Development and Characterization of Selective CB2R Inverse Agonists as a Novel Anti-Inflammatory Therapy for Neuroinflammation – Mechanisms of Action in Murine and Human Microglia Models Provide Valuable Insights into Their Therapeutic Potential

Introduction: Recently, research has revealed the presence of cannabinoid receptor 2 (CB2R) in the central nervous system (CNS); since then, the receptor has emerged as a promising therapeutic target for treating multiple neurodegenerative diseases (NDD) such as Alzheimer’s (AD), Parkinson’s (PD), Human Immunodeficiency Virus (HIV) and Multiple Sclerosis (MS). The restricted and upregulated expression of CB2R in activated microglia and lack of psychotropic activity make this receptor an attractive target for discovering novel therapy to treat neuroinflammatory diseases for which there is currently no curative treatment. Methodology: A library of compounds were synthesized and evaluated in ACTOne and TANGO-bla functional assays for CB1 and CB2 activity in combination with affinity measurements for the CB1/2 receptors. Moreover, the compounds that demonstrated no activity at functional assays with high binding affinities to CB2R were further assessed in antagonist (competition) studies against the non-selective cannabinoid agonist CP 55,940. Compound 45 demonstrated CB2R inverse agonism at G-protein signaling with high potency, efficacy, and affinity in the binding and functional assays and was selected for further evaluation in murine and human microglia inflammatory models. On cell polarization assays were done to measure microglia M1 and M2 surface markers expression. Scratch assay and ROS measurements were performed to assess the effects of drug treatment on microglia migration and ROS production. Extensive signaling pathway studies were conducted to identify the downstream signaling mechanism for compound 45 and cytoskeleton staining was performed to evaluate the microglia morphological changes upon 45 treatment. A molecular docking study was utilized to explore ligand protein molecular interaction between 45 and CB2R. Additionally, the similarities and differences in the activity of compound 45, relative to standard CB2R agonist and inverse agonist, were examined in the aforementioned assays. Results: Compound 45 demonstrated the highest affinity and selectivity towards CB2R that translated to higher potency in CB2R ACTOne functional assay, whereas no functional activity was detected at CB1R. Moreover, testing compound 45 in the TANGO-bla assay revealed neutral antagonism at β-arrestins signaling, confirming the G-protein functional selectivity of this class of compounds. Treatment with compound 45 triggered a shift in murine and human microglia polarization from a pro-inflammatory (M1) to an anti-inflammatory (M2) phenotype after 24 hours of treatment. The M1 markers were significantly decreased and M2 markers significantly increased relative to LPS stimulus. These results were further confirmed using microscopic examination wherein compound 45 induced a remarkable transformation of microglia into ramified morphology with a small cell body and several elongated cell projections, indicating microglia adopted an anti-inflammatory M2 phenotype. Moreover, compound 45 significantly reduced cell migration, suppressed the M1 associated ROS production which causes neurotoxicity and cell damage. Additionally, the application of compound 45 significantly increased JNK, c-JUN, CREB, and p38 phosphorylation levels and decreased ERK1/2 and NF-κB representing a unique anti-inflammatory mechanism of CB2R inverse agonists in activated microglia. The molecular docking study of compound 45 revealed that the thiophene C ring constrained the toggle switch residue Trp 258 via multiple π-π stacking interactions that stabilized CB2R in the inactive state and led to 45 inverse agonism activity at CB2R. These results were distinctly different when compared to standard CB2R agonists and inverse agonists, suggesting further research is needed to explore more precisely the utility of CB2R based therapeutics in human CNS disorders. Conclusion: The results of these studies provided valuable information that aids in the development of new analogs of the biaryl-aryl scaffold for treating neuroinflammation in human neurodegenerative diseases

Stimulatory effects of Lycium shawii on human melanocyte proliferation, migration, and melanogenesis: In vitro and in silico studies

There is no first-line treatment for vitiligo, a skin disease characterized by a lack of melanin produced by the melanocytes, resulting in an urgent demand for new therapeutic drugs capable of stimulating melanocyte functions, including melanogenesis. In this study, traditional medicinal plant extracts were tested for cultured human melanocyte proliferation, migration, and melanogenesis using MTT, scratch wound-healing assays, transmission electron microscopy, immunofluorescence staining, and Western blot technology. Of the methanolic extracts, Lycium shawii L. (L. shawii) extract increased melanocyte proliferation at low concentrations and modulated melanocyte migration. At the lowest tested concentration (i.e., 7.8 μg/mL), the L. shawii methanolic extract promoted melanosome formation, maturation, and enhanced melanin production, which was associated with the upregulation of microphthalmia-associated transcription factor (MITF), tyrosinase, tyrosinase-related protein (TRP)-1 and TRP-2 melanogenesis-related proteins, and melanogenesis-related proteins. After the chemical analysis and L. shawii extract-derived metabolite identification, the in silico studies revealed the molecular interactions between Metabolite 5, identified as apigenin (4,5,6-trihydroxyflavone), and the copper active site of tyrosinase, predicting enhanced tyrosinase activity and subsequent melanin formation. In conclusion, L. shawii methanolic extract stimulates melanocyte functions, including melanin production, and its derivative Metabolite 5 enhances tyrosinase activity, suggesting further investigation of the L. shawii extract-derived Metabolite 5 as a potential natural drug for vitiligo treatment

Parental knowledge of RVS infection and attitude to infant immunization with monoclonal antibodies in western region, Saudi Arabia

Background: Human Respiratory Syncytial Virus (RSV) is a single-stranded (RNA) virus belongs to genus orthopneumovirus. It accounted as one of the main causes of lower respiratory tract infection in the pediatric age group and associated with their hospitalization and morbidity. Prophylactic monoclonal antibodies help in improvement and reduction of the serious complications resulting from the virus. Objective : The aim of this paper is to assess parental knowledge of RSV infection and attitudes to infant immunization with monoclonal antibodies. Method: A cross-sectional study conducted using online questionnaire from February 2023 to June 2023 in western region in the Kingdom of Saudi Arabia. A total of 606 of participants, which include any parents or caregiver who believe in childhood vaccination in western region of Saudi Arabia with exclusion of any parents who hesitate towards or refuse childhood vaccinations and outside western region. The statistical analysis done using IBM SPSS. Results: The study included 606 participants from the western province of Saudi Arabia, 218 (36%) were in the age group of 20-30 years, 383 (63.2%) had bachelor degrees And 77(12.7) works in healthcare, RSV was the least known childhood infectious agent (48.7%) of the participants never heard of it. 542 (89.4%) had a positive attitude toward childhood vaccinations in general and (51.2%) toward RSV vaccination with (75.2%) concerned about its safety.Conclusion: Despite the fact that practically all children were exposed to RSV, a sizable percentage of parents never heard of it. We propose that dependable healthcare professionals provide evidence-based information regarding RSV and its safety, effectiveness, and duration of protection against RSV for parents

Antimicrobial resistance among migrants in Europe: a systematic review and meta-analysis

BACKGROUND: Rates of antimicrobial resistance (AMR) are rising globally and there is concern that increased migration is contributing to the burden of antibiotic resistance in Europe. However, the effect of migration on the burden of AMR in Europe has not yet been comprehensively examined. Therefore, we did a systematic review and meta-analysis to identify and synthesise data for AMR carriage or infection in migrants to Europe to examine differences in patterns of AMR across migrant groups and in different settings. METHODS: For this systematic review and meta-analysis, we searched MEDLINE, Embase, PubMed, and Scopus with no language restrictions from Jan 1, 2000, to Jan 18, 2017, for primary data from observational studies reporting antibacterial resistance in common bacterial pathogens among migrants to 21 European Union-15 and European Economic Area countries. To be eligible for inclusion, studies had to report data on carriage or infection with laboratory-confirmed antibiotic-resistant organisms in migrant populations. We extracted data from eligible studies and assessed quality using piloted, standardised forms. We did not examine drug resistance in tuberculosis and excluded articles solely reporting on this parameter. We also excluded articles in which migrant status was determined by ethnicity, country of birth of participants' parents, or was not defined, and articles in which data were not disaggregated by migrant status. Outcomes were carriage of or infection with antibiotic-resistant organisms. We used random-effects models to calculate the pooled prevalence of each outcome. The study protocol is registered with PROSPERO, number CRD42016043681. FINDINGS: We identified 2274 articles, of which 23 observational studies reporting on antibiotic resistance in 2319 migrants were included. The pooled prevalence of any AMR carriage or AMR infection in migrants was 25·4% (95% CI 19·1-31·8; I2 =98%), including meticillin-resistant Staphylococcus aureus (7·8%, 4·8-10·7; I2 =92%) and antibiotic-resistant Gram-negative bacteria (27·2%, 17·6-36·8; I2 =94%). The pooled prevalence of any AMR carriage or infection was higher in refugees and asylum seekers (33·0%, 18·3-47·6; I2 =98%) than in other migrant groups (6·6%, 1·8-11·3; I2 =92%). The pooled prevalence of antibiotic-resistant organisms was slightly higher in high-migrant community settings (33·1%, 11·1-55·1; I2 =96%) than in migrants in hospitals (24·3%, 16·1-32·6; I2 =98%). We did not find evidence of high rates of transmission of AMR from migrant to host populations. INTERPRETATION: Migrants are exposed to conditions favouring the emergence of drug resistance during transit and in host countries in Europe. Increased antibiotic resistance among refugees and asylum seekers and in high-migrant community settings (such as refugee camps and detention facilities) highlights the need for improved living conditions, access to health care, and initiatives to facilitate detection of and appropriate high-quality treatment for antibiotic-resistant infections during transit and in host countries. Protocols for the prevention and control of infection and for antibiotic surveillance need to be integrated in all aspects of health care, which should be accessible for all migrant groups, and should target determinants of AMR before, during, and after migration. FUNDING: UK National Institute for Health Research Imperial Biomedical Research Centre, Imperial College Healthcare Charity, the Wellcome Trust, and UK National Institute for Health Research Health Protection Research Unit in Healthcare-associated Infections and Antimictobial Resistance at Imperial College London

Surgical site infection after gastrointestinal surgery in high-income, middle-income, and low-income countries: a prospective, international, multicentre cohort study

Background: Surgical site infection (SSI) is one of the most common infections associated with health care, but its importance as a global health priority is not fully understood. We quantified the burden of SSI after gastrointestinal surgery in countries in all parts of the world. Methods: This international, prospective, multicentre cohort study included consecutive patients undergoing elective or emergency gastrointestinal resection within 2-week time periods at any health-care facility in any country. Countries with participating centres were stratified into high-income, middle-income, and low-income groups according to the UN's Human Development Index (HDI). Data variables from the GlobalSurg 1 study and other studies that have been found to affect the likelihood of SSI were entered into risk adjustment models. The primary outcome measure was the 30-day SSI incidence (defined by US Centers for Disease Control and Prevention criteria for superficial and deep incisional SSI). Relationships with explanatory variables were examined using Bayesian multilevel logistic regression models. This trial is registered with ClinicalTrials.gov, number NCT02662231. Findings: Between Jan 4, 2016, and July 31, 2016, 13 265 records were submitted for analysis. 12 539 patients from 343 hospitals in 66 countries were included. 7339 (58·5%) patient were from high-HDI countries (193 hospitals in 30 countries), 3918 (31·2%) patients were from middle-HDI countries (82 hospitals in 18 countries), and 1282 (10·2%) patients were from low-HDI countries (68 hospitals in 18 countries). In total, 1538 (12·3%) patients had SSI within 30 days of surgery. The incidence of SSI varied between countries with high (691 [9·4%] of 7339 patients), middle (549 [14·0%] of 3918 patients), and low (298 [23·2%] of 1282) HDI (p < 0·001). The highest SSI incidence in each HDI group was after dirty surgery (102 [17·8%] of 574 patients in high-HDI countries; 74 [31·4%] of 236 patients in middle-HDI countries; 72 [39·8%] of 181 patients in low-HDI countries). Following risk factor adjustment, patients in low-HDI countries were at greatest risk of SSI (adjusted odds ratio 1·60, 95% credible interval 1·05–2·37; p=0·030). 132 (21·6%) of 610 patients with an SSI and a microbiology culture result had an infection that was resistant to the prophylactic antibiotic used. Resistant infections were detected in 49 (16·6%) of 295 patients in high-HDI countries, in 37 (19·8%) of 187 patients in middle-HDI countries, and in 46 (35·9%) of 128 patients in low-HDI countries (p < 0·001). Interpretation: Countries with a low HDI carry a disproportionately greater burden of SSI than countries with a middle or high HDI and might have higher rates of antibiotic resistance. In view of WHO recommendations on SSI prevention that highlight the absence of high-quality interventional research, urgent, pragmatic, randomised trials based in LMICs are needed to assess measures aiming to reduce this preventable complication

The experience of Saudi female students attending mixed gender ESL courses in Melbourne, Australia

This study explored the experience of Saudi female students attending mixed-gender English as a second language (ESL) courses in Melbourne, Australia and the factors that impacted their levels of engagement. A transcendental phenomenological research design was used to provide a thick description of the essence of the Saudi female experience in this learning environment. The participants included four Saudi female students currently attending ESL courses in Melbourne, Australia. Two phases—descriptive and interpretive—were employed in the data analysis to accurately capture the nature of the experience.<br> <br> This study revealed that, while all the participants held a positive attitude towards their experience, language proficiency level and marital status played crucial roles in their adaptation. Participants with high language skills reported smoother adaptation than students with low language skills. The married students tended to be more inhibited than single students by the attendance of male especially Saudi male. Working solely with males or participating in physical activities with males had the greatest impact on levels of engagement in classroom activities. These findings were broadly in line with the sparse literature available on this phenomenon. Therefore, future research suggestions and practical implications were offered

The Experience of Saudi Female Students Attending Mixed Gender ESL Courses in Melbourne, Australia.

This study explored the experience of Saudi female students attending mixed-gender English as a second language (ESL) courses in Melbourne, Australia and the factors that impacted their levels of engagement. A transcendental phenomenological research design was used to provide a thick description of the essence of the Saudi female experience in this learning environment. The participants included four Saudi female students currently attending ESL courses in Melbourne, Australia. Two phases—descriptive and interpretive—were employed in the data analysis to accurately capture the nature of the experience. This study revealed that, while all the participants held a positive attitude towards their experience, language proficiency level and marital status played crucial roles in their adaptation. Participants with high language skills reported smoother adaptation than students with low language skills. The married students tended to be more inhibited than single students by the attendance of male especially Saudi male. Working solely with males or participating in physical activities with males had the greatest impact on levels of engagement in classroom activities. These findings were broadly in line with the sparse literature available on this phenomenon. Therefore, future research suggestions and practical implications were offered

A critical review of cannabis in medicine and dentistry: a look back and the path forward.

INTRODUCTION: In the last two decades, our understanding of the therapeutic utility and medicinal properties of cannabis has greatly changed. This change has been accompanied by widespread cannabis use in various communities and different age groups, especially within the United States. With this increase, we should consider the potential effects of cannabis-hemp on general public health and how they could alter therapeutic outcomes. MATERIAL AND METHODS: The present investigation examined cannabis use for recreational and therapeutic use and a review of pertinent indexed literature was performed. The focused question evaluates "how cannabis or hemp products impact health parameters and do they provide potential therapeutic value in dentistry, and how do they interact with conventional medicines (drugs)." Indexed databases (PubMed/Medline, EMBASE) were searched without any time restrictions but language was restricted to English. RESULTS: The review highlights dental concerns of cannabis usage, the need to understand the endocannabinoid system (ECS), cannabinoid receptor system, its endogenous ligands, pharmacology, metabolism, current oral health, and medical dilemma to ascertain the detrimental or beneficial effects of using cannabis-hemp products. The pharmacological effects of pure cannabidiol (CBD) have been studied extensively while cannabis extracts can vary significantly and lack empirical studies. Several metabolic pathways are affected by cannabis use and could pose a potential drug interaction. The chronic use of cannabis is associated with health issues, but the therapeutic potential is multifold since there is a regulatory role of ECS in many pathologies. CONCLUSION: Current shortcomings in understanding the benefits of cannabis or hemp products are limited due to pharmacological and clinical effects not being predictable, while marketed products vary greatly in phytocompounds warrant further empirical investigation. Given the healthcare challenges to manage acute and chronic pain, this review highlights both cannabis and CBD-hemp extracts to help identify the therapeutic application for patient populations suffering from anxiety, inflammation, and dental pain

Giant cell lichenoid dermatitis within healed varicella zoster site

Key Clinical Message There are dermatoses that arise within healed zosteriform sites, such as granulomas annulare, acneiform eruptions, psoriasis, lichen planus, and giant cell lichenoid dermatitis “GCLD.” Nonetheless, graft‐versus‐host disease should be considered and ruled out, especially in patients post‐bone marrow transplant. Herein, we report a case of GCLD manifesting within healed zosteriform sites