Molecular Assessments of Antimicrobial Protein Enterocins and Quorum Sensing Genes and Their Role in Virulence of the Genus Enterococcus.

Enterococcus has emerged as an opportunistic pathogen because of its antibiotic resistance and virulence profile, which makes it a causative agent of several diseases like endocarditis, surgical site, and urinary tract infections. Currently, species of this genus are the 2nd most frequently isolated microorganisms from hospital-acquired infections. Significant association with hospitals and unhygienic conditions of the environments has made them resistant to a wide range of antibiotics. On the brighter side, enterococci have the ability to produce antimicrobial proteins (i.e., enterocins) that exhibit wide antagonistic activity, thus making them useful microbes in the food and pharmaceutical industries. Enterocins are also involved in niche control in gut microbiota which is regulated by the quorum sensing (QS) system. A bacterial communication system that is controlled by the fsr operon in enterococci consists of FsrABDC, ef1097, and GelE/SprE genes. Hence, the present study was conducted for molecular assessment of enterocins and quorum sensing genes, inter-environmental correlation, and species prevalence of enterococci isolated from different environmental niches of Karachi, Pakistan. Obtained results revealed the highest prevalence of E. faecium and E. faecalis in all environments. Bacterial antagonism and enterocin genes were observed significantly high in poultry environments. The inter-environmental correlation indicated a strong positive correlation of freshwater with sewage and soil environments. Similarly, the fsr regulatory system was mostly identified in poultry-related environments, and a significant correlation between QS system and biofilm formation was established. In conclusion, this study confirmed the high prevalence of E. faecium in all tested sources, high enterocin production in non-clinical environments, and more fsr regulatory genes in poultry-related environments.

Symmetrical Ligand's Fabricated Porous Silicon Surface Based Photoluminescence Sensor for Metal Detection and Entrapment.

This study was based on the development of surface-based photoluminescence sensor for metal detection, quantification, and sample purification employing the solid sensory chip having the capability of metal entrapment. The Co(II), Cu(II) and Hg(II) sensitive fluorescence sensor (TP) was first synthesized and characterized its sensing abilities towards tested metal ions by using fluorescence spectral investigation while the synthesis and complexation of the receptor was confirmed by the chromogenic, optical, spectroscopic and spectrometric analysis. Under optical investigation, the ligand solution exhibited substantial chromogenic changes as well as spectral variations upon reacting with copper, cobalt, and mercuric ions, while these behaviors were not seen for the rest of tested metallic ions i.e., Na+, Ag+, Ni2+, Mn2+, Pd2+, Pb2+, Cd2+, Zn2+, Sn2+, Fe2+, Fe3+, Cr3+, and Al3+. These colorimetric alterations and spectral shifting could potentially be employed to detect and quantify these specific metal ions. After the establishment of the ligand's selective complexation ability towards selected metals, it was fabricated over the substituted porous silicon surface (FPS) keeping in view of the development of surface-based photoluminescence sensor (TP-FPS) for the selected metal sensation and entrapment to purify the sample just be putting off the metal entrapped sensory solid chip. Surface characterization and ligand fabrication was inspected by plan and cross sectional electron microscopic investigations, vibrational and electronic spectral analysis. The sensitivity of the ligand (TP) in the solution phase metal discrimination was determined by employing the fluorescence titration analysis of the ligand solution after progressive induction of Co2+, Cu2+, and Hg2+, which afford the detection limit values of 2.14 × 10- 8, 3.47 × 10- 8 and 3.13 × 10- 3, respectively. Concurrently, photoluminescence titration of the surface fabricated sensor (TP-FPS) revealed detection limit values of 3.14 × 10- 9, 7.43 × 10- 9, and 8.21 × 10- 4, respectively, for the selected metal ions.

Decision algorithm for educational institute selection with spherical fuzzy heronian mean operators and Aczel-Alsina triangular norm.

This article presents a novel study of spherical fuzzy sets (SFSs), a more comprehensive framework of intuitionistic fuzzy sets and picture fuzzy sets. The SFS allows the decision-makers (DMs) to cope with complicated and insufficient information during the aggregation process. The Heronian mean (HrM) model theory is also utilized to express correlation among different input arguments or characteristics. Recently, the theory of Aczel Alsina triangular norms gained a lot of attention from various research scholars and has many capabilities to provide smooth approximations during decision analysis. In this article, we developed some appropriate operations of Aczel Alsina t-norms and t-conorms in light of spherical fuzzy (SF) information. We develop new mathematical ways to look at SF data to keep clarity and sufficient information. These are the SF Aczel Alsina Heronian mean (SFAAHrM) and SF Aczel Alsina weighted Heronian mean (SFAAWHrM) operators. Furthermore, we also present a list of new strategies based on Aczel Alsina operations, such as SF Aczel Alsina geometric Heronian mean (SFAAGHrM) and SF Aczel Alsina weighted geometric Heronian mean (SFAAWGHrM) operators. Some notable properties are also characterized to show the validity and effectiveness of our derived mathematical approaches. Considering our derived strategies, an algorithm for the multiple attribute decision-making (MADM) problem is established to resolve complicated real-life applications. A numerical example presents the compatibility of derived approaches and provides a solid mechanism to improve the performance of educational institutes. A comparison technique is also demonstrated to show the applicability and consistency of diagnosed approaches by contrasting the findings of pioneered approaches with existing methodologies.

Ratiometric Fluorescence and Chromogenic Probe for Trace Detection of Selected Transition Metals.

The design and development of a fluorescence sensor aimed at detecting and quantifying trace amounts of toxic transition metal ions within environmental, biological, and aquatic samples has garnered significant attention from diagnostic and testing laboratories, driven by the imperative to mitigate the health risks associated with these contaminants. In this context, we present the utilization of a heterocyclic symmetrical Schiff Base derivative for the purpose of fluorogenic and chromogenic detection of Co2+, Cu2+ and Hg2+ ions. The characterization of the ligand involved a comprehensive array of techniques, including physical assessments, optical analyses, NMR, FT-IR, and mass spectrometric examinations. The mechanism of ligand-metal complexation was elucidated through the utilization of photophysical parameters and FT-IR spectroscopic analysis, both before and after the interaction between the ligand and the metal salt solution. The pronounced alterations observed in absorption and fluorescence spectra, along with the distinctive chromogenic changes, following treatment with Co2+, Cu2+ and Hg2+, affirm the successful formation of complexes between the ligands and the treated metal ions. Notably, the receptor's complexation response exhibited selectivity towards Co(II), Cu(II), and Hg(II), with no observed chromogenic changes, spectral variations, or band shifts for the various tested metal ions, including Na+, Ag+, Ni2+, Mn2+, Pd2+, Pb2+, Cd2+, Zn2+, Sn2+, Fe2+, Fe3+, Cr3+ and Al3+. This absence of interaction between these metal ions and the ligand could be attributed to their compact or inadequately conducive conduction bands for complexation with the ligand's structural composition. To quantify the sensor's efficacy, fluorescence titration spectra were employed to determine the detection limits for Co2+, Cu2+ and Hg2+, yielding values of 2.92 × 10-8, 8.91 × 10-8, and 4.39 × 10-3 M, respectively. The Benesi-Hildebrand plots provided association constant values for the ligand-cobalt, ligand-copper, and ligand-mercury complexes as 0.74, 2.52, and 13.89 M-1, respectively.

Decision algorithm for picture fuzzy sets and Aczel Alsina aggregation operators based on unknown degree of wights.

Aggregation operators (AOs) are well-known and efficient mathematical tools that are utilized to overcome the impact of imprecise and vague information during the aggregation process. The theoretical concepts of Aczel Alsina aggregation expressions are an extension of triangular norms and become a hot research topic in the environment of the fuzzy framework. The power operators provide a smooth approximation and are used to mitigate the influence of redundant or insufficient information on the attributes or criteria. Some robust aggregation approaches are developed by combining two different theories, like power operators and Aczel Alsina aggregation tools. This article aims to explore the theory of picture fuzzy sets (PFSs), an extended version of fuzzy sets, and intuitionistic fuzzy sets. Some robust operations of Aczel Alsina aggregation tools are also present in light of the picture fuzzy environment. We established a class of new methodologies in the light of picture fuzzy information, including picture fuzzy Aczel Alsina power weighted average (PFAAPWA) and picture fuzzy Aczel Alsina power ordered weighted average (PFAAPOWA) operators. We also developed an appropriate approach like picture fuzzy Aczel Alsina power weighted geometric (PFAAPWG) and picture fuzzy Aczel Alsina power ordered weighted geometric (PFAAPOWG) operators. Notable properties and characteristics of proposed methodologies are also demonstrated. Our invented approaches not only aggregate complicated information but can clearly define interrelationships among several arguments. Moreover, we establish an algorithm for the multi-attribute group decision-making (MAGDM) problem to handle the impact of redundant and vague information on human opinions. Finally, we study an experimental case study to evaluate an appropriate optimal option from available options. To reveal consistency and effectiveness of developed approaches, influence study by changing various parametric values and comparative study by comparing results of existing approaches.

The implementation of an image-guided system at a proton therapy center facility.

PURPOSE: Pencil Beam Scanning (PBS) proton therapy has similar requirements on patient alignment to within 1 mm and 1-degree accuracy as photon radiosurgery. This study describes general workflow, acceptance, and commissioning test procedures and their respective results for an independent robotic arm used for Image Guided Radiotherapy (IGRT) for a Proton Therapy System. METHODS: The system is equipped with kV-imaging techniques capable of orthogonal and Cone-Beam Computed Tomography (CBCT) imaging modalities mounted on an independent robotic arm gantry attached to the ceiling. The imaging system is capable of 360-degree rotation around patients to produce CBCT and kilovoltage orthogonal images. The imaging hardware is controlled by Ehmet Health XIS software, and MIM Software handles the image fusion and registration to an acceptable accuracy of ≤1-mm shifts for patients' alignment. The system was tested according to the requirements outlined in the American Association of Physicists in Medicine (AAPM) Task Group (TG) 142 and TG 179. The system tests included (1) safety, functionality, and connectivity, (2) mechanical testing, (3) image quality, (4) image registration, and (5) imaging dose. Additional tests included imaging gantry isocentricity with a laser tracker and collision-avoiding system checks. RESULTS: The orthogonal and volumetric imaging are comparable in quality to other commercially available On-Board Imagers (OBI) systems. The resulting spatial resolution values were 1.8-, 0.8-, and 0.5-Line Pairs per Millimeter (lp/mm) for orthogonal, full-fan CBCT, and half-fan CBCT, respectively. The image registration is accurate to within 1 mm and 1 degree. The data shows consistent imaging-guided system performance with standard deviations in x, y, and z of 0.7, 0.8, and 0.7 mm, respectively. CONCLUSIONS: The system provides excellent image quality and performance, which can be used for IGRT. The proven accuracy of the x-ray imaging and positioning system at McLaren Proton Therapy Center (MPTC) is 1 mm, making it suitable for proton therapy.

Tuning the Magnetic Behavior of Zinc Ferrite via Cobalt Substitution: A Structural Analysis.

Cobalt-doped zinc ferrite is a contemporary material with significant structural and magnetic characteristics. Our study explores the magnetic properties of cobalt-substituted zinc ferrite (ZnxCo1-xFe2O4), synthesized via a simple sol-gel method. By varying the cobalt ratio from 0 to 0.5, we found that zinc substitution impacts both the magnetization and lattice parameters. FTIR analysis suggested the presence of functional groups, particularly depicting an M-O stretching band, within octahedral and tetrahedral clusters. X-ray diffraction analysis confirmed the phase purity and cubic structure. The synthesized materials exhibited an average particle size of 24-75 nm. Scanning electron microscopy revealed the morphological properties, confirming the formation of truncated octahedral particles. In order to determine the stability, mass loss (%), and thermal behavior, a thermal analysis (thermogravimetric analysis (TGA)/differential thermal analysis (DTA)) was performed. The magnetic properties of the synthesized ferrites were confirmed via a vibrating sample magnetometer (VSM). Finally, the highest saturated magnetization and lowest coercivity values were observed with higher concentrations of the cobalt dopant substituting zinc. The synthesized nanomaterials have good stability as compared to other such materials and can be used for magnetization in the near future.

Insights Into Cystic Fibrosis Gene Mutation Frequency, Clinical Findings, and Complications Among Pakistani Patients.

Background Cystic fibrosis (CF) is a genetic disorder with diverse symptoms. Understanding its genetic basis and prevalence is crucial for effective management and treatment. Objective The study aimed to provide comprehensive insights into the frequency of CF gene mutations, clinical presentations, and complications among the Pakistani population. Methodology A cohort comprising 892 patients, ranging in age from 18 to more than 40 years, was selected on the basis of clinical and genetic criteria for the diagnosis of CF. Polymerase chain reaction (PCR) was used to look for 34 variants in the CFTR gene in blood samples. Statistical analysis, which included figuring out the number of mutations, the average age of diagnosis, and the genetic diversity of the samples, was performed to analyze the percentage of patients with specific mutations, offering insights into the genetic diversity. Results In our comprehensive analysis of 892 patient samples, 77.47% (n=691) displayed consanguinity, indicating a family history. The prevailing symptoms included chronic cough (88.67%; n=791), recurrent respiratory infections (76.68%; n=684), and fatigue (73.76%; n=658). The major complications comprised pulmonary infections (22%; n=197), cystic fibrosis-related diabetes (21%; n=187), and malabsorption (20%: n=178). A paired t-test revealed a mean difference of 5.750 with a standard deviation of 9.147, a 95% confidence interval from -0.061 to 11.561, a t-value of 2.178 with 11 degrees of freedom, and a two-tailed p-value of 0.052, suggesting a potential trend towards significance. Nevertheless, the asymptotic significance values of 1.000 and 0.998 for both groups indicate no significant difference. Furthermore, the study identified 12 cystic fibrosis gene mutations, with F508del and N1303K being the most prevalent. Conclusion This research revealed significant consanguinity, confirmed typical CF symptoms, and identified common complications and prevalent CFTR gene mutations (with F508del and N1303K being the most common), providing insights for genetic guidance and treatment in the Pakistani community.

Biocontrol rhizobacteria enhances growth and yield of wheat (Triticum aestivum) under field conditions against Fusarium oxysporum.

The current study aimed to identify the survival of bio-control bacteria with antifungal activity against Fusarium oxysporum and assess their growth promoting activity in wheat crop field conditions. To evaluate the fungicidal activities of isolated bacteria using the dual culture method, both qualitative and quantitative bioassays were performed. Plant Growth Promoting activities such as Indole 3-Acetic Acid (IAA), phosphate solubilization, Hydrogen cyanide (HCN), and Siderophore production were assessed for three biocontrol bacterial isolates (BCB 07, BCB16, and BCB 83) out of 180 with 70% antagonistic activity against Fusarium oxysporum. Chitinase, protease, and cellulase interaction in isolates was also tested. BCB16 was selected as it had 70% antagonist activity against F. oxysporum but also had the highest PGPR (Plant Growth Promoting Rhizobacteria) traits when compared to the other two isolates. BCB16 was also tested for survival in talc powder and in wheat crop field conditions. Even after 4 months in talc powder, the survival rate remained stable. In a wheat crop field, BCB16 reduced the disease incidence of Fusarium oxysporum by 54.38%. When compared to fungus alone treatment, BCB16 increased average yield by 57% alone and 32% in challenged conditions. BCB16 was identified molecularly using the 16s rRNA gene. Bacillus amyloliquefaciens shared 97% of the deduced sequence. The sequence was submitted to genbank and assigned the accession number OM333889. Bacillus amyloliquefaciens has the potential to be used in the field as an alternative to synthetic fungicides against Fusarium oxysporum.

Isolation and characterization of biocontrol bacteria.Molecular Identification of efficient biocontrol bacteria.Survival of biocontrol bacteria in talc powder as carrier material.The Bacillus amyloliquefaciens has plant growth-promoting characteristics.B. amyloliquefaciens reduced the disease incidence of F. oxysporum by 57% in field trials.

Molecular Assessment and Validation of the Selected Enterococcal Strains as Probiotics.

Probiotics are live microorganisms which confer health benefits to the host. Lactic acid bacteria (LAB) are used as probiotics since decades. Enterococci being the member of LAB have proven probiotic strains; therefore, this study was aimed at finding out the potential probiotic candidates from the pool of locally isolated strains. For initial screening, one hundred and twenty-two strains were selected and subjected to different confirmatory and phenotypic tests to choose the best strains that have potential probiotic criteria, i.e., no potential virulence traits, antibiotic resistance, and having tolerance properties. Keeping this criterion, only eleven strains (n = 11) were selected for further assessment. All virulence traits such as production of hemolysin, gelatinase, biofilm, and DNase were performed and not found in the tested strains. The molecular assessment indicates the presence of few virulence-associated genes in Enterococcus faecalis strains with variable frequency. The phenotypic and genotypic assessments of antibiotic resistance profile indicate that the selected strain was susceptible to ten commonly used antibiotics, and there were no transferrable antibiotic resistance genes. The presence of CRISPR-Cas genes also confirmed the absence of antibiotic resistance genes. Various enterocin-producing genes like EntP, EntB, EntA, and EntQ were also identified in the selected strains which make them promising probiotic lead strains. Different tolerance assays like acid, NaCl, and gastric juice tolerance that mimic host conditions was also evaluated by providing artificial conditions. Cellular adhesion and aggregation properties like auto- and co-aggregation were also checked and their results reflect all in the favor of lead probiotic strains.

Circular Production, Designing, and Mechanical Testing of Polypropylene-Based Reinforced Composite Materials: Statistical Analysis for Potential Automotive and Nuclear Applications.

The circularity of polymer waste is an emerging field of research in Europe. In the present research, the thermal, surface, mechanical, and tribological properties of polypropylene (PP)-based composite produced by injection molding were studied. The pure PP matrix was reinforced with 10, 30, and 40% wt. of pure cotton, synthetic polyester, and polyethylene terephthalate post-consumer fibers using a combination of direct extrusion and injection molding techniques. Results indicate that PP-PCPESF-10% wt. exhibits the highest value of tensile strength (29 MPa). However, the values of tensile and flexural strain were lowered with an increase in fiber content due to the presence of micro-defects. Similarly, the values of modulus of elasticity, flexural modulus, flexural strength, and impact energy were enhanced due to an increase in the amount of fiber. The PP-PCCF-40% wt. shows the highest values of flexural constant (2780 MPa) and strength (57 MPa). Additionally, the increase in fiber loadings is directly proportional to the creation of micro-defects, surface roughness, abrasive wear, coefficient of friction, and erosive wear. The lowest average absolute arithmetic surface roughness value (Ra) of PP and PP-PCCF, 10% wt., were 0.19 µm and 0.28 µm. The lowest abrasive wear value of 3.09 × 10-6 mm3/Nm was found for pure PP. The erosive wear value (35 mm3/kg) of PP-PCCF 40% wt. composite material was 2 to 17 times higher than all other composite materials. Finally, the single-step analysis of variance predicts reasonable results in terms of the p-values of each composite material for commercial applications.

Complex spherical fuzzy Aczel Alsina aggregation operators and their application in assessment of electric cars.

The multi-criteria decision-making (MCDM) tool is a robust decision-making technique utilized in several fields like networking, risk management, digital analysis, cybercrime investigation, artificial intelligence, waste management enterprises and many other selection criteria. Complex SFS (CSFS) is a new edition of the spherical fuzzy set (SFS) that offers substantial information about any item in terms of amplitude and phase terms in a wider range of real terms. Complex SFS (CSFS) can be an extension of the spherical fuzzy set (SFS). The Aczel-Alsina aggregation tools are more appropriate aggregation operators (AOs), and they are used to conquer the impact of inconsistent and uncertain data. In this paper, we reveal some new approaches based on Aczel-Alsina aggregation tools under consideration of Complex Spherical Fuzzy (CSF) information. These new approaches include the CSF Aczel-Alsina weighted average (CSFAWA) operator, and the CSF Aczel-Alsina ordered weighted average (CSFOWA) operator. In addition to this, we also introduce a list of novel techniques by making use of the theory of Aczel-Alsina aggregation tools such as CSF Aczel-Alsina weighted geometric (CSFAWG) and CSF Aczel-Alsina ordered weighted geometric (CSFOWG) operators. To demonstrate the resilience and efficacy of the approaches that have been mentioned, we will examine a few exceptional examples and remarkable properties of the methodology that we have devised. In addition, a characterization is provided for an approach to the MCDM issue using the CPF information system. We use the example of electric automobiles as a case study to illustrate the uniformity and dependability of the methodology that we have established. This example was chosen because of the high cost of fuel and the present economic challenges that are being encountered by families in the middle class. An empirical case study is also constructed to determine an electric car that is desirable based on the techniques that we have proposed. To evaluate the correctness and superiority of the established strategies, we compare the outcomes of previously used techniques with the AOs currently being provided.

Synthesis, characterization, and applications of iron oxide nanoparticles.

Objective: The green synthesis method for nanoparticles is getting more attention globally, due to its lesser cost, non-hazardous, and eco-friendly nature. The novelty of the present work is to investigate the anti-bacterial and degradation activity of the green synthesized Iron Oxide NPs. Methods: In this study, the Iron Oxide NPs were synthesized through a green synthesis route from leaves of Ficus Palmata. UV-Vis confirmed Iron Oxide NP's peaks between (230-290 nm), while Fourier transforms infrared spectroscopy analysis showed that several groups were involved in reduction and stabilization. Results: Results indicated that the highest photo thermal activity was shown in light and it was almost 4 folds greater than the control. Similarly, Iron Oxide NPs showed excellent antimicrobial potential against bacterial species "Salmonella typhi" "Xanthomonas Oryzae" and "Lactobacillus" at low concentrations (150 µg/mL). Hemolytic assay results showed that the toxicity was lesser than 5% at both dark and light conditions. Moreover, we also evaluated the photo-catalytic potential of Iron Oxide NPs against methylene orange. Results indicated that almost complete degradation was noted after 90 min in the presence of continuous light. All tests were performed in triplicates. All the data was subjected to P-test (P < 0.5) using Excel and graph pad (V.5.0). Conclusion: Iron Oxide NPs holds a promising future and could be used in treating diseases, and microbial pathogenesis and also could be used as a vector in drug delivery. Moreover, they can also eradicate persistent dyes and could be used as an alternative to remediate pollutants from the environment.

Identification of traits underpinning good breadmaking performance of wheat grown with reduced nitrogen fertilisation.

BACKGROUND: Nitrogen fertiliser is the major input and cost for wheat production, being required to support the development of the canopy to maximise yield and for the synthesis of the gluten proteins that are necessary for breadmaking. Consequently, current high-yielding cultivars require the use of nitrogen fertilisation levels above the yield optimum to achieve the grain protein content needed for breadmaking. This study aimed to reduce this requirement by identifying traits that allow the use of lower levels of nitrogen fertiliser to produce wheat for breadmaking. RESULTS: A range of commercial wheat genotypes (cultivars) were grown in multiple field trials (six sites over 3 years) in the UK with optimal (200 kg Ha-1 ) and suboptimal (150 kg Ha-1 ) application of nitrogen. Bulked grain samples from four sites per year were milled and white flours were baked using three types of breadmaking process. This identified five cultivars that consistently exhibited good breadmaking quality when grown with the lower nitrogen application. Chemical and biochemical analyses showed that the five cultivars were characterised by exhibiting grain protein deviation (GPD) and high dough elasticity. CONCLUSIONS: It is possible to develop novel types of wheat that exhibit good breadmaking quality by selecting for GPD and high dough strength. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Molecular Genetic Analysis of the Autosomal Recessive Non-Syndromic Inherited 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 crumbs homolog 1 (CRB1) and microsatellite marker D8S2332 for gene Retinitis Pigmentosa 1 (RP1). 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 CRB1 gene revealed a homozygous substitution of glycine for valine (c.1152T>G; p.V243G), whereas the RP1 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.

Expression of Chitinase and shRNA Gene Exhibits Resistance to Fungi and Virus.

With the increasing global population, saving crops from diseases caused by different kinds of bacteria, fungi, viruses, and nematodes is essential. Potato is affected by various diseases, destroying many crops in the field and storage. In this study, we developed potato lines resistant to fungi and viruses, Potato Virus X (PVX) and Potato Virus Y (PVY), by inoculating chitinase for fungi and shRNA designed against the mRNA of the coat protein of PVX and PVY, respectively. The construct was developed using the pCAMBIA2301 vector and transformed into AGB-R (red skin) potato cultivar using Agrobacterium tumefaciens. The crude protein extract of the transgenic potato plant inhibited the growth of Fusarium oxysporum from ~13 to 63%. The detached leaf assay of the transgenic line (SP-21) showed decreased necrotic spots compared to the non-transgenic control when challenged with Fusarium oxysporum. The transgenic line, SP-21, showed maximum knockdown when challenged with PVX and PVY, i.e., 89 and 86%, while transgenic line SP-148 showed 68 and 70% knockdown in the PVX- and PVY-challenged conditions, respectively. It is concluded from this study that the developed transgenic potato cultivar AGB-R showed resistance against fungi and viruses (PVX and PVY).

Nutritional probing and storage stability of papaya jam supplemented with date pit powder.

Jam Quality is a factor robustly influenced by storage conditions. The current research aimed to develop papaya jam with improved nutritional attributes, rheological profile, and shelf-life utilizing date pit powder as a functional ingredient. The effect of date pit powder on the formulated product's physicochemical, microbial, and organoleptic properties was analyzed. Results revealed that overall mineral profile (0.35-1.11%), crude fiber (0.56-2.01%), pH (3.51-3.70%), and antioxidant properties (22.97-30.67%) were significantly increased while water activity reduced (0.77-0.73). Moreover, date pit powder improved the color scores like a*(10.10-10.67), b* (8.13-8.78), L* (25.56-28.09), and textural attributes (Cohesiveness: 0.83-0.90; Firmness: 6.82-6.93) of functional papaya jam. Microbial count reduced from 3.60 × 105-3.06 × 105 cfu/ml by adding date pit powder and staying within the acceptable limit (4.13 × 105-3.60 × 105 cfu/ml) during 2-month storage at refrigeration. Organoleptic evaluation depicted that samples treated with date pit powder scored better than the control, and a sample with 75% pectin replacement was considered best.

A Novel Variant in VPS13B Underlying Cohen Syndrome.

Pathogenic variants in vacuolar protein sorting 13 homolog B (VPS13B) cause Cohen syndrome (CS), a clinically diverse neurodevelopmental disorder. We used whole exome and Sanger sequencing to identify disease-causing variants in a Pakistani family with intellectual disability, microcephaly, facial dysmorphism, neutropenia, truncal obesity, speech delay, motor delay, and insomnia. We identified a novel homozygous nonsense variant c.8841G > A: p.(W2947∗) in VPS13B (NM_017890.5) which segregated with the disease. Sleep disturbances are commonly seen in neurodevelopmental disorders and can exacerbate medical issues if left untreated. We demonstrate that individuals with Cohen syndrome may also be affected by sleep disturbances. In conclusion, we expand the genetic and phenotypic features of Cohen syndrome in the Pakistani population.

First molecular confirmation of multiple zoonotic vector-borne diseases in pet dogs and cats of Hong Kong SAR.

In recent years, the incidence of vector-borne diseases (VBDs) has increased throughout the globe. In particular, tick-borne diseases (e.g., caused by Ehrlichia canis, E. ewingii, Anaplasma phagocytophilum, A. platys, Borrelia burgdorferi sensu stricto (s.s.) and Babesia gibsoni) and mosquito-borne diseases (e.g., caused by Dirofilaria immitis) diseases pose a burden on animal health. Nevertheless, there have been no studies undertaken on the occurrence of VBDs in pet dogs and cats in Hong Kong SAR. This study fills this gap, and is the first to determine the seroprevalence of major VBDs, such as those caused by D. immitis, E. canis, E. ewingii, A. phagocytophilum, A. platys and B. burgdorferi s.s, in dogs and cats through commercially available SNAP 4Dx plus testing. Infection by all these pathogens and Babesia sp. was further assessed through PCR and DNA sequencing. A total of 224 blood samples were collected from domestic dogs (n = 159) and cats (n = 65) in Hong Kong SAR during summer 2022. Hematocrit and platelet counts were determined in each blood sample and other hematological parameters were assessed using an automatic hematology analyzer and vortex the specimen for one to two minutes at or near the highest setting to minimize the clumping. All cat sera samples were negative for tested pathogens, but antibodies against some of the pathogens were detected in dog sera samples. Here, the highest figures were recorded for seroprevalence of E. canis/E. ewingii (10.7%), followed by D. immitis (5.7%), and A. phagocytophilum/A. platys (2.5%). No B. burgdorferi s.s. antibodies were detected in any of the dogs tested. Through molecular diagnostics, we detected the presence of B. gibsoni (3.7%), E. canis (3.1%), D. immitis (5.7%), and A. phagocytophilum (1.3%). Neighbor-Joining phylogenetic trees for vector-borne pathogens (i.e., genus Anaplasma sp.) showed 100% clustering to Japan, the USA and Germany, whereas genus Ehrlichia sp. showed 100% clustering to China, Turkey, Cuba, and Greece. Similarly, genus Babesia sp. clustered 100% to India, Sri Lanka and Austria, while D. immitis clustered in Iraq, South Korea, Portugal, France, the USA and Italy. This study provides the first evidence on the occurrence of tick-borne pathogens in pet dogs in Hong Kong SAR. Based on these findings, it is recommended that appropriate screening should be undertaken in domestic dogs to evaluate the prevalence of these pathogens and promote the timely control of VBDs.

In-silico studies of inhibitory compounds against protease enzymes of SARS-CoV-2.

In December 2019, a COVID-19 outbreak caused by SARS-CoV-2 raised worldwide health concerns. In this case, molecular docking and drug repurposing computational approaches were engaged to check the efficiency of plant-based inhibitory compounds against SARS-CoV-2 main protease enzyme and papain-like protease enzyme. Twenty phytochemical inhibitory compounds were collected. Then these compounds were screened based on Lipinski's rule. As a result of this screening eleven compounds were further selected. Quantitative structure-activity relationships analysis was done before molecular docking to check especially the antiviral activity of inhibitory compounds. Docking validation of these compounds was checked by using online server Database of Useful Decoys: Enhanced. Binding affinity value, and pharmacokinetic properties of Aloin compound indicated that it can be used against main protease enzyme of SARS-CoV-2. So, it makes it a promising compound to follow further in cell and biochemical-based assays to explore its potential use against COVID-19.