride (LIG),...
In the current study, the reversed-phased high-pressure liquid chromatography (RP-HPLC) method was proposed for the estimation of lignocaine hydrochloride (LIG), hydrocortisone (HYD) and Ketoprofen (KET) according to International Conference for Harmonization (ICH) Q2 R1 guidelines, in a gel formulation. The chromatographic evaluation was executed using Shimadzu RP-HPLC, equipped with a C8 column and detected using UV at 254 nm wavelength, using acetonitrile and buffer (50:50) as a mobile phase and diluent, at flow rate 1 mL/min and n injection volume of 20 μL. The retention time for LIG, HYD, and KET were 1.54, 2.57, and 5.78 min, correspondingly. The resultant values of analytical recovery demonstrate accuracy and precision of the method and was found specific in identification of the drugs from dosage form and marketed products. The limit of detection (LOD) for LIG, HYD, and KET were calculated to be 0.563, 0.611, and 0.669 ppm, while the limit of quantification (LOQ) was estimated almost at 1.690, 1.833, and 0.223 ppm, respectively. The AGREE software was utilized to evaluate the greenness score of the proposed method, and it was found greener in score (0.76). This study concluded that the proposed method was simple, accurate, precise, robust, economical, reproducible, and suitable for the estimation of drugs in transdermal gels.
Topics: Ketoprofen; Chromatography, High Pressure Liquid; Hydrocortisone; Limit of Detection; Reproducibility of Results
PubMed: 36613881
DOI: 10.3390/ijms24010440 -
Plants (Basel, Switzerland) Sep 2022Soil salinity is a major abiotic stress that causes disastrous losses in crop yields. To identify favorable alleles that enhance the salinity resistance of rice ( L.)...
Soil salinity is a major abiotic stress that causes disastrous losses in crop yields. To identify favorable alleles that enhance the salinity resistance of rice ( L.) crops, a set of 120 Cheongcheong Nagdong double haploid (CNDH) lines derived from a cross between the variety Cheongcheong and the variety Nagdong were used. A total of 23 QTLs for 8 different traits related to salinity resistance on chromosomes 1-3 and 5-12 were identified at the seedling stage. A QTL related to the salt injury score (SIS), qSIS-3b, had an LOD score of six within the interval RM3525-RM15904 on chromosome 3, and a phenotypic variation of 31% was further examined for the candidate genes. Among all the CNDH populations, five resistant lines (CNDH 27, CNDH 34-1, CNDH 64, CNDH 78, and CNDH 112), five susceptible lines (CNDH 52-1, CNDH 67, CNDH 69, CNDH 109, and CNDH 110), and the parent lines Cheongcheong and Nagdong were selected for relative gene expression analysis. Among all the genes, two candidate genes were highly upregulated in resistant lines, including the auxin-responsive protein IAA13 (Os03g0742900) and the calmodulin-like protein 4 (Os03g0743500-1). The calmodulin-like protein 4 (Os03g0743500-1) showed a higher expression in all the resistant lines than in the susceptible lines and a high similarity with other species in sequence alignment and phylogenetic tree, and it also showed a protein-protein interaction with other important proteins. The genes identified in our study will provide new genetic resources for improving salt resistance in rice using molecular breeding strategies in the future.
PubMed: 36235331
DOI: 10.3390/plants11192467 -
Analytical Sciences : the International... Sep 2023The sensitive, non-destructive constant wavelength (CW) and constant energy (CE) SFS techniques have been used for the simultaneous determination of 1-amino pyrene (AP)...
The sensitive, non-destructive constant wavelength (CW) and constant energy (CE) SFS techniques have been used for the simultaneous determination of 1-amino pyrene (AP) and 1-napthyl amine (NA) in their mixtures without prior separation via optimization of different experimental conditions (Δλ 70.0 nm, Δν 4000.0 cm, scan rate 240.0 nm/min, 25.0 °C, methanol). Amplitude-concentration plots have been linear for 1-amino pyrene, AP (0.01-0.1 mg/L) and 1-napthyl amine, NA (0.1-1.0 mg/L). In aqueous methanolic binary mixtures, the mean recoveries (RSD, LOD and LOQ) of AP were found to be 100.09% (0.053, 0.008 mg/L and 0.034 mg/L) for emission, 100.11% (0.141, 0.008 mg/L, 0.034 mg/L) for CWSFS, 100.05% (0.109, 0.007 mg/L and 0.032 mg/L) for first derivative CWSFS, 100.00% (0.148, 0.007 mg/L and 0.031 mg/L) for CESFS, 99.99% (0.109, 0.008 mg/L and 0.035 mg/L) for first derivative CESFS modes respectively. Additionally, for NA the mean recoveries (RSD, LOD and LOQ) were 100.29% (0.360, 0.046 mg/L and 0.204 mg/L) for emission, 100.06% (0.089, 0.098 mg/L, 0.436 mg/L) for CWSFS, 100.09% (0.144, 0.065 mg/L and 0.288 mg/L) for first derivative CWSFS, 100.05% (0.178, 0.077 mg/L and 0.339 mg/L) for CESFS, 100.03% (0.181, 0.082 mg/L and 0.364 mg/L) for first derivative CESFS modes respectively. Considering their safety and greenness, these methods might be considered as green tools using analytical eco-scale approaches (eco-scale score 88.0).
PubMed: 37244980
DOI: 10.1007/s44211-023-00368-8 -
Kidney International Feb 2022Amyloid A amyloidosis is a serious clinical condition resulting from the systemic deposition of amyloid A originating from serum amyloid A proteins with the kidneys...
Amyloid A amyloidosis is a serious clinical condition resulting from the systemic deposition of amyloid A originating from serum amyloid A proteins with the kidneys being the most commonly and earliest affected organ. Previously described amyloid A amyloidosis is linked to increased production and deposition of serum amyloid A proteins secondary to inflammatory conditions arising from infectious, metabolic, or genetic causes. Here we describe a family with primary amyloid A amyloidosis due to a chr11:18287683 T>C (human genome version19) mutation in the SAA1 promoter linked to the amyloidogenic SAA1.1 haplotype. This condition leads to a doubling of the basal SAA1 promoter activity and sustained elevation of serum amyloid A levels that segregated in an autosomal dominant pattern in 12 genetically affected and in none of six genetically unaffected relatives, yielding a statistically significant logarithm of odds (LOD) score over 5. Affected individuals developed proteinuria, chronic kidney disease and systemic deposition of amyloid composed specifically of the SAA1.1 isoform. Tocilizumab (a monoclonal antibody against the interleukin-6 receptor) had a beneficial effect when prescribed early in the disease course. Idiopathic forms represent a significant and increasing proportion (15-20%) of all diagnosed cases of amyloid A amyloidosis. Thus, genetic screening of the SAA1 promoter should be pursued in individuals with amyloid A amyloidosis and no systemic inflammation, especially if there is a positive family history.
Topics: Amyloidosis; Humans; Mutation; Promoter Regions, Genetic; Serum Amyloid A Protein
PubMed: 34560138
DOI: 10.1016/j.kint.2021.09.007