Kallikrein directly interacts with and activates Factor IX, resulting in thrombin generation and fibrin formation independent of Factor XI.

Kallikrein (PKa), generated by activation of its precursor prekallikrein (PK), plays a role in the contact activation phase of coagulation and functions in the kallikrein-kinin system to generate bradykinin. The general dogma has been that the contribution of PKa to the coagulation cascade is dependent on its action on FXII. Recently this dogma has been challenged by studies in human plasma showing thrombin generation due to PKa activity on FIX and also by murine studies showing formation of FIXa-antithrombin complexes in FXI deficient mice. In this study, we demonstrate high-affinity binding interactions between PK(a) and FIX(a) using surface plasmon resonance and show that these interactions are likely to occur under physiological conditions. Furthermore, we directly demonstrate dose- and time-dependent cleavage of FIX by PKa in a purified system by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and chromogenic assays. By using normal pooled plasma and a range of coagulation factor-deficient plasmas, we show that this action of PKa on FIX not only results in thrombin generation, but also promotes fibrin formation in the absence of FXII or FXI. Comparison of the kinetics of either FXIa- or PKa-induced activation of FIX suggest that PKa could be a significant physiological activator of FIX. Our data indicate that the coagulation cascade needs to be redefined to indicate that PKa can directly activate FIX. The circumstances that drive PKa substrate specificity remain to be determined.

Elimination of fibrin γ-chain cross-linking by FXIIIa increases pulmonary embolism arising from murine inferior vena cava thrombi.

The onset of venous thromboembolism, including pulmonary embolism, represents a significant health burden affecting more than 1 million people annually worldwide. Current treatment options are based on anticoagulation, which is suboptimal for preventing further embolic events. In order to develop better treatments for thromboembolism, we sought to understand the structural and mechanical properties of blood clots and how this influences embolism in vivo. We developed a murine model in which fibrin γ-chain cross-linking by activated Factor XIII is eliminated (FGG3X) and applied methods to study thromboembolism at whole-body and organ levels. We show that FGG3X mice have a normal phenotype, with overall coagulation parameters and platelet aggregation and function largely unaffected, except for total inhibition of fibrin γ-chain cross-linking. Elimination of fibrin γ-chain cross-linking resulted in thrombi with reduced strength that were prone to fragmentation. Analysis of embolism in vivo using Xtreme optical imaging and light sheet microscopy demonstrated that the elimination of fibrin γ-chain cross-linking resulted in increased embolization without affecting clot size or lysis. Our findings point to a central previously unrecognized role for fibrin γ-chain cross-linking in clot stability. They also indirectly indicate mechanistic targets for the prevention of thrombosis through selective modulation of fibrin α-chain but not γ-chain cross-linking by activated Factor XIII to reduce thrombus size and burden, while maintaining clot stability and preventing embolism.

Exploring the effectiveness of pharmacy curriculum in Saudi Arabia in developing leadership skills among pharmacy students from their perspective: A mixed-methods study.

Introduction: Leadership is an important component of pharmacy education. This study helps identify the strengths and limitations of the pharmacy curriculum in developing leadership skills among pharmacy students in Saudi Arabia and suggest how to improve the curriculum to better prepare them to become effective leaders in the healthcare profession. Methods: This study employed a mixed-methods research design with a sequential exploratory design. Phase I involved semi-structured interviews with student leaders of pharmacy student clubs/societies from different universities in Saudi Arabia. Interviews were recorded and transcribed verbatim. The written transcriptions were analyzed using thematic analysis. Phase II used a survey questionnaire to collect data from a larger sample of pharmacy students and interns from different universities in Saudi Arabia and the data is mainly presented as frequencies and percentages. Results: Eleven eligible participants were interviewed. Thematic analysis generated 288 codes which were categorized into 17 subthemes. These subthemes were further categorized into five overarching themes: Leadership development and acquisition; Skills and characteristics of effective leaders; Challenge and support for student leaders; Personal growth and benefits of leadership; Vision, goals and responsibilities of student leaders. The findings informed the development of the survey questionnaire which was completed by 484 students/interns. Mixed opinions were received regarding whether the pharmacy curriculum/program is helping or has helped the respondents develop leadership skills as well as whether the college has provided (or is providing) adequate support, resources or opportunities for the development of leadership skills. Eighty-eight per cent of the participants requested that the pharmacy curriculum in their college should include more courses or workshops focused on leadership development. They favoured several options from the listed topics to be included in these courses or workshops of which effective communication was the most prominent. Conclusion: This study highlights that pharmacy curriculum/colleges in Saudi Arabia promote essential leadership skills through various pedagogical approaches and support mechanisms to some extent. However, the students identified that the pharmacy curriculum needs to be revised in order to better prepare them for leadership positions.

COVID-19 dynamics and immune response: Linking within-host and between-host dynamics.

The global impact of COVID-19 has led to the development of numerous mathematical models to understand and control the pandemic. However, these models have not fully captured how the disease's dynamics are influenced by both within-host and between-host factors. To address this, a new mathematical model is proposed that links these dynamics and incorporates immune response. The model is compartmentalized with a fractional derivative in the sense of Caputo-Fabrizio, and its properties are studied to show a unique solution. Parameter estimation is carried out by fitting real-life data, and sensitivity analysis is conducted using various methods. The model is then numerically implemented to demonstrate how the dynamics within infected hosts drive human-to-human transmission, and various intervention strategies are compared based on the percentage of averted deaths. The simulations suggest that a combination of medication to boost the immune system, prevent infected cells from producing the virus, and adherence to COVID-19 protocols is necessary to control the spread of the virus since no single intervention strategy is sufficient.

Situational judgment using ethical reasoning in Saudi undergraduate pharmacy students.

INTRODUCTION: There is a paramount need for moral development for pharmacists and pharmacy students to practice the patient-centered profession. We aimed to explore the current situational judgment utilizing ethical reasoning among undergraduate pharmacy students. METHODS: A set of ten ethical dilemmas, representing potential real-life situations that the students come across in the university and may face in the future as a pharmacist were developed by a team of students, academic staff, and stakeholders. These ethical dilemmas were validated, checked for accuracy, and piloted. An online questionnaire was created consisting of these ten scenarios as open-ended questions and administered to fourth year and fifth year PharmD students in a public university located at the city of Mecca, Saudi Arabia, asking them how they would react in that situation. Responses of the participants were analyzed using thematic analysis independently by four researchers and inter-rater agreement were achieved through consensus. RESULTS: Out of 205, 186 students completed the questionnaire with a response rate of 90.7%. Analysis and resulted in the generation of 32 codes, which were then categorized into seven overarching themes: student engagement, social and professional responsibility, academic integrity, legal obligation, moral obligation, signposting, and moral engagement and patient safety. CONCLUSIONS: Undergraduate pharmacy students experience complex state of mind in connection with ethical reasoning. The participants' situational judgment were driven by cultural norm, authority, and responsibility. Student engagement is also affected by the state of mind and feelings of mutual trust, perceived cultural influence and peer pressure. The students were prone to seek help from university administrators or teachers when faced with situations in which they were helpless.

Prescribing patterns of antidiabetics in type 2 diabetes and factors affecting them.

Introduction: There are limited studies that address the use of newer antidiabetics by prescribers in Saudi Arabia and their adoption of the newer diabetes guidelines. The primary aim of our study was to explore the prescribing patterns of first- and second-line agents for the management of type 2 diabetes and the factors affecting them. Methods: A cross-sectional self-administered survey was designed to collect responses from the prescribers in the Makkah Region of Saudi Arabia from 16 February through 16 June 2021. The questionnaire was developed in the English language, piloted, and tested for validity. Participants were recruited on a convenience sampling basis, and the data were collected by visiting them in-person. Results: A total of 103 prescribers responded to the survey. The majority of the respondents were male (69%), Saudi nationals (54%), practicing in Makkah city (62%), working in the government sector (62%), and had the prescribing experience of 1-5 years (37%). Metformin (95%) was the respondents' most preferred first-line drug of choice, followed by sulfonylureas (5%). When the first-line was contraindicated, sulfonylureas (30%) were considered the most preferred drug of choice, followed by DPP4 inhibitors (25%) and SGLT2 inhibitors (16%), respectively. When the first-line was not sufficient to control diabetes, sulfonylureas (32%) and DPP4 inhibitors (31%) were considered the most preferred drugs of choice as an add-on therapy. SGLT2 inhibitors (31%) and metformin (24%) were considered the preferred first-line choices if the patient had an established cardiovascular disease. Conclusion: Sulfonylureas were found to be the most considered choice as second-line therapy. There is a need to provide targeted education to the prescribers related to the newer diabetes guidelines in order to promote the use of more evidence-based and safer antidiabetics.

Extracellular product of Pseudomonas aeruginosa in growth medium is involved in the pro-inflammatory cytokine response of human oral epithelial cells in vitro.

BACKGROUND: Epithelial cells are the first barrier to any microbial invasion. Finding a safe and affordable substance to stimulate the innate immune response of epithelial cells is one of the main challenges immunologists and vaccine manufacturers are facing. OBJECTIVES: This study aimed to show the comparative effect of sterile bacterial secretion (SBS) and Pseudomonas aeruginosa bacterial cell isolates obtained from burn wound infections on the ability of human epithelial cells (HECs) to produce interleukin (IL)-1ß and tumor necrosis factor alpha (TNF-α) in vitro. MATERIAL AND METHODS: The HEC cultures were exposed to P. aeruginosa 8 (Pa 8), Pa 2 and Pa 1 bacterial cells (isolated from burn wound infections). The other 3 groups of HECs were exposed to 50 µL of sterile, endotoxin-free SBS of Pa 8, Pa 2 and Pa 1. The time course of changes in IL-1ß mRNA, TNF-α mRNA, IL-1ß, and TNF-α was examined. RESULTS: Moderate (p < 0.05) elevations of IL-1ß mRNA in HECs and IL-1ß protein in the supernatant of the HEC culture were observed following exposure to SBS of Pa 8, Pa 2 and Pa 1 at most time points. High elevation (p < 0.05) of IL-1ß was seen in the supernatant of the HEC culture that was exposed to bacterial cells (Pa 8, Pa 2 and Pa 1). Similar results were found when TNF-α mRNA was measured in HECs and TNF-α in the supernatant of the HEC cultures after exposure to bacterial cells (Pa 8, Pa 2 and Pa 1) and the SBS of Pa 8, Pa 2 and Pa 1. CONCLUSIONS: This is the first time that the capacity of SBS to generate epithelial cell pro-inflammatory cytokines in vitro has been shown. In other words, SBS enhanced a nonspecific immune response, which opens the door to the possibility of using SBS from P. aeruginosa as an adjuvant in the future.

Neutrophil/lymphocyte ratio-A marker of COVID-19 pneumonia severity.

AIM: To determine the efficacy of neutrophil/lymphocyte ratio (NLR) as a marker of the severity of COVID-19 pneumonia in the South-Asian population. METHODS: This was a prospective, cross-sectional, analytic study conducted at HDU/ICU of District Headquarter Hospital, Faisalabad, Pakistan, from May through July 2020. Sixty-three eligible patients, admitted to the HDU/ICU, were prospectively enrolled in the study. Their NLR, C-reactive protein, serum albumin and serum fibrinogen were measured. Patients' demographic characteristics, comorbidities, clinical manifestations of COVID-19 infection, medication use and history of lung malignancy were retrieved from their medical history. Patients were categorised into either a general group (with mild COVID-19) or a heavy group (with moderate to severe COVID-19). RESULTS: There were significant differences between the two groups in diabetes prevalence, NLR, C-reactive protein and serum albumin. NLR and C-reactive protein were positively correlated (P < .001, P = .04, respectively) whereas serum albumin was negatively correlated (P = .009) with severe COVID-19. NLR was found to be an independent risk factor for severe COVID-19 pneumonia in the heavy group (OR = 1.264, 95% CI: 1.046~1.526, P = .015). The calculated AUC using ROC for NLR was 0.831, with an optimal limit of 4.795, sensitivity of 0.83 and specificity of 0.75, which is highly suggestive of NLR being a marker for the early detection of deteriorating severe COVID-19 infection. CONCLUSION: NLR can be used as an early warning signal for deteriorating severe COVID-19 infection and can provide an objective basis for early identification and management of severe COVID-19 pneumonia.

Implementation of a telemedicine service during COVID-19 pandemic in Pakistan.

AIM: In this feasibility study, we aimed to implement a pilot telemedicine service at our healthcare facility in Faisalabad, Pakistan, and describe our experience. METHODS: Telemedicine service was established by Faisalabad Medical University at two of its affiliated locations: (i) a 24-hour COVID-19 Telephone Helpline was established at the Director Emergency Office, Allied Hospital, and District Headquarter (DHQ) Hospital. (ii) A Telemedicine Clinic comprising consultants and postgraduate residents from different specialties was established at Chief Office, Allied Hospital. The data related to the number and categories of calls and advice provided were collected from 27th March 2020 to 31st July 2020. RESULTS: A total of 4582 calls were received, at both locations, during the study period, out of which 2325 callers (51%) were male, and 2257 (49%) were females. At Allied Hospital, 172 patients were advised accordingly for their complaints, whereas, at DHQ Hospital, 320 patients were advised accordingly for their complaints. At the Telemedicine Clinic only, a total of 2436 calls were received during the study period, 1474 (60%) callers were male, and 962 (40%) were female. The majority of the calls were received by medicine (43%), dermatology (21%), and paediatrics (11%) specialties, respectively. CONCLUSION: Despite some limitations, the benefits of telemedicine in this COVID-19 era are enormous and it is feasible to implement telemedicine services in developing countries. The developing countries must invest in the internet and technology access to facilitate telemedicine and other e-health services for not only curbing this pandemic but also to promote a more efficient healthcare system after the pandemic.

Clinical characteristics and outcomes of patients with COVID-19 pneumonia admitted to an intensive care unit in Faisalabad, Pakistan.

AIM: To describe the clinical characteristics and outcomes of adult patients with severe COVID-19, with the exploration of risk factors for mortality in the hospital. METHODS: This study included 20 adult patients diagnosed with COVID-19 admitted to the ICU of DHQ Hospital, Faisalabad (Pakistan). Patients were categorised into the survival group and the death group according to the outcome. We retrieved demographics, clinical manifestations and signs, laboratory indicators, treatment measures and clinical outcomes from the medical record, and summarised the clinical characteristics and outcomes of these patients. RESULTS: The average age of patients was 70 ± 12 years, of which 40% were male. They were admitted to the ICU 11 days after the onset of symptoms. The most common symptoms on admission were cough (19 cases, 95%), fatigue or myalgia (18 cases, 90%), fever (17 cases, 85%) and dyspnoea (16 cases, 80%). Eleven (55%) patients had underlying diseases, of which hypertension was the most common (11 cases, 55%), followed by cardiovascular disease (4 cases, 20%) and diabetes (3 cases, 15%). Six patients (30%) received invasive mechanical ventilation and continuous renal replacement therapy and eventually died. Acute heart injury was the most common complication (19 cases, 95%). Ten (50%) patients died between 2 and 19 days after admission to the ICU. Compared with dead patients, the average body weight of surviving patients was lower (61.70 ± 2.36 vs 68.60 ± 7.15, P = .01), Glasgow Coma Scale score was higher (14.69 ± 0.70 vs 12.70 ± 2.45, P = .03), with fewer concurrent shocks (2 vs 10, P = .001) and acute respiratory distress syndrome (2 vs 10, P = .001). CONCLUSION: The mortality rate is high in critically ill patients with COVID-19. Lower Glasgow Coma Scale, higher body weight and decreased lymphocyte count appear to be potential risk factors for the death of patients with COVID-19 in the ICU.

What just happened? Impact of on-campus activities suspension on pharmacy education during COVID-19 lockdown - A students' perspective.

INTRODUCTION: COVID-19 pandemic induced lockdown, suspending all on-campus educational activities in the Kingdom of Saudi Arabia (KSA), compelling to continue the education online. We explored pharmacy students' perspective on its impact on their learning. METHODS: A Twitter chat was organized on three consecutive days, after final examinations, inviting all pharmacy students in KSA to participate. Day 1 chat included 11 questions regarding learning and assessment, Day 2 chat included six questions about online examinations and six questions about technology use, Day 3 chat included six questions related to lessons learnt from the learning experiences during the lockdown. The questions were validated and piloted before the chat. The responses were copied, reviewed to remove any confidential information, and thematically analyzed by two teams of research students independently. RESULTS: During the three-day chat, 790 responses were received in total. Thematic analysis generated 944 codes which were categorized into 43 subthemes. These subthemes were further categorized into six main themes: 'facilitators for online education', 'barriers for online education', 'online versus onsite education', 'role of technology in online education', 'suggestions for improving online education' and 'long-term impact of online education during lockdown'. Participants highlighted several facilitators and barriers which affected their education during the lockdown, compared online education with onsite education, and provided suggestions for improving online education based on their learning experiences during the lockdown. CONCLUSION: As COVID-19 pandemic and its repercussions are expected to last longer, pharmacy colleges and academic staff will find these findings useful to prepare for the coming years, ensuring pedagogical and accreditation standards.

Development and evaluation of clinical reasoning using 'think aloud' approach in pharmacy undergraduates - A mixed-methods study.

INTRODUCTION: Given the widespread use of clinical reasoning (CR) in the healthcare practice, it is essential to inculcate the CR practice in undergraduate pharmacy education which can not only facilitate their clinical education and clinical rotations but can also help them become better clinical pharmacists. There is very limited CR employed in the pharmacy curriculum and practice in the Middle East countries. This study aimed to develop and evaluate CR practice in pharmacy undergraduates in one college of pharmacy in Saudi Arabia. METHODS: We employed a mixed-methods methodology that included two phases. In Phase I, students were introduced to CR practice ('think aloud' method) and given geriatric clinical cases which they used in two sessions together with a tutor. This was followed by the writing of SOAP notes using the tutor feedback and completion of a survey that included a self-reflection about their experience of using the CR method. Phase II included face-to-face semi-structured interviews involving selected students that were recruited via convenience sampling to further explore the issues identified in Phase I of the study. RESULTS: Of the 155 students who completed the survey (response rate 94%), the majority of them agreed that CR using the 'think aloud' method was useful in gathering (92%) and interpreting (95%) relevant patient information, identifying medication-related problems (95%), exploring therapeutic options for the problem(s) (93%) and formulating a treatment plan for the patient (90%). Qualitative data analysis of the 12 interviews was consistent with these findings. Furthermore, it provided an insight into the challenges faced by the students in applying this CR method. CONCLUSIONS: Students found the practice of CR using the 'think aloud' method helpful in working through given cases and taking clinical decisions. This method can be widely employed in pharmacy education and practice.

Risk Factors of Mid-upper Arm Circumference (MUAC) Based Child Malnutrition in the Flood-affected Areas of Pakistan: A Cross-sectional Study.

Low- and middle-income countries are usually at high risk of malnutrition. Not only that but the prevalence of malnutrition is much higher. It is important to evaluate the determinants of malnutrition in flood-affected areas of Pakistan. The present study examined the prevalence and risk factors of MUAC-based child malnutrition in flood-hit regions of Khyber Pakhtunkhwa, Pakistan. Multi-stage sampling was employed to select 656 households. Finally, 298 children of 6-59 months were selected. MUAC, an independent anthropometric parameter, was used to investigate the nutritional status of children. An automated logistic regression model was used to identify the risk factors of MUAC-based malnutrition. The prevalence of MUAC-based malnutrition was found 46%, including 40.5% females and 52.1% males. More than 90% of people had improved water quality and soap hand washing facility. Almost 17% of respondents had no toilet facility. Through automated logistic model, child age, maternal age, family size, income level, mother education, water quality, toilet facility were the significant determinants (P < .05) of MUAC-based undernutrition in flood affecting the area. The findings suggest that MUAC-based malnutrition can be minimized in flood-hit areas by targeting the listed risk factors. Community-based awareness programs regarding guidance on nutrition might be a key to reducing malnutrition in the target areas.

Benzylaminoethyureido-Tailed Benzenesulfonamides: Design, Synthesis, Kinetic and X-ray Investigations on Human Carbonic Anhydrases.

A drug design strategy of carbonic anhydrase inhibitors (CAIs) belonging to sulfonamides incorporating ureidoethylaminobenzyl tails is presented. A variety of substitution patterns on the ring and the tails, located on para- or meta- positions with respect to the sulfonamide warheads were incorporated in the new compounds. Inhibition of human carbonic anhydrases (hCA) isoforms I, II, IX and XII, involving various pathologies, was assessed with the new compounds. Selective inhibitory profile towards hCA II was observed, the most active compounds being low nM inhibitors (KIs of 2.8-9.2 nM, respectively). Extensive X-ray crystallographic analysis of several sulfonamides in an adduct with hCA I allowed an in-depth understanding of their binding mode and to lay a detailed structure-activity relationship.

Assessing quality of life using WHOQOL-BREF: A cross-sectional insight among patients on warfarin in Malaysia.

BACKGROUND AND AIM: The effect of anticoagulation control on overall Health-Related Quality of Life (HRQoL) in patients taking warfarin in Malaysia has not been explored yet. Therefore, this study aimed to evaluate HRQoL among patients on warfarin in Malaysia. METHODS: HRQoL among patients on warfarin was measured using WHOQOL-BREF (World Health Organization Quality of life Assessment: Brief Version). Descriptive and inferential statistical analysis was done using Statistical Package for the Social Sciences (SPSS) version 22. RESULTS: Out of 319 patients, more female patients were observed than the males (n = 221, 69.3%, and n = 98, 30.7% respectively). Mean scores for the physical (61.14 ± 15.96), psychological (68.58 ± 16.11), social (63.55 ± 27.06) and environmental domains (62.78 ± 17.58) were observed. Statistically, a significant association was found between the comorbidities other than CVDs with health satisfaction (p = 0.002), physical (p = 0.001), psychological (p < 0.001), social (p = 0.003) and environmental domains of the WHOQOL-BREF. A weak positive correlation between overall health satisfaction and the social domain (r = 0.153) and a moderate positive correlation between the physical and environmental domains (r = 0.628) of the WHOQOL-BREF were observed. CONCLUSION: Patients had overall better perceived HRQoL scores in the psychological domain as compared with the rest of the domains of the WHOQOL-BREF. Age, gender, employment status, education level, the indication of use and duration of warfarin therapy associated with overall perceived HRQoL.

Evaluation of Antimicrobial Stewardship Programs (ASPs) and their perceived level of success at Makkah region hospitals, Kingdom of Saudi Arabia.

Antimicrobial stewardship programs (ASPs) are collaborative efforts to optimize antimicrobial use in healthcare institutions through evidence-based quality improvement strategies. The general administration of pharmaceutical care in the Saudi ministry of health (MOH) is putting outstanding efforts in implementing antimicrobial stewardship in Saudi health care settings. Several surveys have been conducted globally and reported many types of antimicrobial stewardship strategies in health institutions and their effectiveness. This study aims to identify ASPs in Makkah region hospitals and their perceived level of success. We administered a regional survey to explore current progress and issues related to the implementation of ASPs in Makkah region hospitals at the pharmacy level (n = 25). Among responding hospitals, 19 (76%) hospitals, the most commonly reported ASP were as following: formulary restrictions (90%) for broad-spectrum antimicrobials and use of prospective feedback on antimicrobial prescribing (68%), use of clinical guidelines and pathways (100%), and use of automatic stop orders (68%) to limit inappropriate antimicrobial therapy. The study outcomes will also be of pivotal importance to devise policies and strategies for antimicrobial stewardship implementation in other non-MOH settings in the Makkah region. Based on our results, all reported institutions have at least one antimicrobial stewardship program in a process with a high success rate. A multidisciplinary ASP approach, active involvement of drug & therapeutic committee, formulary restrictions, and availability of education & training of pharmacists and physicians on ASP are the primary elements for perceived successful antimicrobial stewardship programs in the Makkah region hospitals.

Extraction and characterization of high purity chitosan by rapid and simple techniques from mud crabs taken from Abbottabad.

Chitosan and chitosan based materials offer diverse applications in the field of biotechnology, nanotechnology, pharmaceuticals, environmental protection and tissue engineering due to their various biological and physicochemical properties. Major sources of chitosan are shrimps, crabs and lobsters. Properties of chitosan differ with the degree of deacetylation and the molecular weight. Researchers are investigating to produce high quality chitosan in cost effective and time efficient way which was the aim of present study. The exoskeleton of mud crabs, taken from Abbottabad, was demineralized with 2mol/dm3 H2SO4 solution for 4hour and then, deproteinized with 2mol/dm3 NaOH solution for 4hour at room temperature. Yield of crude chitin was 78% which was deacetylated with 55% NaOH solution at 110°C for 4hour to obtain chitosan. After precipitation, the yield of pure chitosan form the crab shell was 39%. The degree of deacetylation of chitosan was 92% measured by potentiometric titration and the molecular weight was 1.2×106g/mol (1200KD), determined by viscometric method. We concluded that a high quality chitosan can be produced at commercial level in Pakistan by rapid and simple techniques.

Anti-proliferative potential of cyclotetrapeptides from Bacillus velezensis RA5401 and their molecular docking on G-Protein-Coupled Receptors.

Elucidation of bioactive chemical compounds from rhizobacteria is highly utilized in pharmaceuticals and naturopathy, due to their health benefits to human and plants. In current study, four cyclopeptides along with one phenyl amide were isolated from the ethyl acetate extract of Bacillus velezensis sp. RA5401. Their structures were determined and characterized as cycle (L-prolyl-L-leucyl)2 (1), cyclo (L-prolyl-l-valine)2 (2), cycle (L-phenylanalyl-L-propyl)2 (3), cyclo (D-pro-L-tyr-L-pro-L-tyr)2 (4) and N-(2-phenylethyl)acetamide (5) on the basis of electron spray ionization mass spectrometry (ESI-MS), nuclear magnetic resonance (NMR) techniques and comparison with the literature data. The five compounds have been isolated for the first time from this species. The effect of various concentrations of these compounds on the proliferation of MDA-MB-231 breast cancer cells was examined. It was found that 1 and 2 induced concentration-independent anti-proliferative effects, while 3, 4 and 5 inhibited cancer cell proliferation in a concentration-dependent manner. Furthermore, to determine the suitable binding targets of these compounds within cancer cell line, detailed target prediction and comparative molecular-docking studies were performed. The compounds 1 and 2 hit intracellular anti-cancer targets of proteases family, while compounds 3, 4 and 5 interacted with different membrane receptors of G-Protein-Coupled Receptors (GPCRs). In conclusion, the Bacillus velezensis RA5401 can be an ideal strain to produce anti-proliferative constituents at industrial scale.

Factor XIII A-Subunit V34L Variant Affects Thrombus Cross-Linking in a Murine Model of Thrombosis.

OBJECTIVE: Factor XIII (FXIII) cross-links fibrin upon activation by thrombin. Activation involves cleavage at residue 37 by thrombin, releasing an activation peptide. A common polymorphism (valine to leucine variant at residue 34, V34L), located in the activation peptide, has been associated with increased activation rates and paradoxically a protective effect in cardiovascular disease. There is, currently, no data available on the effects of V34L from in vivo models of thrombosis. We examined the effect of FXIII V34L on clot formation and cross-linking in vivo. APPROACH AND RESULTS: We generated a panel of full-length recombinant human FXIII-A2 variants with amino acid substitutions in the activation peptide to investigate the effect of these variants on activation rate, and we used wild-type, V34L, and alanine to glycine variant at residue 33 variants to study the effects of varying FXIII activation rate on thrombus formation in a murine model of FeCl3 injury. FXIII activation assay showed that residues 29, 30, 33, and 34 play a critical role in thrombin interaction. Full-length recombinant human FXIII-A2 V34L has significant effects on clot formation, structure, and lysis in vitro, using turbidity assay. This variant influenced fibrin cross-linking but not size of the thrombus in vivo. CONCLUSIONS: Mutations in the activation peptide of full-length recombinant FXIII regulate activation rates by thrombin, and V34L influences in vivo thrombus formation by increased cross-linking of the clot.

Growing success with sustainability: The influence of green HRM, innovation, and competitive advantage on environmental performance in the manufacturing industry.

President Xi set a long-term mitigation target of carbon neutrality by 2060 in September 2020 to cut carbon emissions and improve environmental performance. As a result, the current study aims to investigate how green human resource management and green innovation affect environmental performance in pursuit of the ultimate objective of carbon neutrality. Furthermore, the research considers the potential mediating effect of a green competitive advantage to clarify the mechanisms by which GHRM and green innovation impact environmental performance. Data from 278 employees in the manufacturing industry of Heilongjiang Province were collected through a questionnaire, for which we used a quantitative random sampling technique. The study employed the Smart PLS-4 structural equation modeling technique and revealed that all three factors significantly influenced environmental performance: green HRM, innovation, and competitive advantage. Resource-based view (RBV) theory served as the theoretical foundation for the study. The study uniquely contributes to the literature on GHRM, green innovation, competitive advantage, and environmental performance.