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Abstract Background Bovine mastitis results in significant economic losses for the dairy industry globally due to milk production losses and decreased herd efficiency. This research aimed to isolate, select, and characterize indigenous lactobacilli with probiotic properties. A total of 40 lactobacilli were isolated from healthy milk samples of cattle and identified at the species level through 16S rDNA sequencing. All isolates were initially screened for antimicrobial activity, and selected isolates underwent in vitro assessment of probiotic properties. Results Among the lactobacilli isolates, varying levels of activity (9 to 19 mm) against cattle mastitogens; Stapylococcus aureus (Staph. aureus), Escherichia coli (E. coli) and Streptococcus dysgalactiae (Strep. dysgalactiae) were observed in the well diffusion assay. These isolates demonstrated auto-aggregation (ranging from 14.29 ± 0.96% to 62.11 ± 1.09%) and co-aggregate (ranging from 9.21 ± 0.14% to 55.74 ± 0.74%) with mastitogens after 2 h. Lactobacillus (Lb.) plantarum CM49 showed sensitivity to most antibiotics tested and exhibited strong inhibitory effects, with mean log10 reductions of 3.46 for Staph. aureus, 2.82 for E. coli, and 1.45 for Strep. dysgalactiae in co-culture experiments. Furthermore, Lb. plantarum CM49 significantly decreased the adhesion rate of mastitogens on the bovine mammary cell line and mouse model, demonstrating its potential effectiveness in preventing mastitis. Conclusion It is concluded that Lb. plantarum CM49 has remarkable probiotic potential with activity against cattle mastitogens in the laboratory and cell culture and competitively excludes mastitogens from bovine mammary cells and ameliorates Staph. aureus-induced mastitis in mice.

Ebola virus disease (EVD) is a rare, highly contagious and a deadly disease with a variable fatality rate ranging from 30% to 90%. Over the past two decades, Ebola pandemic has severely affected the sub-Sahara region including Democratic Republic of the Congo (DRC), and Uganda. The causative agents of the most EVD cases are three distinct species out of six Ebolaviruses namely Zaire Ebolavirus (ZEBOV), Sudan Ebolavirus (SUDV) and Bundibugyo Ebolavirus (BDBV). In recent years, significant strides have been made in therapeutic interventions. Notably, the US Food and Drug Administration has approved two monoclonal antibodies: InmazebTM (REGN-EB3) and Ansuvimab or Ebanga™. Additionally, many small molecules are currently in the developmental stage, promising further progress in medical treatment. Addressing the critical need for preventive measures, this review provides an in-depth analysis of the licensed Ebola vaccines-Ervebo and the combination of Zabdeno (Ad26.ZEBOV) and Mvabea (MVA-BN-Filo) as well as the vaccines which are currently being tested for their efficacy and safety in clinical studies. These vaccines might play an important role in curbing the spread and mitigating the impact of this lethal disease. The current treatment landscape for EVD encompasses both nutritional (supportive) and drug therapies. The review comprehensively details the origin, pathogenesis, and epidemiology of EVD, shedding light on the ongoing efforts to combat this devastating disease. It explores small molecules in various stages of the development, discusses patents filed or granted, and delves into the clinical and supportive therapies that form the cornerstone of EVD management. This review aims to provide the recent developments made in the design and synthesis of small molecules for scientific community to facilitate a deeper understanding of the disease and fostering the development of effective strategies for prevention, treatment, and control of EVD.

Introduction: Undernutrition and tuberculosis (TB) are interrelated, with a bidirectional relationship. Despite its potential impact on nutritional status and TB patient outcomes, there is limited information available on the dietary patterns of TB patients in the Indian TB program. This study aims to address this knowledge gap by evaluating dietary patterns and factors associated with reduced dietary intake among TB patients in a tertiary care center. Methodology: This was a cross-sectional study conducted in a directly observed treatment short center working under the National Tuberculosis Elimination Program. A semi-structured, pretested interviewer-administered questionnaire was used to collect data related to the demographic profile and dietary patterns of 49 patients. Results: The mean calorie intake among the patients was 1525.2 (±897) kcal, whereas the required mean calorie intake was 2231.6 (±409.2) kcal. The mean protein requirement was 66.9 (±15.2) g, whereas the mean protein intake was only 39.2 (±15.4) g. Conclusion: To strive toward the goal of TB elimination in India, the nutritional care of the patient cannot be undermined. The need of the hour is to enforce the nutritional supplementation as well as support program by the government at the national level.

Abstract Thermal evaluation of twin wall panel systems was assessed under vibrant hot and arid conditions of weather in the Arabian Peninsula. Two systems of wall panels (0.6 m × 0.6 m) were prepared. The first system was prepared with a 5.0 cm thick extruded polystyrene (XPS) board. While the second system was prepared with 5.0 cm thick layer of foam-mortar encompassing expanded polystyrene (EPS) beads. Both the thermal insulative layers were sandwiched between two 7.5 cm thick concrete layers. The two wall panel systems were thermally evaluated at the same time in a carefully designed test room. Comparison was accomplished between the two wall systems by measuring the U-value (thermal transmittance) and R-value (resistance). The U-value (air to air) for sandwiched XPS concrete wall system was 0.837 W/m2 K while it was 2.527 W/m2 K for sandwiched EPS beads foam-mortar concrete wall system. The mean U-values (surface to surface) of the sandwiched XPS concrete wall system was 1.143 m2 K/W and 0.293 m2 K/W for sandwiched EPS beads foam-mortar concrete wall system. The sandwiched XPS concrete wall system was more efficient than the sandwiched EPS beads foam-mortar concrete wall system in terms of thermal performance. About 4.5 h of time lag was observed for both the wall panel systems between the external surface temperature and the heat transmission in the internal surface. The output of the FEM simulation by ABAQUS is compared with the measured data for Set-1 (period 16-Aug-2022 to 26-Aug-2022). The hourly temperature change on the outer and inner surfaces has good agreement for both sandwiched XPS concrete wall system and sandwiched EPS beads foam-mortar concrete wall system. The simulation can also predict the heat flux through the two wall systems investigated.

Background: Common mental disorders (CMDs) among adolescents may hamper their psycho-social development. Aim: This study evaluated the prevalence and determinants of CMD like depression and anxiety among late adolescents of an age of 15-–19 years residing in an urban resettlement colony of southeast Delhi. Methods: A community-based cross-sectional study was carried out among 556 randomly selected adolescents. CMD was assessed using Mini International Neuropsychiatry Interview – Kid version 6 (MINI-Kid) based on DSM-IV TR and compliant with ICD-10 definitions of CMD. The associated risk factors were studied using a self-developed semi-structured interview schedule and analyzed using multi-variable logistic regression. Results: A total of 491 adolescents were interviewed (a response rate of 88.3%), of whom 247 (50.3%) were female and 210 (42.8%) belonged to a lower-middle socio-economic status. The lifetime prevalence of CMD was 34% [95% confidence interval (CI): 29.8–38.2]. Of the total, 22.4% (95% CI: 18.7–6.1) of the participants reported depression and 6.7% (95% CI: 4.5–8.9) reported generalized anxiety disorder during their lifetime. Female sex [adjusted odds ratio (aOR) 2.1, 95% CI: 1.4–2.2], experiencing a stressful event in the past 6 months (aOR 4.7, 95% CI: 3.1–7.3), and smoking tobacco (aOR 2.0, 95% CI: 1.2–7.4) significantly increased the odds of having CMD in multi-variate analysis. Conclusion: There is a high prevalence of CMD among adolescents residing in urban resettlement colonies of Delhi, which is composed mostly of people belonging to lower socio-economic strata. Hence, tailored intervention at stress management with promotion of healthy lifestyle is needed for this age group.

Internet of Things (IoT) has become an interesting research domain as numerous devices, preferably equipped with sensors, communication, and actuator modules, are deployed to capture real-time data in the different application areas, such as smart healthcare and industries. These devices have the built-in capacity to directly interact with the physical phenomenon and report any unusual situation within their respective coverage areas, i.e., monitoring a critical patient in the smart hospital but direct communication with a common destination module is not guaranteed and could possibly be very challenging if two or more devices, preferably those in closed proximity, are interested to transmit simultaneously. Therefore, in this manuscript, we are going to present a hybrid slot allocation approach, which is specifically designed for those devices resided in neighborhood and are eager to communication concurrently with a common destination device, i.e., server. In the beginning, the k-mean clustering algorithm is used to group these devices into clusters where server is forced to collect data from devices deployed in the respective coverage areas. Thus, every server generates dedicated slots for active devices and an additional slot for server(s). Similarly, the proposed neighborhood-enabled time division multiple access (TDMA) has the flexibility of assigning multiple slots to a requesting device if available, which is needed in scenarios, such as detection of pest in the field. Additionally, a member device is allowed to migrate (if needed and possible) from one server's coverage region to another. Simulation results confirmed that the proposed approach is better than the existing algorithms (opportunistic TDMA, hybrid TDMA, and non-orthogonal multiple access), particularly in terms of bandwidth, end-to-end delay, and empty slot utilization. The proposed scheme has improved bandwidth and empty slot utilization, which are approximately 15% and 12%, respectively, whereas it has achieved approximately 94.89% utilization of the available slots which was previously 93.4%.

Abstract Background Randomized controlled trials and real-world studies suggest that combination therapy with sodium–glucose transport protein 2 inhibitors (SGLT2is) and glucagon-like peptide-1 receptor agonists (GLP-1RAs) is associated with improvement in fasting plasma glucose (FPG), glycated hemoglobin (HbA1c), systolic blood pressure (SBP), body mass index (BMI), and total cholesterol levels. However, a systematic review of available real-world evidence may facilitate clinical decision-making in the real-world scenario. This meta-analysis assessed the safety and effectiveness of combinations of SGLT2is + GLP-1RAs with a focus on their cardioprotective effects along with glucose-lowering ability in patients with type 2 diabetes mellitus (T2DM) in a real-world setting. Methods Electronic searches were performed in the PubMed/MEDLINE, PROQuest, Scopus, CINAHL, and Google Scholar databases. Qualitative analyses and meta-analyses were performed using the Joanna Briggs Institute SUMARI software package and Review Manager v5.4, respectively. Results The initial database search yielded 1445 articles; of these, 13 were included in this study. The analyses indicated that SGLT2is + GLP-1RAs combinations were associated with significantly lower all-cause mortality when compared with individual therapies (odds ratio [95% confidence interval [CI] 0.49 [0.41, 0.60]; p

Abstract Objective Multi-drug resistance (MDR) has notably increased in community acquired uropathogens causing urinary tract infections (UTIs), predominantly Escherichia coli. Uropathogenic E. coli causes 80% of uncomplicated community acquired UTIs, particularly in pre-menopausal women. Considering this high prevalence and the potential to spread antimicrobial resistant genes, the current study was conducted to investigate the presence of clinically important strains of E. coli in Pakistani women having uncomplicated cystitis and pyelonephritis. Women belonging to low-income groups were exclusively included in the study. Seventy-four isolates from urine samples were processed, phylotyped, and screened for the presence of two Single Nucleotide Polymorphisms (SNPs) particularly associated with a clinically important clonal group A of E. coli (CgA) followed by antibiotic susceptibility testing and genome sequence analysis. Results Phylogroup B2 was most prevalent in patients and 44% of isolates were positive for the presence of CgA specific SNPs in Fumarate hydratase and DNA gyrase subunit B genes. Antibiotic susceptibility testing showed widespread resistance to trimethoprim-sulfamethoxazole and extended-spectrum beta-lactamase production. The infection analysis revealed the phylogroup B2 to be more pathogenic as compared to the other groups. The genome sequence of E. coli strain U17 revealed genes encoding virulence, multidrug resistance, and host colonization mechanisms. Conclusions Our research findings not only validate the significant occurrence of multidrug-resistant clonal group A E. coli (CgA) in premenopausal Pakistani women suffering from cystitis and pyelonephritis but also reveal the presence of genes associated withvirulence, and drug efflux pumps. The detection of highly pathogenic, antimicrobial-resistant phylogroup B2 and CgA E. coli strains is likely to help in understanding the epidemiology of the pathogen and may ultimately help to reduce the impact of these strains on human health. Furthermore, the findings of this study will particularly help to reduce the prevalence of uncomplicated UTIs and the cost associated with their treatment in women belonging to low-income groups.

Abstract The Rhus gall aphid, Schlechtendalia chinensis, feeds on its primary host plant Rhus chinensis to induce galls, which have economic importance in medicines and the food industry. Rhus gall aphids have a unique life cycle and are economically beneficial but there is huge gap in genomic information about this group of aphids. Schlechtendalia chinensis induces rich-tannin galls on its host plant and is emerging as a model organism for both commercial applications and applied research in the context of gall production by insects. Here, we generated a high-quality chromosome-level assembly for the S. chinensis genome, enabling the comparison between S. chinensis and non-galling aphids. The final genome assembly is 344.59 Mb with 91.71% of the assembled sequences anchored into 13 chromosomes. We predicted 15,013 genes, of which 14,582 (97.13%) coding genes were annotated, and 99% of the predicted genes were anchored to the 13 chromosomes. This assembly reveals the endogenization of parvovirus-related DNA sequences (PRDs) in the S. chinensis genome, which could play a role in environmental adaptations. We demonstrated the characterization and classification of cytochrome P450s in the genome assembly, which are functionally crucial for sap-feeding insects and have roles in detoxification and insecticide resistance. This genome assembly also revealed the whole genome duplication events in S. chinensis, which can be considered in comparative evolutionary analysis. Our work represents a reference genome for gall-forming aphids that could be used for comparative genomic studies between galling and non-galling aphids and provides the first insight into the endogenization of PRDs in the genome of galling aphids. It also provides novel genetic information for future research on gall-formation and insect-plant interactions.

A D.C. motor's rotational speed is regulated in this study using a PID controller and a fuzzy logic controller. In contrast to the fuzzy logic controller, which uses rules based on knowledge and experience, the proportional-integral-derivative (PID) controller requires a mathematical system model. This study investigates the regulation of a DC motor's velocity using PID and fuzzy logic controllers. The PID controller utilizes a mathematical model and parameter tuning by trial and error. Still, the fuzzy logic controller (FLC) operates on rule-based knowledge, enabling it to handle the nonlinear features of the DC motor effectively. The FLC design entails intricate determinations, including the establishment of a rule base and the process of fuzzification. A total of 49 fuzzy rules have been devised to achieve precise control. Based on MATLAB/SIMULINK simulations, the study concludes that the Fuzzy Logic Controller (FLC) beats the Proportional-Integral-Derivative (PID) controller. The FLC exhibits superior transient and steady-state responses, shorter response times, reduced steady-state errors, and higher precision. This study emphasizes the efficacy of the FLC (Fuzzy Logic Controller) in dealing with the difficulties associated with DC motor control. It presents a strong argument for the suitability and efficiency of FLCs in industrial environments compared to conventional PID (Proportional-Integral-Derivative) controllers. There are a wide variety of ways to construct a fuzzy logic controller. The speed error and the rate of change in the speed error are two inputs to the FLC. Defuzzification is done by focusing on the core of the problem. The results show that FLC is superior to PID controllers in efficiency and effectiveness due to its reduced transient and steady-state factors.

Objective: To assess the relationship between the fingerprint patterns and academic performance of students in Anatomy. Methodology: This Cross-sectional study was conducted in the Anatomy department of Islam Medical and Dental College Sialkot. The duration of the study was from mid-November 2022 to mid-December 2022. A total of 300 healthy medical (1st and 2nd year MBBS) and dental (1st year BDS) students with no injury and deformity, voluntarily participated in this study. The student’s age were 20-22 years. Simple convenient sampling was used. Ethical clearance was obtained before study. Thumb prints were collected along with questionnaire. The study was done by categorizing the pattern into arches, loops and whorls. The academic performance was taken in percentage and classified as above 70 %, between 50 % and 70 % and below 50%. The statistical analysis of categorical data was done by using non-parametric test i.e. chi square test. A P value of < 0.05 was considered statistically significant. Results: The most common pattern observed among students was loops. The students with loop pattern were in high scoring group while students with whorl and arch pattern scored less marks in Anatomy. Conclusion: The percentage of academic performance was higher in students with loops as compared to whorl and arch.

This research paper reports an enhancement of thermal, optical, mechanical and antibacterial activities of the Polyvinyl alcohol–Nanodiamonds (PVA-NDs) composite required for the food packaging industry. The synthesis of composites was done by the wet processing method. The large surface area of NDs facilitated the robust interaction between the hydroxyl group and macromolecular chains of PVA to enhance the hydrogen bonding of PVA with NDs rather than PVA molecules. Thus, a reduction in PVA diffraction peak intensity was reported. NDs improved the thermal stability by preventing the out-diffusion of volatile decomposition products of PVA. The results also revealed an enhancement in tensile strength (∼60 MPa) and ductility (∼180 %). PVA-NDs composite efficiently blocked the UVC (100 %), most of the part of the UVB (∼85 % above 300 nm), and UVA (∼58 %). Furthermore, enhanced antibacterial activities were reported for PVA-NDs composite against E. coli and S. aureus. NDs accumulated around the bacterial cells prevented essential cellular functions and led to death. Hence, this composite could be a promising candidate for safe, thermally stable, strong, flexible, transparent, UV- resistant antibacterial food packaging material.

This research investigates the impact of sea level rise (SLR) on the Indus Delta, a vital ecosystem increasingly vulnerable to climate change repercussions. The objective of this study is to comprehensively assess the flooded areas under various shared socioeconomic pathway (SSP) scenarios based on the Intergovernmental Panel on Climate Change's (IPCC) 6th Assessment Report. The study employs a GIS-based bathtub model, utilizing historical (1995–2014) and IPCC-projected (2020–2150) tide gauge data from Karachi, Kandla, and Okha stations to identify potential inundated areas threatened by coastal flooding. Additionally, it analyzes LANDSAT-derived multispectral images to identify coastal erosion hotspots and changes in the landscape. A supervised random forest classifier is used to classify major landforms and understand alterations in land cover. Furthermore, neural network-based cellular automata simulations are applied to predict future land cover for 2050, 2100, and 2150 at risk of inundation. The results indicate that under different SSP scenarios, the estimated inundated land area varies from 307.36 km2 (5 % confidence on SSP1-1.9) to 7150.8 km2 (95 % confidence on SSP5-8.5). By 2150, the region will lose over 550 km2 of agricultural land and 535 km2 of mangroves (mean SLR projection). This work emphasizes identifying sensitive land cover for SLR-induced coastal flooding. It might fuel future policy and modeling endeavors to reduce SLR uncertainty and build effective coastal inundation mitigation methods.

Introduction: The Macrophage Migration Inhibitory Factor (MIF), a key pro-inflammatory mediator, is responsible for modulating immune responses. An array of inflammatory and autoimmune diseases has been linked to the dysregulated activity of MIF. The significance in physiological as well as pathophysiological phenomena underscores the potential of MIF as an attractive target with pharmacological relevance. Extensive research in past has uncovered a number of inhibitors, while the ISO-1, or (S, R)-3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid methyl ester being recognized as a benchmark standard so far. Recent work by Yang and coworkers identified five promising dietary flavonoids, with superior activity compared to the standard ISO-1. Nevertheless, the exact atomic-level inhibitory mechanism is still elusive.Methods: To improve the dynamic research, and rigorously characterize, and compare molecular signatures of MIF complexes with ISO-1 and flavonoids, principal component analysis (PCA) was linked with molecular dynamics (MD) simulations and binding free energy calculations.Results: The results suggest that by blocking the tautomerase site these small molecule inhibitors could modify the MIF activity by disrupting the intrinsic dynamics in particular functional areas. The stability matrices revealed the average deviation values ranging from 0.27–0.32 nm while the residue level fluctuations indicated that binding of the selected flavonoids confer enhanced stability relative to the ISO-1. Furthermore, the gyration values extracted from the simulated trajectories were found in the range of 1.80–1.83 nm.Discussion: Although all the tested flavonoids demonstrated remarkable results, the one obtained for the potent inhibitors, particularly Morin and Amentoflavone exhibited a good correlation with biological activity. The PCA results featured relatively less variance and constricted conformational landscape than others. The stable ensembles and reduced variation in turns might be the possible reasons for their outstanding performance documented previously. The results from the present exploration provide a comprehensive understanding of the molecular complexes formed by flavonoids and MIF, shedding light on their potential roles and impacts. Future studies on MIF inhibitors may benefit from the knowledge gathered from this investigation.

Bacterial antimicrobial resistance (BAMR) seems to pose the greatest threat to public health, food safety, and agriculture in this century. The development of novel efficient antimicrobial agents to combat bacterial infections has become a global issue. Silver nanoparticles (Ag NPs) appeared as a feasible alternative to antibiotics. However, Ag NPs face cost, toxicity, and aggregation issues which limit their antibacterial activity. This work aims to stabilize Ag NPs with enhanced antimicrobial activity at comparatively lower Ag concentrations to prevent bacterial infections. For this purpose, the Ag core was covered with nanodiamonds (NDs). Ag-NDs composite have been synthesized by microplasma technique. TEM analysis confirmed the presence of both Ag and NDs in the Ag-NDs composite. A particle size (∼19 nm) was reported for Ag-NDs at the highest concentration as compared to Ag NPs (∼3 nm). The conduction band of the diamond acted as an extremely strong reducing agent for Ag NPs. The large surface area of NDs stabilized the Ag NPs. A redshift (∼400 nm–406 nm) in UV–visible spectra of the Ag-NDs composite indicated the formation of bigger-sized Ag NPs after incorporating NDs. XRD and LIBS analysis verified the increase in intensity of Ag-NPs by increasing ND concentration. The presence of functional groups including OH, CH, and Ag/Ag2O was confirmed by FTIR. Bacterial inhibition growth appeared to be a dose-dependent process. The minimum inhibition concentration value of Ag-NDs composite at the highest NDs concentration against E. coli (∼ 0.69 μg/ml) and S. aureus (∼44 μg/ml). This is the first study to report the smallest MIC for E. coli (

Abstract Blackcurrant possesses various health‐endorsing attributes owing to its polyphenol profile. Recent studies have demonstrated its therapeutic potential against various health disorders. Various bioactives present in blackcurrants have different functional and pharmacological aspects including anti‐inflammatory, antioxidant, and antimicrobial properties. The most dominant and important bioactive include anthocyanins, flavonols, phenolic acids, and polyunsaturated fatty acids. Food formats derived from blackcurrants comprise pomace, juice, powder, and extracts. All these food formats have industrial, prebiotic, and pharmacological benefits. In the current article, the nutritional composition, industrial applications, and therapeutic potential are discussed in the recent literature. Moreover, novel extraction techniques for the extraction of bioactive compounds present in blackcurrants and their safety concerns have been elaborated.

Abstract There is an increasing challenge in probiotic viability and stability during food product formulation, processing, and storage. However, synbiotic functional foods have promising potential to deliver the targeted benefits. This study aimed to isolate the okara from soybean residue, and obtained okara flour was further characterized using Fourier transform infrared spectroscopy (FTIR) and scanning electron microscope (SEM). Synbiotic ice cream was developed by fortification with Lactobacillus rhamnosus GG and okara at different concentrations (1–3%). Additionally, the synbiotic ice cream was subjected to physicochemical and sensory attributes over 60 days of storage. High viability of L. rhamnosus GG (8.17 log CFU/mL) was observed during storage at 3% okara. Moreover, adding okara at 2% or higher improved viscosity, reduced overrun, and maintained probiotic viability. When compared to the control (ice cream without okara), synbiotic ice cream exhibited a higher protein content and a lower fat level. The synergistic combination of probiotics and okara in ice cream is a potentially novel approach for developing functional ice cream. The addition of okara is not only helpful in increasing the nutritional value of the ice cream but will also be a way forward to minimize agricultural waste. Synbiotic ice cream developed in this study may be considered a potential functional food rich in protein and low in fat.

Background and Objectives: Acinetobacter baumannii (A. baumannii), particularly carbapenem-resistant A. baumannii (CRAB), represents a grave concern in healthcare settings and is associated with high mortality. This study aimed to conduct molecular, mutational, and phylogenetic analyses of specific genes in CRAB and evaluate the synergistic effects of selected antimicrobial combinations. Materials and Methods: Phenotypic characterization was performed on six CRAB strains by using the Modified Hodge Test (MHT) and IMP-EDTA Double-Disc Synergy Test (IMP-EDTA DDST). Carbapenemase- and metallo-beta-lactamase-encoding genes were amplified by using Polymerase Chain Reaction. Phylogenetic analysis using the MEGA 11 tool was used to determine the evolutionary relatedness of these genes. Mutational analysis was performed by using I-Mutant, MUPro, and PHD-SNP bioinformatics tools to predict mutations in the carbapenemase-encoding genes. Microdilution checkerboard titration assessed the synergistic effects of antimicrobial combinations (azithromycin–meropenem, rifampicin–meropenem, meropenem–colistin, and azithromycin–colistin) on these CRAB isolates. Results: The phenotypic characterization of six CRAB isolates revealed positive results for MHT and IMP-EDTA DDST. The molecular characterization revealed that carbapenemase- and MBL-encoding genes were present in all isolates with varying frequencies, including blaOXA-51 (100%) and blaIMP (0%). The sequence analysis revealed high evolutionary relatedness to sequences in the NCBI database. The mutational analysis identified 16 mutations, of which 1 mutation (P116L) in the blaOXA-58 gene predicted a change in the protein product, potentially contributing to carbapenem resistance. The checkerboard titration method did not reveal any synergism among the tested antimicrobial combinations against CRAB. Conclusion: This study’s findings underscore the significant challenges posed by CRAB isolates harboring multiple resistant genes in treatment. This highlights the urgent need for novel antimicrobial agents, a crucial step towards reducing mortality rates not only in Pakistan but also globally.

Herbal medicinal plants have been used for centuries in traditional medicine, and it is interesting to see how modern research has identified the active compounds responsible for their therapeutic effects. The green synthesis of silver nanoparticles using herbal medicinal plants, such as Swertia chirata, is particularly noteworthy due to its antimicrobial properties. In the current study, the Swertia chirata plant was collected for the first time from the region of Murree, Punjab, Pakistan. After collection, extracts were prepared in different solvents (ethanol, methanol, chloroform, and distilled water), and silver nanoparticles were synthesized by reducing silver nitrate (AgNO3). The UV–visible spectrophotometer, SEM, and EDX were used to characterize the synthesized nanoparticles in terms of their size and shape. The phytochemical analysis of crude extract was performed to determine the presence of different kinds of phytochemicals. The antibacterial activity of plant extracts and the silver nanoparticles were then assessed using the agar well diffusion method against various pathogenic bacteria. The results showed that the plant contains several phytochemicals with remarkable antioxidant potential. The antibacterial analysis revealed that silver nanoparticles and the plant extracts exhibited a significant zone of inhibition against human pathogenic bacteria (Escherichia coli, S. capitis, B. subtilis, and Pseudomonas aeruginosa) as compared to the cefixime and norfloxacin. This implies that the nanoparticles have the potential to be used in nano-medicine applications, such as drug delivery systems, as well as for their antibacterial, antifungal, and antiviral activities. Additionally, the development and application of materials and technologies at the nanometer scale opens possibilities for the creation of novel drugs and therapies. Overall, the study highlights the promising potential of herbal medicinal plants found in Murree, Punjab, Pakistan, and green-synthesized silver nanoparticles in various fields of medicine and nanotechnology.

The primary objective of this investigation was to assess the viability of free and encapsulated Lactobacillus plantarum probiotics in mango juice and under simulated gastrointestinal conditions. Specifically, the probiotics were encapsulated using sodium alginate and alginate-soy protein isolate through the internal gelation method, and the obtained probiotics were characterized for various attributes. Both free and encapsulated probiotics were exposed to challenging conditions, including thermal stress, low temperature, and simulated gastrointestinal conditions. Additionally, both types of probiotics were incorporated into mango juice, and their survival was monitored over a 28-day storage period. Following viability under simulated gastrointestinal conditions, the count of free and encapsulated probiotic cells decreased from initial levels of 9.57 log CFU/mL, 9.55 log CFU/mL, and 9.53 log CFU/mL, 9.56 log CFU/mL to final levels of 6.14 log CFU/mL, 8.31 log CFU/mL, and 6.24 log CFU/mL, 8.62 log CFU/mL, respectively. Notably, encapsulated probiotics exhibited a decrease of 1.24 log CFU and 0.94 log CFU, while free cells experienced a reduction of 3.43 log CFU and 6.24 log CFU in mango juice over the storage period. Encapsulated probiotics demonstrated higher viability in mango juice compared to free probiotics throughout the 28-day storage period. These findings suggest that mango juice can be enriched with probiotics to create a health-promoting beverage.

ABSTRACTStarch nanoparticles resist digestion in the upper intestine but not in the colon. They are proven to exert prebiotic effects on the human body. Starch-based nanoparticles have generated a great interest in the food and agriculture sector due to their biocompatibility, wide array of natural sources and ease of modification. In the current study, two starch sources (rice and water chestnut) were used to synthesize nanoparticles for the encapsulation of probiotic bacteria. Obtained nanoparticles were characterized for their morphological, molecular, and structural attributes using Fourier Transform Infrared Spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray diffraction (XRD), and zeta sizer. Furthermore, free and nano-encapsulated probiotics were subject to simulated gastrointestinal digestion conditions. The average size of rice and water chestnut starch-based nano-capsules ranged from 309 to 427 nm. The encapsulation efficiency was recorded for rice and water chestnut starch as 89% and 95%, respectively. SEM micrographs exhibited entrapment of probiotics in wall materials. The surface of the capsules showed tiny, smooth surface polygonal granules. XRD images showed loss of crystallinity structure following encapsulation. Higher probiotic viability was recorded under simulated gastrointestinal conditions for nano-encapsulated probiotics compared to free probiotics. Conclusively, water chestnut and rice-based starch nanoparticles showed overall the best results regarding the viability of probiotics under stressed conditions.

ABSTRACTProbiotics possess many health-endorsing properties; however, their viability and stability under detrimental condition is uncertain. Encapsulation technology provides protection under various stressed conditions. Furthermore, combination of different wall materials ensures the target delivery of core materials. In the current study, probiotic was encapsulated using solid lipid micro particles (SLMP) that were prepared by high shear homogenization. Whey protein and gum Arabic were used as wall material to encapsulate the probiotics. Obtained micro beads were subjected for size, zeta potential, and encapsulation efficiency measurement. Scanning electron microscopy was used for morphological characterization of the microbeads. Molecular characterization of obtained micro beads was done by Fourier-transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD). Furthermore, the viability and stability were assessed under simulated gastrointestinal conditions. Free and encapsulated probiotics were incorporated in chocolate to evaluate the stability of probiotics. The results in this study indicated that encapsulated probiotics showed significant (P

ABSTRACTThe present study aimed to evaluate the effect of the incorporation of protein-based nanoencapsulation on the viability and stability of probiotic Lactobacillus rhamnosus in digestion conditions and model food. In the study’s first phase, protein-based nanoparticles were prepared by the pH cycling method, and then the probiotic, Lactobacillus rhamnosus, was nano-encapsulated. Two types of proteins, namely, whey protein and zein protein, were used to encapsulate probiotics individually. The obtained nano-encapsulated probiotics were characterized by performing particle size, SEM, FTIR, and in vitro assay was performed. Then, free and nano-encapsulated probiotics were added to the model food (yogurt) and analyzed for microbiological and sensory evaluation. Nanoencapsulation with both types of proteins significantly (p

ABSTRACTThe demand for probiotic-based functional food is increasing globally owing to its health-endorsing attributes. There are various driving forces behind probiotic therapy. However, Intestinal dysbiosis in humans is the prime driving force behind this increasing trend in the consumption of probiotic-based functional food. Probiotics have numerous health potentials, however, their target delivery and stability is a great challenge for food manufacturer. Microencapsulation with various types of coating materials is trending for the target and stable delivery of potential probiotics. There are various encapsulation techniques with pros and cons. The type of probiotic bacteria, encapsulation methods, and coating materials are considered crucial factors to prolong the viability of probiotics under hostile conditions. The current review addresses the opportunities, challenges, and future trends surrounding matrix materials used in probiotic encapsulation. The review also describes the current studies and their findings on the various types of encapsulation materials. This comprehensive review could be a way forward in the selection of efficient and effective wall material for the target delivery of sensitive ingredients.

Waste foundry sand (WFS), a by-product of the casting industry, is a potential material that may be employed as a substitute for fine aggregate in concrete. In the present study, gene expression programming (GEP) and multi-expression programming (MEP) are used to generate predictive models for the split tensile strength (STS) and elastic modulus (E) of waste foundry sand concrete (WFSC). Therefore, a comprehensive database was collected that contains 146 and 242 values of E and STS, respectively. Seven different variables were chosen as input for the development of the ML-based models. The reliability and accuracy of the proposed model were evaluated by using various statistical indicators. Given the performance assessment, both GEP and MEP accurately predict the E with a correlation of 0.994 and 0.996, respectively. However, GEP performance was much superior in predicting STS (R = 0.987) as compared to the MEP model (R = 0.892). The integrated statistical performance (ρ, OF) of both models approaches zero, indicating the excellent performance and generalization potential of the developed models. For the interpretation of machine learning (ML) models, Shapley additive explanation (SHAP) was used to know about the input variables' importance and influence on the output parameter. The SHAP analysis revealed that a higher ratio of FA/TA results in the enhancement of the elastic modulus, whereas CA/C higher ratio is favorably influencing the split tensile strength up to some extent, however, this trend changes when the ratio is further increased. These soft computing prediction techniques can incentivize the use of WFS in sustainable concrete, reducing waste disposal and promoting environment-friendly construction. Furthermore, it is recommended that the findings of this study be validated with more extensive data sets and that other ML techniques be investigated.

OBJECTIVES To determine the association between time to thrombolysis and left ventricular (LV) systolic function in patients with ST-elevation Myocardial Infarction (STEMI). METHODOLOGY T This descriptive case series study compared the time to thrombolysis and LV systolic function of 149 STEMI patients. LV systolic function was evaluated following thrombolysis, and data were collected and analyzed using SPSS version 21.0 software. This study was conducted at the Ayub Teaching Hospital Cardiology Department in Abbottabad from August 2022 to January 2023. RESULTS The study examined how LV systolic function and symptoms in STEMI patients related to when thrombolysis was administered. The 149 patients analyzed included 28 with normal LV function, 86 with mildly impaired function, 26 with moderately impaired function, and 9 with severely impaired function. In contrast to delayed thrombolysis, which was linked to higher levels of impairment, immediate thrombolysis was correlated with a higher percentage of normal LV function. The prevalence of patients with severely compromised LV function was also higher in those who received thrombolysis within 3 to 6 hours and 9 to 12 hours. LV function was correlated with the type of myocardial infarction, with varying degrees of impairment seen in AWMI and IWMI patients. Gender had no discernible effect on LV function. CONCLUSION Improved LV function is associated with early thrombolysis within hours of STEMI symptoms, highlighting the significance of prompt intervention and minimizing wait times for better patient outcomes.

Introduction: Present study was designed to understand the role of inflammatory markers and growth factors in later stage of wound healing after treatment with SPPE on wound in alloxan induced diabetic rats. Methodology: The study was performed by inducing diabetes in wistar rats using alloxan at 150 mg/kg, subcutaneously. The animals were allocated in three groups: Group 1- diabetic excision wound (DEW) without any treatment, Group 2- DEW treated with gel vehicle only, Group 3- DEW treated with 5 %SPPE containing gel. During the treatment period of twenty-one days animals were monitored based on clinical signs, morbidity, and mortality. Percent wound contraction, Growth factors (VEGF, and EGF), proinflammatory markers TNF- α, IL-1β, MCP-1, IL-4, and IL-17a, anti-inflammatory marker IL-10 in wound lysates, and histology of skin were all considered as indicators of effectiveness. Results: The vehicle control when compared with the SPPE in gel (5.0 g extract per 100 g of gel) presented significant [p

AbstractThe fruits of citrus family are rich in flavonoids with superior antioxidant properties; citrus bioactive components are potentially beneficial without adverse effects or cytotoxicity in healthy individuals. Citrus fruit peel contains ample amount of hesperidin which is flavanone glycoside and has showed effective role in biological systems as an antioxidant and anti-inflammatory bioactive compound. Different studies have investigated such roles of hesperidin. Due to superior antioxidant potential of hesperidin and its derivatives, these compounds have been intensively studied for their anti-cancer and cardio protective perspective. This review article is a collection of updated data covering different aspects of hesperidin and its derivatives, like occurrence, physical and chemical composition, pharmacokinetics and safety of these compounds. Moreover, effective role of these compounds against different lifestyle-related metabolic syndromes in mechanistic perspective has also been extensively discussed in this article.

Objectives: Adenosine receptor signaling and suppressing potential pathways such as the aryl hydrocarbon receptor (AHR) and takeda G-protein-coupled receptor-5(TGR5) have been identified as potential targets for enhancing metabolic health. Certain adenosine receptor (AR) ligands have been suggested to reduce inflammation and improve thermogenesis in adipose tissue. Methods: This study employed in-silico biomolecular fractions of adenosine receptors and other potential targets to understand the mechanism of action of Asperuloside. Additionally, the anti-obesity potential of Asperuloside, a dual-acting ligand with A2A adenosine receptor (A2AAR) agonist and A3 adenosine receptor (A3AR) agonist activities, were examined using computational analysis in the obesity model. The impact of Asperuloside on inflammation and thermogenesis was studied through diverse protein structures such as the A2AAR complex with agonist/A2AAR complex with antagonist, the rhodopsin mutant with bound galphact peptide (as A3 adenosine receptor), The Human TGR5 complex with synthetic agonist 23H, and AHR receptors antagonism. Results: The study found that Asperuloside has therapeutic affinity for the binding site of adenosine receptors and revealed a novel binding interaction that helps reduce inflammation and improve thermogenesis-mediated obesity. Conclusion: Asperuloside may have anti-obesity effects through its dual-acting ligand with A2AAR and A3AR agonist activities. This study provides a major step towards understanding the mechanism of action of Asperuloside and its potential use as an anti-obesity drug. In vivo tests will help ascertain its pharmacokinetic characteristics, metabolite production in animals, and the effects of chronic daily absorption.

ABSTRACT Aspergillus fumigatus is a respiratory pathogen, which has the potential to cause brooder pneumonia in poultry chicks and aspergillosis in birds and farmers. Frequent and irrational use of antifungals has led to the development of resistance in the fungal species. The present study aimed to evaluate the antifungal activity of essential oils (EOs) from four different spices against multi-drug resistant (MDR) A. fumigatus isolated from 40 poultry feed samples. Twenty fungal isolates were identified to be A. fumigatus based on the macroscopic and microscopic characteristics, which were confirmed by targeting and amplifying the species-specific rodA gene (313 bp). Five (5/20) isolates were found to be MDR by the disk diffusion method. EOs were tested as an alternative strategy for 5 MDR isolates of A. fumigatus. Clove EO provided better activity by 6 log10 reduction for 30-90 min exposure as compared to other EOs, therefore, shortlisted for well-diffusion and minimum inhibitory concentration (MIC) method. Clove EO provided promising results as compared to fluconazole, amphotericin B, ketoconazole, nystatin and voriconazole as its mean ZOI (39.6 ± 6.34) were better than all other commercial antifungals (p< 0.001). Mean MIC was reported to be 0.195 ± 0.11 µl/ml. Safety testing of the EO produced 74-82% cell viability for the range of 0.390 - 0.097 µl/ml as compared to DMSO. Findings of the present study highlight that clove EO can be source of a good therapeutic agent for infections of MDR isolates of A. fumigatus.

CRISPR-Cas9, the “genetic scissors”, is being presaged as a revolutionary technology, having tremendous potential to create designer crops by introducing precise and targeted modifications in the genome to achieve global food security in the face of climate change and increasing population. Traditional genetic engineering relies on random and unpredictable insertion of isolated genes or foreign DNA elements into the plant genome. However, CRISPR-Cas based gene editing does not necessarily involve inserting a foreign DNA element into the plant genome from different species but introducing new traits by precisely altering the existing genes. CRISPR edited crops are touching markets, however, the world community is divided over whether these crops should be considered genetically modified (GM) or non-GM. Classification of CRISPR edited crops, especially transgene free crops as traditional GM crops, will significantly affect their future and public acceptance in some regions. Therefore, the future of the CRISPR edited crops is depending upon their regulation as GM or non-GMs, and their public perception. Here we briefly discuss how CRISPR edited crops are different from traditional genetically modified crops. In addition, we discuss different CRISPR reagents and their delivery tools to produce transgene-free CRISPR edited crops. Moreover, we also summarize the regulatory classification of CRISPR modifications and how different countries are regulating CRISPR edited crops. We summarize that the controversy of CRISPR-edited plants as GM or non-GM will continue until a universal, transparent, and scalable regulatory framework for CRISPR-edited plants will be introduced worldwide, with increased public awareness by involving all stakeholders.

With an increase in the progression of research and development in the medical field, the experimental use of animals for the efficacy and safety testing of pharmaceuticals is on rise. Every year, millions of animals are used for experimental testing during which these suffer from pain and are then eventually sacrificed. Besides bioethical issues, animal experimentation is associated with many disadvantages like high cost, the requirement of skilled manpower, approval, and is time-consuming. Therefore, attempts have been made by researchers to design and develop a number of alternative methods that could bypass animal experiments. These methods not only give accurate results but can also save lives of millions of animals annually. Research techniques, including computer and robotics together with molecular biology techniques, are applied to discover new methods to replace animal testing. Several alternative methods are discussed in this review. Some of these methods can predict the behavior of drugs accurately and are as reliable as in-vivo animal models. Furthermore, these alternative methods offer a variety of advantages over experimental animals. However, there is still a great need to discover and develop new, accurate, and reliable methods to replace experimental animals.

Clostridium perfringens toxinotype B is causative agent of lamb dysentery in Pakistan. For assessment of gene expression, it is preferred to optimize toxin production potential of indigenous isolates. Current study aimed to confirm the local isolates of C. perfringens toxinotype B on molecular basis followed by optimization of major toxins under varied physical and chemical conditions. The isolates were identified using microbiological and biochemical assays and confirmed by PCR followed by nucleotide sequencing of 16S rRNA amplified gene. These nucleotide sequences were submitted to NCBI GenBank® and accession numbers retrieved were MW867097, MW867098, MW867099, MW867100 and MW867101. Confirmation of toxinotype B was done by PCR amplification of alpha beta and epsilon toxin genes. Major toxins were optimized at varied physical (temperature and pH) and chemical (reinforced clostridial media, Robertson"s cooked meat media, egg meat media, fluid thioglycolate media and iron milk media) conditions. Higher hemolytic units of alpha toxin (21.45±0.53 HU/mL), epsilon toxin (16.57±0.19 HU/mL) and higher cytotoxic units of beta toxin (18.65±0.34 HU/mL) were produced at 37°C with pre-adjusted pH 6.0 in Robertson cooked meat media. The ELISA percentages of alpha (14.27%, 13.92%, 13.67%, 13.56%, 13.45%), beta (12.43%, 12.81%, 12.39%, 12.61% and 13.07%) and epsilon (13.93%, 14.78%, 14.28%, 14.03% and 13.25%) toxins were also higher at same conditions. These optimized conditions can be used for major toxin gene expression studies of C. perfringens toxinotype B.

The current study was conducted to examine the possible advantages of Heydotis corymbosa (L.) Lam. extract nanogel as a perspective for enhanced permeation and extended skin deposition in psoriasis-like dermatitis. Optimised nanophytosomes (NPs) were embedded in a pluronic gel base to obtain nanogel and tested ex vivo (skin penetration and dermatokinetics) and in vivo. The optimised NPs had a spherical form and entrapment efficiency of 73.05 ± 1.45% with a nanosized and zeta potential of 86.11 nm and −10.40 mV, respectively. Structural evaluations confirmed encapsulation of the drug in the NPs. Topical administration of prepared nanogel to a rat model of psoriasis-like dermatitis revealed its specific in vivo anti-psoriatic efficacy in terms of drug activity compared to the control and other formulations. Nanogel had improved skin integrity and downregulation of inflammatory cytokines. These findings suggest that developed phytoconstituent-based nanogel has the potential to alleviate psoriasis-like dermatitis with better skin retention and effectiveness.

Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are enzymes that break down and reduce the level of the neurotransmitter acetylcholine (ACh). This can cause a variety of cognitive and neurological problems, including Alzheimer’s disease. Taxifolin is a natural phytochemical generally found in yew tree bark and has significant pharmacological properties, such as being anti-cancer, anti-inflammatory, and antioxidant. The binding affinity and inhibitory potency of taxifolin to these enzymes were evaluated through molecular docking and molecular dynamics simulations followed by the MMPBSA approach, and the results were significant. Taxifolin’s affinity for binding to the AChE–taxifolin complex was −8.85 kcal/mol, with an inhibition constant of 326.70 nM. It was observed to interact through hydrogen bonds. In contrast, the BChE–taxifolin complex binding energy was observed to be −7.42 kcal/mol, and it was significantly nearly equal to the standard inhibitor donepezil. The molecular dynamics and simulation signified the observed interactions of taxifolin with the studied enzymes. The MMPBSA total free energy of binding for AChE–taxifolin was −24.34 kcal/mol, while BChE–taxifolin was −16.14 kcal/mol. The present research suggests that taxifolin has a strong ability to bind and inhibit AChE and BChE and could be used to manage neuron-associated problems; however, further research is required to explore taxifolin’s neurological therapeutic potential using animal models of Alzheimer’s disease.

The pervasive challenge of heavy metal pollution in water necessitates urgent interventions to mitigate chromium contamination. It poses significant threats to human health, aquatic ecosystems, and the environment. This research delves into the potential of date palm waste as a precursor of adsorbent for extraction of chromium from contaminated water, underscoring its potential in heavy metal remediation. Date palm waste, traditionally regarded as agricultural waste, underwent modification employing N-(3-chloro-2-hydroxypropyl) trimethylammonium chloride resulting in quaternized date palm waste (QDPW) as a prospective adsorbent for chromium extraction. The characterization techniques, including FTIR, SEM/EDX, BET, XRD, TGA/DTA, and element analysis, were employed to examine the structural and surface properties of both DPW and QDPW fibers. Optimal chromium uptake was achieved at a solution pH 6.5 over two hours, with a peak removal efficiency of 88% and an adsorption capacity of 22.26 mg/g. The equilibrium isotherms studies are consistent with Langmuir, Redlich-Peterson, and Sips isotherms, presenting a coefficient of determination (R2) of 0.989, 0.981, and 0.983, respectively. Moreover, the pseudo-second-order model was mostly aligned with the kinetics data with R2 value of 0.999. These findings showed that the quaternized date palm waste can be considered a good option to recycle the date palm waste and reduce the environmental threats in aqueous solution.

Globally, malignancies cause one out of six mortalities, which is a serious health problem. Cancer therapy has always been challenging, apart from major advances in immunotherapies, stem cell transplantation, targeted therapies, hormonal therapies, precision medicine, and palliative care, and traditional therapies such as surgery, radiation therapy, and chemotherapy. Natural products are integral to the development of innovative anticancer drugs in cancer research, offering the scientific community the possibility of exploring novel natural compounds against cancers. The role of natural products like Vincristine and Vinblastine has been thoroughly implicated in the management of leukemia and Hodgkin’s disease. The computational method is the initial key approach in drug discovery, among various approaches. This review investigates the synergy between natural products and computational techniques, and highlights their significance in the drug discovery process. The transition from computational to experimental validation has been highlighted through in vitro and in vivo studies, with examples such as betulinic acid and withaferin A. The path toward therapeutic applications have been demonstrated through clinical studies of compounds such as silvestrol and artemisinin, from preclinical investigations to clinical trials. This article also addresses the challenges and limitations in the development of natural products as potential anti-cancer drugs. Moreover, the integration of deep learning and artificial intelligence with traditional computational drug discovery methods may be useful for enhancing the anticancer potential of natural products.

Begomoviruses are contagious and severely affect commercially important fiber and food crops. Cotton leaf curl Multan virus (CLCuMuV) is one of the most dominant specie of Begomovirus and a major constraint on cotton yield in Pakistan. Currently, the field of plant genome editing is being revolutionized by the CRISPR/Cas system applications such as base editing, prime editing and CRISPR based gene drives. CRISPR/Cas9 system has successfully been used against biotic and abiotic plant stresses with proof-of-concept studies in both model and crop plants. CRISPR/Cas12 and CRISPR/Cas13 have recently been applied in plant sciences for basic and applied research. In this study, we used a novel approach, multiplexed crRNA-based Cas12a toolbox to target the different ORFs of the CLCuMuV genome at multiple sites simultaneously. This method successfully eliminated the symptoms of CLCuMuV in Nicotiana benthamiana and Nicotiana tabacum. Three individual crRNAs were designed from the CLCuMuV genome, targeting the specific sites of four different ORFs (C1, V1 and overlapping region of C2 and C3). The Cas12a-based construct Cas12a-MV was designed through Golden Gate three-way cloning for precise editing of CLCuMuV genome. Cas12a-MV construct was confirmed through whole genome sequencing using the primers Ubi-intron-F1 and M13-R1. Transient assays were performed in 4 weeks old Nicotiana benthamiana plants, through the agroinfiltration method. Sanger sequencing indicated that the Cas12a-MV constructs made a considerable mutations at the target sites of the viral genome. In addition, TIDE analysis of Sanger sequencing results showed the editing efficiency of crRNA1 (21.7%), crRNA2 (24.9%) and crRNA3 (55.6%). Furthermore, the Cas12a-MV construct was stably transformed into Nicotiana tabacum through the leaf disc method to evaluate the potential of transgenic plants against CLCuMuV. For transgene analysis, the DNA of transgenic plants of Nicotiana tabacum was subjected to PCR to amplify Cas12a genes with specific primers. Infectious clones were agro-inoculated in transgenic and non-transgenic plants (control) for the infectivity assay. The transgenic plants containing Cas12a-MV showed rare symptoms and remained healthy compared to control plants with severe symptoms. The transgenic plants containing Cas12a-MV showed a significant reduction in virus accumulation (0.05) as compared to control plants (1.0). The results demonstrated the potential use of the multiplex LbCas12a system to develop virus resistance in model and crop plants against begomoviruses.

Foot and Mouth Disease (FMD) is globally pandemic which badly affect the economics of livestock based countries like Pakistan. There are different types of Foot and Mouth Disease Virus (FMDV) among these types O is most prevalent in Pakistan. Recently Pakistan is producing approximately fifteen million doses of non-purified FMD vaccine against the demand of 160 million doses annually. More over the Pakistan is still striving for the development and optimization of concentration as well as purification of FMDV. The present project was designed to develop the technology for the purification of FMDV indigenously. The locally isolated and adapted FMDV type O virus was propagated on adherent culture of BHK-21cells to get final volume of virus one liter. This virus suspension was concentrated by peggylation as well as ultra-filtration method. The purification and quantification of concentrated virus was done by size exclusion chromatography. The results showed that peggylation is better method of concentration up to 603.75 µg/ml with 82.80 % recovery rate than ultra-filtration with 43.90 % followed by chromatography for purification. The PD50 was calculated in bovines at 24, 12, 6, 3 and 1.5 µg of FMDV Ag/dose and it revealed that antigen load of 1.98 µg is the dose, where the 50 % of inoculated animals showed the protective antibody level based upon percent inhibition through antibody detecting ELISA. According to the British pharmacopeia, the vaccine should contain 3PD50 which found equivalent to our findings about 6 µg/dose. The group of animal injected with 6/dose (3.23PD50) showed protective titer up to 20th week post priming.

The current research was conducted to investigate and compare the role of functional foods such as probiotics and prebiotics alone or in combinations on the markers of liver damage, oxidative stress markers and inflammatory markers in the nonalcoholic fatty liver disease (NAFLD) induced rats. According to the results, G3, G4 and G6 gained significant weight at the end of the study. Moreover, the weights of different organs (liver, Spleen and kidney) revealed that G1 (control negative) fed on a standard diet gained the lowest liver weight, G2 (control positive) fed on high fat diet (HFD) gained maximum weight (9.63 ± 0.46 g). In addition, the symbiotic group (G8) lowered the liver organ weight (6.77 ± 0.20 g), indicating that HFD induced fat deposition in body and other body organs. Furthermore, results of serum liver function markers were concerned, Maximum reduction of Alanine aminotransferase (ALT) was found among the G8 (Probiotics, GOS and Inulin) and G4 (GOS). Aspartate aminotransferase (AST) was significantly reduced among all the treatments compared to the control positive. Alkaline phosphate (ALP) was reduced among G5, G6 and G7. Gamma-glutamyl transferase (GGT) remained unaffected during the trial. As far as the results of tumor necrotic factor (TNF) were concerned, probiotics and GOS groups had a significant reduction in the levels of TNF after 19 weeks of study. Upon the prebiotics and probiotics therapy, a strong association was found with the hepatic inflammation at various degrees and hepatic and portal inflammation was reduced among the symbiotic groups significantly (Chi-square value = 49.04**; P-value = 0.000; ** = Highly significant (P

The present study focused on evaluating the influence of ultrasonic-assisted pasteurization (UAP) on the quality and shelf stability of jambul squash. Squash was analyzed for physicochemical characteristics during a storage period of 4 months. There was a significant effect (p

Abstract Food is the basic necessity for life that always motivated man for its preservation and making it available for an extended period. Food scientists always tried to preserve it with minimum deterioration in quality by employing and investigating innovative preservation techniques. The food sector always remained in search of eco‐friendly and sustainable solutions to tackle food safety challenges. Green technologies (ozone, pulsed electric field, ohmic heating, photosensitization, ultraviolet radiations, high‐pressure processing, ultrasonic, nanotechnology) are in high demand owing to their eco‐friendly, rapid, efficient, and effective nature in controlling microbes with a negligible residual impact on food quality during processing. The use of green technologies would be a desirable substitute for conventionally available preservation techniques. This paper discusses different food preservation techniques with special reference to green technologies to minimize the deleterious impact on the environment and employs these innovative technologies to play role in enhancing the food safety.

Calotropis procera is a latex-producing plant with plenty of pharmacologically active compounds. The principal motivation behind this study was to separate and characterize laticifer proteins to check their antimicrobial potential. Laticifer proteins were separated by gel filtration chromatography (GFC) and investigated using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE). The SDS-PAGE assay detected proteins of molecular weights of 10 to 30 kDa but most of them were in the range of 25 to 30 kDa. The soluble laticifer proteins (SLPs) were tested against Gram-positive bacteria i.e., Streptococcus pyogenes and Staphylococcus aureus whereas Escherichia coli and Pseudomonas aeruginosa were tested as Gram-negative bacteria, we determined a profound anti-bacterial activity of these proteins. In addition, SLPs were also investigated against Candida albicans via the agar disc diffusion method which also showed significant anti-fungal activity. SLP exhibited antibacterial activity against P. aeruginosa, E. coli, and S. aureus with a minimum inhibitory concentration (MIC) of 2.5 mg/mL for each, while MIC was found at 0.625 mg/mL for S. pyogenes and 1.25 mg/mL for C. albicans. Moreover, enzymatic activity evaluation of SLP showed the proteolytic nature of these proteins, and this proteolytic activity was greatly enhanced after reduction which might be due to the presence of cysteine residues in the protein structure. The activity of the SLPs obtained from the latex of C. procera can be associated with the involvement of enzymes either proteases or, protease inhibitors and/or peptides.

Background: Heart failure (HF) is an important adverse outcome of diabetes mellitus (DM) with high rates of mortality and HF-related hospitalizations. The risk of HF is 2 times higher in patients with DM compared to those without DM. Due to under-recognition and underdiagnoses, HF is often a neglected outcome in the diabetic population. There is a dearth of data regarding the true prevalence of HF and the management protocols for diabetic patients at risk of HF in the UAE and the Middle East. This lacuna in the information has led to the inception of this “call to action” paper, which identifies the gaps in the true prevalence of HF and describes the importance of early diagnosis and appropriate management of HF in the Middle East. Methodology: An advisory board meeting was convened and a group of key opinion leaders and experts in cardiology and endocrinology assembled to describe the prevalence, diagnosis, and management of HF in diabetes patients and to present a “call to action” in the UAE and Middle East scenario. After the group discussion, key expert opinions were formulated and “call to action” recommendations were proposed. Conclusion: This “call to action” is mainly based on the available evidence from the literature and the experts’ clinical experience. Based on the new evidence from various cardiovascular outcome trials, the “call to action” highlights a series of collaborative learning regarding the role of newer antidiabetic therapies like sodium–glucose cotransporter-2 inhibitors in the prevention and management of HF. This “call to action” intends to serve as a guide for physicians, including primary healthcare providers, in their management of diabetic patients with HF.

Aedes aegypti is a principal vector for several viruses including dengue virus, chikungunya virus and zika virus. Economic burden of mosquito-borne diseases, relative failure of traditional control strategies and the resistance development against insecticides enforces towards genetic manipulation of Ae. aegypti. Hence, a key gene doublesex (Aedsx) which regulate sex differentiation and alternatively splices to form male and female specific transcripts (AedsxM and AedsxF). CRISPR/Cas9 technique was employed to sex specifically disrupt the female-specific isoforms, AedsxF1 and AedsxF2, both of which were shown to be expressed only in female mosquitoes. Targeting of dsxF at the developmental stage has resulted in various phenotypic anomalies of adult females. The rate of adult mutation phenotype was recorded between 29 and 37% along with anomalies of wing size, proboscis length and reduction in the sizes of pre-blood-meal and after blood-meal ovaries in dsxF1 and dsxF2 microinjected groups, respectively. These findings can be correlated with reduced fecundity rate of Go female, where AedsxF1 and AedsxF2 groups showed reduction rate in range of 23–31%. Furthermore, hatching inhibition rate of 28 to 36% was also observed in G1 generation when compared to the wildtype. Overall, these results demonstrated that AedsxF disruption has resulted in multiple female traits disruption including decreased fertility of the female that could directly or indirectly associated with reproduction and its disease transmitting abilities. All these findings suggesting that CRISPR works to alter the developmental pathways as predicted, and therefore this method potentially gives us the basis for the sex-ratio distortion system as genetic control approach for the management of this vector.

Energy hub planning and optimization considering various constraints is an emerging domain of research due to the scarcity of conventional energy resources. Strategic integration of multi-energy sources is a dominant region of research as it provides better efficiency and cost savings. Various uncertainties due to the changing electric, heating, and cooling load demand are increasing the complexity of the system. The paper investigates the effectiveness of Demand Side Management (DSM) programs by analyzing the operation of domestic Energy Hub. The paper proposes two stages for optimal energy hub planning and operation optimization and GAMS mathematical modeling is used to deduce the results. The paper provides the analysis of a two-stage optimization model using the approach of modeling uncertainties and constraints. The process of energy hub designing has been carried out by the help of PSO algorithm by considering continuous capacities of various components of energy hub. The research regarding optimal energy hub operation is modelled in GAMS and the approach of mixed-integer non-linear programming (MINLP) has been used to evaluate the results. The framework provides the energy hub model and also provided the verification of model effectiveness and efficiency