Bacterial CRISPR systems provide acquired immunity against invading nucleic acids by activating RNA-programmable RNases and DNases. Cas13a and Cas12a enzymes bound to CRISPR RNA (crRNA) recognize ...specific nucleic acid targets, initiating cleavage of the targets as well as non-target (trans) nucleic acids. Here, we examine the kinetics of single-turnover target and multi-turnover trans-nuclease activities of both enzymes. High-turnover, non-specific Cas13a trans-RNase activity is coupled to rapid binding of target RNA. By contrast, low-turnover Cas12a trans-nuclease activity is coupled to relatively slow cleavage of target DNA, selective for DNA over RNA, indifferent to base identity, and preferential for single-stranded substrates. Combining multiple crRNA increases detection sensitivity of targets, an approach we use to quantify pathogen DNA in samples from patients suspected of Buruli ulcer disease. Results reveal that these enzymes are kinetically adapted to play distinct roles in bacterial adaptive immunity and show how kinetic analysis can be applied to CRISPR-based diagnostics.
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•Cas13a HEPN trans-RNase activation is directly coupled to rapid target RNA binding•Cas12a RuvC trans-nuclease activity is coupled to slow target DNA cleavage•Individual crRNA generate widely varying levels of targeted trans-cleavage•Pooling multiple crRNA allows pathogen quantification without target amplification
Chemical reaction kinetics; Biochemistry; Molecular biology
Acute respiratory infections (ARI) account for 14.3% of new born mortality and 15.9% of death among children aged 1 to 5 in India, making them significant contributors to morbidity and mortality in ...children. However, the bulk of these fatalities would be avoided if mothers were aware of the symptoms and signs of infections so that prompt referrals could be made. The goal of the current study was to gauge mothers of children under the age of five's knowledge of ARI and examine how well an education programme did in raising that knowledge. The information was gathered from 60 mothers of young children under the age of five in different areas around Gujarat State in India. The pre-intervention-post-test methodology was used. The mean pre-test observation score was 8.51, the mean post-test score was 19.31, and the mean difference was 11. There were 60 samples; the pre-test and post-test scores had standard deviations of 4.59 and 2.54, respectively. The calculated "t" value was also 17.29, the DF was 59, and the table value of "t" was 1.67. These findings demonstrate that the educational programme on ARI was successful in increasing the mother's knowledge.
The lower Goodlands Member of the Turtle Mountain Formation is exposed in a streambank outcrop on the western flank of Turtle Mountain, south-western Manitoba, Canada (49°0'2"N, 100°14'51"W). This ...outcrop was sampled for a 1.5 m section of microfossil-rich non-marine clay and coal-rich sediments deposited in a coastal plain environment during the early Palaeocene. These sediments were deposited 65.4-65 Ma and thus offer an opportunity to reconstruct terrestrial palaeoecology 0.6 to 1 Ma after the Cretaceous-Paleogene boundary (K-Pg) extinction event. We use quantitative palynology to reconstruct terrestrial plant ecology and explore ecological patterns of recovery and succession of these communities on a millennial time scale. Quantitative palynological analyses shows that at the time of deposition of the Goodlands Member the landscape on the coastal plain of the Western Interior Seaway - in what is now south-western Manitoba - was covered in a forested canopied swamp with tall standing swamp cypress and other conifers, Juglandaceae (walnut family), birch, alder, elms and other angiosperms, with an understory of ferns and ground cover of Sphagnum moss. Notably, palms were present but scarce, indicating a relatively warm climate. No successional pattern of diversity and community composition is evident in the samples; all samples showed high plant diversity (33-54 taxa/sample, H' 2-3). Vegetation in Manitoba recovered within 0.6 to 1 Ma following the K-Pg extinction event or was predominantly influenced by local environmental patterns independent of time.
The genitourinary syndrome of menopause (GSM) encompasses a range of symptoms linked to the genitourinary tract stemming from the reduction in estrogen levels following menopause. These symptoms may ...endure throughout a woman's lifetime. Platelet-rich plasma (PRP), known for its capacity to induce angiogenesis and the restoration effects of growth factors, has been widely employed in various disorders, including GSM. This article aims to comprehensively review the existing literature on the utilization of PRP for managing GSM. The search was executed in electronic databases, specifically PubMed, Scopus, and Google Scholar, up until April 2023. Eligible studies were meticulously chosen for inclusion in this systematic review. PRP emerges as a viable alternative for addressing vaginal atrophy, exhibiting favorable outcomes. Notably, it can be considered for patients with contraindications to hormonal therapy. However, the available body of evidence supporting the use of PRP for GSM remains limited. PRP presents itself as a promising agent, offering a patient-friendly, cost-effective alternative modality. To establish the efficacy of PRP in treating GSM definitively, future randomized trials are imperative.
Psoriasis (PSO) is a skin disorder with systemic inflammation and high coronary artery disease risk. A distinct lipid phenotype occurs in psoriasis, which is characterized by high plasma ...triglycerides (TGs) with typically normal or even low LDL-C. The extent to which cholesterol on LDL subfractions, such as small dense LDL-C (sdLDL-C), are associated with vulnerable coronary plaque characteristics in PSO remains elusive.
A recently developed equation for estimating sdLDL-C from the standard lipid panel was utilized in a PSO cohort (n = 200) with 4-year follow-up of 75 subjects. Coronary plaque burden was assessed by quantitative coronary computed tomography angiography (CCTA). Multivariate regression analyses were used for establishing associations and prognostic value of estimated sdLDL-C.
Estimated sdLDL-C was positively associated with non-calcified burden (NCB) and fibro-fatty burden (FFB), which remained significant after multivariate adjustment for NCB (β = 0.37; P = 0.050) and LDL-C adjustment for FFB (β = 0.29; P < 0.0001). Of note, total LDL-C calculated by the Friedewald equation was not able to capture these associations in the study cohort. Moreover, in the regression modelling estimated sdLDL-C was significantly predicting necrotic burden progression over 4 years follow-up (P = 0.015), whereas LDL-C did not. Finally, small LDL particles (S-LDLP) and small HDL particles (S-HDLP), along with large and medium TG-rich lipoproteins (TRLPs) had the most significant positive correlation with estimated sdLDL-C.
Estimated sdLDL-C has a stronger association than LDL-C with high-risk features of coronary atherosclerotic plaques in psoriasis patients.
URL: https://www.
gov . Unique identifiers: NCT01778569.
Background
Statin treatment is a potent lipid‐lowering therapy associated with decreased cardiovascular risk and mortality. Recent studies including the PARADIGM trial have demonstrated the impact of ...statins on promoting calcified coronary plaque.
Hypothesis
The degree of systemic inflammation impacts the amount of increase in coronary plaque calcification over 2 years of statin treatment.
Methods
A subgroup of 142 participants was analyzed from the Risk Stratification with Image Guidance of HMG CoA Reductase Inhibitor Therapy (RIGHT) study (NCT01212900), who were on statin treatment and underwent cardiac computed tomography angiography (CCTA) at baseline and 2‐year follow‐up. This cohort was stratified by baseline median levels of high‐sensitivity hs‐CRP and analyzed with linear regressions using Stata‐17 (StataCorp).
Results
In the high versus low hs‐CRP group, patients with higher baseline median hs‐CRP had increased BMI (median IQR; 29 27–31 vs. 27 24–28; p < .001), hypertension (59% vs. 41%; p = .03), and LDL‐C levels (97 77–113 vs. 87 75–97 mg/dl; p = .01). After 2 years of statin treatment, the high hs‐CRP group had significant increase in dense‐calcified coronary burden versus the low hs‐CRP group (1.27 vs. 0.32 mm2 100×; p = .02), beyond adjustment (β = .2; p = .03).
Conclusions
Statin treatment over 2 years associated with a significant increase in coronary calcification in patients with higher systemic inflammation, as measured by hs‐CRP. These findings suggest that systemic inflammation plays a role in coronary calcification and further studies should be performed to better elucidate these findings.
A combination of Ofloxacin, Tinidazole, Terbinafine hydrochloride, and Clobetasol propionate is used for treatment, control, prevention, and improvement of bacterial and fungal skin infections. Two ...chromatographic methods (HPLC and HPTLC) were developed for the quantitative determination of these four drugs in their combined topical formulation. The HPLC method employed a C
18
(250 × 4.6 mm, 5 μ) column as a stationary phase and acetonitrile: 0.1% triethylamine (pH 3.5) with gradient elution as mobile phase. The chromatographic separation in HPTLC was carried out on precoated silica 60F
254
plates using toluene:ethyl acetate:methanol:triethylamine:formic acid (5:4:1:0.3:0.3 v/v/v/v/v) as mobile phase. Both methods used 238 nm as detection wavelength and were able to separate all four drugs with resolved and sharp peaks. Developed methods were validated as per ICH Q2(R1) guidelines. Both methods were found to be linear in selected ranges (r
2
> 0.998), precise (RSD < 2%), accurate (recovery > 99.5%), sensitive and specific for quantitation of all four drugs. Validated methods were successfully applied for the quantitative determination of four drugs in the cream formulation and assay results were found to be in good agreement with the stated amount of each drug. Assay results were statistically compared (t-test) and no significant difference was observed (p > 0.05) between the results obtained by both methods.
We present a rare double aortic arch (DAA) diagnosis incidentally on CT in a 60-year-old male who presented with pneumonia. DAA is a vascular ring that typically manifests in infants or children due ...to compression of the esophagus or trachea, resulting in dysphagia or dyspnea. Diagnosis of DAA in adulthood is usually due to the delayed emergence of obstructive symptoms. We present a case of DAA in an adult patient without dysphagia or dyspnea. We discuss factors that can lead to the presentation of DAA in adults. These include an absence of associated congenital disabilities, insufficient tracheal or esophageal constriction in childhood and the onset of compressive symptoms later in life from decreased vascular compliance.
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•Ester functionalized surface active ionic liquids were synthesized.•Biodegradable and biocompatible catanionic vesicles were prepared.•Analytical techniques were used to study the ...structure of catanionic vesicles.•Small angle neutron scattering was used to characterize the aggregates.•Vesicles used to encapsulate curcumin.
Aqueous solubility and stability of curcumin could be ameliorated through its encapculation within the micellar and vesicular assemblies. This could protect curcumin from the attack of inactivating enzymes in the biological fluid and improve its medicinal and therapeutic effects. In our efforts to design surfactant based organized assemblies as drug carrier, herein, we had reported the formation of catanionic vesicles to encapsulate curcumin. The catanionic vesicles are formed through synergistic interaction between sodium butyrate (NaBut) and ester functionalized morpholinium and imidazolium-based surface active ionic liquids (SAILs). Fluorescence resonance energy transfer (FRET) and small-angle neutron scattering (SANS) were among the methods that were used to confirm the presence of curcumin within the bilayer of the vesicles. Loading capacity and stability of the curcumin within the micelles and vesicles were investigated. Results concludes that vesicles because of their higher hydrophobic character improve the encapsulation and inhibit the degradation of curcumin. The curcumin loaded catanionic vesicles were found effective against Gram-positive and Gram-negative bacteria and must be exploited for clinical applications. The newly fabricated system would pave the way for the stabilization of the curcumin and increased its application potential in the field of biotechnology and sustained drug delivery.
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