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Nagla A El-Shitany,1 Amina M Bagher,2 Lenah Saeed Binmahfouz,2 Basma G Eid,2 Haifa Almukadi,2 Shaimaa M Badr-Eldin,3,4 Manal El-Hamamsy,5 Zuhair M Mohammedsaleh,6 Fayez M Saleh,7 Mohammed S Almuhayawi,8 Samar A Alghamdi,9 Rana A Arab,10 Soad S Ali,11,12 Steve Harakeh,12,13 Badrah S Alghamdi14,15 1Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tanta University, Tanta, Egypt; 2Department of Pharmacology and Toxicology, Faculty of Pharmacy, King Abdulaziz University, Jeddah, Saudi Arabia; 3Department of Pharmaceutics, Faculty of Pharmacy, King Abdulaziz University, Jeddah, Saudi Arabia; 4Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Cairo University, Cairo, Egypt; 5Department of Clinical Pharmacy, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt; 6Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, University of Tabuk, Tabuk, Saudi Arabia; 7Department of Medical Microbiology, Faculty of Medicine, University of Tabuk, Tabuk, Saudi Arabia; 8Department of Medical Microbiology/Parasitology and Molecular Microbiology Laboratory, King Abdulaziz University Hospital, Jeddah, Saudi Arabia; 9Department of Oral Biology, Faculty of Dentistry, King Abdulaziz University, Jeddah, Saudi Arabia; 10Medicine Program, Ibn Sina National College for Medical Studies, Jeddah, Saudi Arabia; 11Department of Histology, Faculty of Medicine, Assiut University, Assiut, Egypt; 12Yousef Abdullatif Jameel Scientific Chair of Prophetic Medicine Application, Faculty of Medicine (FM), King Abdulaziz University, Jeddah, Saudi Arabia; 13King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia; 14Department of Physiology, Neuroscience Unit, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia; 15Pre-Clinical Research Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi ArabiaCorrespondence: Nagla A El-Shitany, Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tanta University, Tanta, Egypt, Email nagla_fouad@yahoo.com; naglaa.fouad@pharm.tanta.edu.egBackground: Like other vaccines, Pfizer BioNTech’s COVID-19 vaccine efficacy against SARS-CoV-2 virus infections begins to decline within a few months after the 2nd dose. On August 12, 2021, the FDA allowed additional Pfizer BioNTch’s COVID-19 vaccine dose (3rd or booster dose) for individuals with weakened immunity. This study aimed to evaluate the short-term adverse reactions (ADRs) of the 2nd and the 3rd doses of the Pfizer BioNTech COVID-19 vaccine.Methods: Information for this study was collected by Google Form questionnaire (online survey). The results included responses from 442 people, the majority from Saudi Arabia.Results: The most common local ADRs following the 3rd dose were injection site pain, injection site hypersensitivity, and axillary lymph node swelling. The most common systemic ADRs were fatigue, muscle pain, bone pain, headache, and fever less than 38ºC. Less common systemic ADRs were shivering, fever more than 38ºC, nasal congestion and rhinorrhea, arrhythmia, cough, abdominal pain, chest tightness, nausea, diarrhea, vomiting, and tachypnea. Rare systemic ADRs were constipation, dizziness and vertigo, lack of concentration, sore throat, excessive hair loss, dysmenorrhea and heavy menstruation, and Bell’s palsy. Severe allergic reactions were reported by 2.6% of participants after the 2nd dose, compared with none after the 3rd dose. Nasal congestion and runny nose are more frequent after the 3rd dose. The ADRs of the 2nd and 3rd doses were significantly more prevalent in females. 12% of participants reported ADRs lasting more than one week after the 3rd dose compared to 5% after the 2nd dose. People ≤ 60 years were more affected by the vaccine ADRs.Conclusion: Most of the ADRs reported after the 3rd vaccine dose were consistent with the Pfizer vaccine information sheet and similar to the 2nd dose ADRs.Keywords: Pfizer-BioNTech COVID-19 vaccine, adverse drug reactions, booster dose, second dose, injection site pain, fatigue, online questionnaire

Nagla A El-Shitany,1,2 Steve Harakeh,3 Shaimaa M Badr-Eldin,4,5 Amina M Bagher,1 Basma Eid,1 Haifa Almukadi,1 Badrah S Alghamdi,6 Ahlam A Alahmadi,7 Nibal A Hassan,8 Nariman Sindi,9 Samar A Alghamdi,10 Hailah M Almohaimeed,11 Zuhair M Mohammedsaleh,12 Turki M Al-Shaikh,13 Mohammed S Almuhayawi,14 Soad S Ali,15 Manal El-Hamamsy16,17 1Department of Pharmacology and Toxicology, Faculty of Pharmacy, King Abdulaziz University, Jeddah, Saudi Arabia; 2Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tanta University, Tanta, Egypt; 3Special Infectious Agents Unit, King Fahd Medical Research Center, Yousef Abdullatif Jameel Chair of Prophetic Medicine Application, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia; 4Department of Pharmaceutics, Faculty of Pharmacy, King Abdulaziz University, Jeddah, Saudi Arabia; 5Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Cairo University, Cairo, Egypt; 6Department of Physiology, Neuroscience Unit, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia; 7Department of Biological Sciences, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia; 8Department of Biology, Faculty of Science, Taif University, Taif, Saudi Arabia; 9Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia; 10Department of Oral Biology, Faculty of Dentistry, King Abdulaziz University, Jeddah, Saudi Arabia; 11Department of Basic Science, Faculty of Medicine, Princess Nourah Bint Abdulrahman University, Riyadh, Saudi Arabia; 12Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, University of Tabuk, Tabuk, Saudi Arabia; 13Department of Biology, College of Science and Arts at Khulis, University of Jeddah, Jeddah, Saudi Arabia; 14Department of Microbiology and Medical Parasitology, Faculty of Medicine, King Abdulaziz University, Jeddah, 21589, Saudi Arabia; 15Department of Histology, Faculty of Medicine, Assiut University, Assiut, Egypt; 16Department of Pharmacy Practice, Faculty of Pharmacy, King Abdulaziz University, Jeddah, Saudi Arabia; 17Department of Clinical Pharmacy, Faculty of Pharmacy, Ain Shams University, Cairo, EgyptCorrespondence: Nagla A El-ShitanyDepartment of Pharmacology and Toxicology, Faculty of Pharmacy, King Abdulaziz University, Al-Sheekh Kamal Street, Alsulimaniah, Jeddah, Saudi ArabiaTel +966 545923440Email nagla_fouad@yahoo.comBackground: The Pfizer-BioNTech COVID-19 vaccine has recently received emergency approval from the US FDA. The mRNA technology was used to manufacture the Pfizer vaccine; however, as a pioneering technology that has never been used in the manufacture of vaccines, many people have concerns about the vaccine’s side effects. Thus, the current study aimed to track the short-term side effects of the vaccine.Methods: The information in this study was gathered by a Google Form-questionnaire (online survey). The results included the responses of 455 individuals, all of whom are Saudi Arabia inhabitants. Adverse effects of the vaccine were reported after the first and the second doses.Results: The most common symptoms were injection site pain, headaches, flu-like symptoms, fever, and tiredness. Less common side effects were a fast heartbeat, whole body aches, difficulty breathing, joint pain, chills, and drowsiness. Rare side effects include Bell’s palsy and lymph nodes swelling and tenderness. Flu-like symptoms were more common among those under 60 years of age, while injection site pain was more frequent among recipients who were 60 years and older. The study revealed a significant increase in the number of females who suffered from the vaccine side effects compared to males. Difficulty of breathing was more reported among recipients who had been previously infected with the coronavirus compared to those who had not been previously infected.Conclusion: Most of the side effects reported in this study were consistent with Pfizer’s fact sheet for recipients and caregivers. Further studies are required to determine the long-term side effects.Keywords: Pfizer-BioNTech COVID-19 vaccine, side effects, injection site pain, hypersensitivity, flu-like symptoms, online survey

Abstract Many notable applications have been described for magnetic nanoparticles in delivery of diverse drugs and bioactive compounds into cells, magnetofection for the treatment of cancer, photodynamic therapy, photothermal therapy, and magnetic particle imaging (MPI). In response to the growing demand for magnetic nanoparticles for drug delivery or biomedical imaging applications, more effective and eco-friendly methodologies are required for large-scale biosynthesis of this nanoparticles. The major challenge in the large-scale biomedical application of magnetic nanoparticles lies in its low efficiency and optimization of nanoparticle production can address this issue. In the current study, a prediction model is suggested by the fractional factorial designs. The present study aims to optimize culture media components for improved growth and iron uptake of this strain. The result of optimization for iron uptake by the strain ASFS1 is to increase the production of magnetic nanoparticles by this strain for biomedical applications in the future. In the present study, design of experiment method was used to probe the effects of some key medium components (yeast extract, tryptone, FeSO4, Na2-EDTA, and FeCl3) on Fe content in biomass and dried biomass of strain ASFS1. A 25−1 fractional factorial design showed that Na2-EDTA, FeCl3, yeast extract-tryptone interaction, and FeSO4-Na2-EDTA interaction were the most parameters on Fe content in biomass within the experimented levels (p

Nowadays nanoparticles (NPs) due to their multidimensional applications in enormous different fields, has become an exciting research topic. In particular, they could attract a noticeable interest as drug deliver with increased bioavailability, therapeutic efficacy and drug specificity. Epigenetic can be considered as a complex network of molecular mechanism which are engaged in gene expression and have a vital role in regulation of environmental effects on ethology of different disorders like neurological disorders, cancers and cardiovascular diseases. For many of them epigenetic therapy was proposed although its application accompanied with limitations, due to drug toxicity. In this review we evaluate two aspects to epigenetic in the field of NPs: firstly, the role of epigenetic in regulation of nanotoxicity and secondly application of NPs as potential carriers for epidrugs.

Background and purpose: Considering the importance of internationalization in medical education, adaptation of successful international and national models can help to promote it. This study aimed at explaining the achievement of the universities of medical sciences in internationalization of medical education in Iran. Materials and methods: This descriptive-adaptive study was conducted in the international relations office at Kerman University of Medical Sciences (KMU). At first, top three universities in England, Pakistan, Ireland, Cyprus, and Greece were selected based on The Times Higher Education World University Rankings. According to the land-use planning classification in Iran, KMU is the university to follow and assess the performance of higher education in these countries. Then, information about the policies of these universities in internationalization of higher education was obtained using their websites or by request. We also collected information of KMU as a representative of eight universities of medical sciences and its affiliated academic and research centers. Results: Data about the ownership of universities, student recruitment strategies, and internationalization policies of the universities were compared with those of KMU and the weaknesses and strengths of KMU were determined. Conclusion: The universities of medical sciences in Iran are suggested to revise their missions, goals, and performance and aim to practice internationalization of higher education as an important strategy in their university prospects.

Background: Despite some obstacles, internationalization of medical education can promote the educational, research and clinical level of universities.Objectives: The purpose of this study was to explore the opportunities, challenges, and solutions of the internationalization of higher medical education in Iran.Methods: The present qualitative study was conducted with Brown and Clark's thematic content analysis approach. Purposeful sampling was performed using the snowball strategy, and the data were collected using semi-structured interviews. The statistical population included Iranian students and professors residing in universities abroad, experts and key officials working in the Deputy of International Relations in Iran’s Ministry of Health and Medical Education, the Vice-Chancellors of the International Relation Units of the country’s universities of medical sciences, and foreign students studying in Iran’s universities of medical sciences. The data were gathered by conducting 17 interviews with eligible individuals following the principle of data saturation. Finally, the data were subjected to thematic analysis by MAXQDA-10 software.Results: The data extracted from the interviews were categorized into 268 codes, 29 subcategories, and six categories. The themes identified were "specifications of an international higher education center", "advantages of internationalization", and "necessary items and requirements" in the field of requirements and criteria, "weaknesses and challenges of internationalization" in the field of challenges, "operational solutions for addressing challenges" and "successful domestic and foreign experiences of the internationalization of higher medical education centers " in the field of solutions. The subthemes of "weaknesses and challenges at the level of the university" and "hardware specifications" attained the highest frequency among open codes.Conclusion: A need to be responsive to many present and future needs of our country necessities that the higher medical education system moves in parallel with the globalization process and multicultural societies. In this regard, officials should pay attention to the requirements, criteria, challenges, and solutions of the internationalization of higher medical education in Iran based on the evidence disclosed in this study.

BACKGROUND: The effects of chemical, biological, radiological, and nuclear (CBRN) incidents on human society can be irreparable. Preparing the health system for these incidents is essential. This study aims to identify obstacles to hospital preparedness against CBRN incidents and provide solutions. MATERIALS AND METHODS: This qualitative study was conducted using semi-structured interview method in 2022. The research community included experts in the fields of CBRN, and 17 persons were included in the study through purposive sampling. The interviews were collected by interview guide and recorded face-to-face and online and were analyzed using thematic content analysis method in MS Word 2016. RESULTS: The interviews’ analysis was classified into two main categories and 34 sub-categories. Some of the most important obstacles to the hospital preparedness against CBRN incidents were the lack of proper crisis management, the lack of specialist staff, the stress of employees, the lack of turnover and sufficient rest for employees, legal gaps, and so on. The main strategies were determining the type and extent of the risk factor, strong crisis management, the lack of parallelism, continuous monitoring, having a protocol and road map, appropriate training programs, having skilled personnel, rapid response of personnel, positive attitude of the staff, and the favorable condition of the building. CONCLUSION: The appointment of an expert in the field of CBRN and having a specialized unit, the existence of specialized and trained staff along with access to the required facilities, clear instructions, and intra-departmental and inter-departmental cooperation affect the readiness of hospitals against CBRN incidents.

Background and Aim: Chemotherapy is one of the most common methods available for the treatment of cancer. Resistance to anticancer drugs can result in failures in chemotherapy of cancers. Therefore, investigation is necessary for the discovery of new drugs. Homoisoflavonoids are well-known natural products that possess a wide range of pharmacological activities. Hence, synthesis and assessment of cytotoxicity of homoisoflavonoids containing morpholinoethoxy moiety and their molecular docking simulation were taken into consideration. Materials and Methods: Two 7-morpholinoethoxyhomoisoflavonoids synthesized via three steps including SN2 reaction of 7-hydroxychromanine with morpholino ethyl chloride hydrochloride in the basic media under reflux condition to produce intermediate 3. The second step included aldol condensation of the intermediate with suitable aldehydes in the presence of gaseous HCl in ethanol, and finally neutralization with NaOH 5%. The structures of the products were confirmed using IR, 13CNMR, and 1HNMR techniques. Cytotoxic effects of final products on HT-29 and 3T3 cell lines were assessed. To evaluate the binding mode of compounds with tubulin protein, docking simulation was performed by using MOE. Results: Two morpholinoethoxy homoisiflavonoids were prepared with moderate to good production yield. Cytotoxic evaluation of these compounds showed satisfactory results. The results of molecular docking simulation showed an acceptable binding energy of the interaction between the ligands with the tubulin protein binding site: (E)-7-(2-morpholinoethoxy)-3-(3,4,5-trimethoxybenzylidene) chroman-4-one= -7.3727 Kcal/mol and (E)-3-(4-methoxybenzylidene)-7-(2-morpholinoethoxy) chroman-4-one released -7.6838 Kcal/mol per mole of energy. Conclusion:The results showed that compound (E)-3-(4-methoxybenzylidene)-7-(2-morpholinoethoxy) chroman-4-one has higher cytotoxicity than (E)-7-(2-morpholinoethoxy)-3-(3,4,5-trimethoxybenzylidene) chroman-4-one against HT-29 and 3T3 cell lines. The in-silico results confirmed the results obtained from in vitro experiments and showed three interactions between ligand (E)-3-(4-methoxybenzylidene)-7-(2-morpholinoethoxy)chroman-4-one and αThr179, αThr145, and αTyr224 amino acids in tubulin active site.

Background and Objectives: Cell-immobilization is used to maintain microbial culture to produce metabolites in repeated-batch or continuous fermentations, thereby reducing the time and resources spent on delivering mass production of microbe. The technique also enables shortening of the detoxification phase and the amount of formaldehyde required due to low incidence of viable bacteria in the extract. Materials and Methods: A solution of sodium alginate containing Clostridium perfringens cells was dropped into stirring CaCl2 solution via a sterile syringe needle. Optimizations resulted in reasonably uniform beads containing C. perfringens. Beads were externally stabilized by poly L-lysine, followed by immersion in a solution of Na-alginate to coat them with a new layer of alginate forming an alginate-PLL-alginate cortex. Results: This study proved successful in immobilizing C. perfringens cells inside uniform alginate microspheres. Cell loading and cell propagation inside the beads were measured. The cell loaded beads were cultivable in liquid media producing 550 minimum lethal doses per milliliter (MLD/ml) in a 72 h. Conclusion: The research paved the way for further investigations to optimize and establish an efficient bacterial encapsulation method. Thus, it seems possible to produce toxins from beads engulfing C. perfringens on larger scales via repeated-batch or continuous fermentation processes.

The main indicators of higher education (HE) internationalization in the field of awarding degrees are the international development of disciplines and interdisciplinary sciences, new educational and learning methods, new and updated curricula, and their correct ways of sharing. This study aimed to examine the system of awarding degrees in health HE of Iran and the United Kingdom. This descriptive-comparative study focused on the field of medical sciences and its related disciplines. The vital information about the variables was collected by visiting the official websites of the UK universities and related or joint organizations. The related information to the Kerman University of Medical Sciences as a sample of Iran medical universities was obtained from the university’s Farabar system. All data extraction steps were performed manually. There were differences in the mechanism of setting up a new discipline and the process of students’ admission, diversity of degrees’ titles and curriculums, stability of disciplines over the time, creativity in creating competition between different disciplines, the reason for establishing a discipline and the requirements for certification and awarding of degrees in health sciences disciplines in Iran and the United Kingdom were described. Propelling of medical education in the health sciences area towards standard awarding degree systems can be responsible for the requirements of internationalization of higher education.

During the last decade, plant and microbial-derived metabolites have received growing attention as potential tools for pest management in agriculture. Pederin (C25H45NO9) is a vesicant toxin produced by Pseudomonas-like bacterial symbionts of rove beetles within the genus Paederus (Col: Staphylinidae). In this study, the toxicity of pederin to two stored product pests, Ephestia kuehniella Zeller (Lep: Pyralidae) and Tribolium confusum Jacquelin du Val (Col: Tenebrionidae) was evaluated using laboratory bioassays. Probit analysis estimated the median lethal concentrations of pederin as 1311.96 and 596.36ppm for E. kuehniella fourth larval instar and T. confusum adults, respectively. We also measured the activity of two major digestive enzymes (amylases and proteases) as well as three major detoxifying enzymes (P450s monooxygenases, glutathione S-transferases, and carboxyl esterases) in insects treated orally with pederin. Feeding on pederin resulted in significant decrease in the activity of amylolytic, proteolytic, and carboxyl esterase enzymes, but significant increase in the activity of P450s and glutathione S-transferases. Results of this study may highlight pederin as a novel source of pesticides with unique mode of action for use in pest management programs.

The cyclodextrin glucanotransferase are extracellular enzymes that can hydrolysis starch as a substrate to cyclodextrin. Encapsulation and immobilization technique of microbial cells within the polymeric network have widespread applications in the production of enzyme. Bacterial cells were isolated from soil by serial dilution method. The molecular identity was established using 16S rDNA sequence analysis, and the identified strain was immobilized in the synthesized composites. Physicochemical characteristics of the synthesized composites were determined by using field emission scanning electron microscopy, Malvern Zetasizer Nano ZS and FT-IR analyses. Enzyme activity was measured by phenolphthalein assay. Streptomyces griseobrunneus was identified according to the 16S rDNA sequence. FT-IR analyses indicated that there was an interaction between all of the elements. Scanning electron microscopy analysis showed a cross-section of composites without cells and cells attached to composites. Enzyme activity was 1109, 948 and 606 U/ml in alginate/chitosan/microcrystalline cellulose (MCC) sheets, agar/gelatin/MCC beads and free cells, respectively. It could be concluded that the polymeric immobilization was effective in improving enzyme production because of its excellent properties such as guarding the bacterial cells against inhibitors and toxins.

Steroid hormones in plasma play an important role in reproductive cycles of animals especially during the final maturation stages. Steroid hormones synchronize gonad developments depending to fish species reproductive strategies. The wild Cyprinid fish, Kutum (Rutilus frisii kutum) is an ecologically and economically important fish species which inhabit in southern coastline of the Caspian Sea in Iran. Over the past few decades, natural reproduction of this species dramatically impaired due to the urbanization, civilization close to the land and shallow water in south western of the Caspian Sea. Therefore annual sex steroid hormones and gonads development were measured to assess the annual reproductive biology of female Kutum. In this study for the first time, our aims were to determine the annual variations in sex steroid hormones; 17 β Estradiol (E2), Progesterone (P) and testosterone (T) and gonad development of female Kutum. Our results showed that plasma steroid levels in females manifested in two phases in annual reproductive cycle; the resting phase (June - February) being characterized with the lowest level of steroid hormones and the peak reproduction activity phase (March–May) with simultaneously a significant increase in level of E2, P and T in plasma. Interestingly, comparing with other Teleost fish species the baseline level of E2 in plasma of Kutum during the resting phase to some extent was also huge. Increase in concentration of plasma E2 was in accordance with an increase of gonadosomatic index during spawning season. Our results contribute to our knowledge about the reproductive biology of Kutum and calls further long-term investigation.

Abstract Relation between sexual maturity and levels of two main steroid hormones in gonads, 17- estradiol (E2) and testosterone (T) were studied by using histological and radioimmunoassay in female kutum Rutilus frisii kutum during spawning season of the southern Caspian Sea. The study was carried out from February to May 2008 using 105 migrated fish specimens catched from the River sefid-rood by various tools of catching including (Gillnet, Cast net, Seine net and Sheyl or Kulham). The results revealed that changes in plasma levels of gonadal steroids, (E2) and (T) were closely correlated with ovarian development and increased in Gonadosomatic index (GSI) (P

Objectives: Ibuprofen, a widely used non-steroidal anti-inflammatory (NSAID) for managing pain and inflammation in pediatric patients, is metabolized by the CYP2C8 enzyme. Studies suggest that the CYP2C8*2 , *3 , and *4 variations of the CYP2C8 gene diminish ibuprofen metabolism, increasing the risk of adverse reactions. The aim of this study was to determine the frequency of the CYP2C8*2 , *3 , and *4 alleles and genotypes in a pediatric population attending the King Abdulaziz University dental clinic and compare our findings to those of other populations., Methods: A cross-sectional study was conducted with 140 healthy Saudi children ages 6-12. Saliva samples were collected using Oragene™ DNA Sample Collection Kits and analyzed for polymorphisms using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)., Results: The study identified that CYP2C8*2 AA, AT, and TT genotypes occurred at frequencies of 87.86 %, 9.29 %, and 2.86 %, respectively. For CYP2C8*3 , AA, AG, and GG genotypes were found in 87.14 , 8.75, and 4.29 % of subjects, respectively. The CYP2C8*4 allele was less frequent, with CC and CG genotypes at 97.86 % and 2.14 %, respectively, and the GG genotype was absent. Allele frequencies for CYP2C8*2 , *3 , and *4 were 7.5 %, 8.57 %, and 1.07 %, respectively., Conclusions: Our findings reveal that the allelic frequencies for the CYP2C8 polymorphisms in the Saudi pediatric cohort are substantially elevated compared to those reported in other Asian populations. This suggests Saudis may experience more varied drug responses, especially for medications that undergo metabolism by the CYP2C8 enzyme, like ibuprofen., (© 2024 Walter de Gruyter GmbH, Berlin/Boston.)

Energy drink (ED) consumption has become increasingly popular. Due to a lack of evidence, it was crucial to assess the effects of Red Bull (RB) consumption on the rat submandibular salivary gland and the potential therapeutic impact of blueberry (BB). Thirty rats were randomly assigned to five groups. Group 1 (Control) received distilled water. Group 2 (RB) received RB (10 mL/100 g/day) for 8 weeks. Group 3 (BB) rats were administered BB (500 mg/day for 8 weeks). Group 4 (RB + BB (L)) received RB for 8 weeks, and from the 5th week, were concurrently given BB (250 mg/day) for 4 weeks. Group 5 (RB + BB (H)) received RB for 8 weeks, and from the 5th week, were concurrently given BB (500 mg/day) for 4 weeks. At the end of the experiment, blood samples were collected, the animals were euthanized, and their submandibular salivary glands were harvested. Oxidative stress markers (MDA, GPx, CAT, and SOD) were assessed in both serum and tissue. Inflammatory markers (TNF-α, IL-6, and IL-10) were quantified in tissue. Submandibular gland specimens were prepared for light microscopy, and immunohistochemical staining was performed using anti-α-SMA. RB consumption resulted in a significant increase in MDA, TNF-α, IL-6, and IL-10, while GPx, CAT, and SOD levels decreased significantly. Degenerative changes in the gland's structure were observed in the RB group. A significant increase in α-SMA immunoreaction was detected in myoepithelial cells. Administration of BB, particularly at a high dose, ameliorated the aforementioned findings. In conclusion, blueberry administration exhibited therapeutic effects due to its antioxidative and anti-inflammatory properties.

Cisplatin (CP) is a highly effective broad-spectrum chemotherapeutic agent for several solid tumors. However, its clinical use is associated with ovarian toxicity. Icariin (ICA) is a bioactive flavonoid of Epimedium brevicornum with reported protective activities against inflammation, oxidative stress and ovarian failure. This study aimed to explore the protective effects of ICA against CP-associated ovarian toxicity in rats. Rats were randomized into five groups and treated for 17 days: control, ICA (10 mg/kg/day, for 17 days. p.o.), CP (6 mg/kg, i.p. on days 7 and 14), CP + ICA (CP 6 mg/kg i.p. on days 7 and 14 and ICA 5 mg/kg p.o. daily), and CP + ICA (CP 6 mg/kg i.p. on days 7 and 14 and ICA 10 mg/kg p.o. daily). Our results indicated that ICA effectively improved ovarian reserve as indicated by attenuating CP-induced histolopathological changes and enhancing serum anti-müllerian hormone (AMH). Furthermore, co-administration of ICA with CP showed restoration of the oxidant-anti-oxidant balance in ovarian tissues, evidenced by decreased malondialdehyde (MDA) concentrations and elevated superoxide dismutase (SOD) and catalase (CAT) activities. Also, ICA suppressed ovarian inflammation as evidenced by down-regulation of the expression of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and nuclear factor kappa B (NF-κB). ICA inhibited ovarian apoptosis in CP-treated rats by down-regulation of CASP3 and Bax and up-regulation of Bcl-2 mRNA expression. Further, ICA enhanced PTEN, p-AKT, p-mTOR, and p-AMPKα expression. In conclusion, ICA possesses a protective activity against CP-induced ovarian toxicity in rats by exhibiting antioxidant, antiinflammatory, anti-apoptotic activities and modulating NF-κB expression and PTEN/AKT/mTOR/AMPK axis in ovarian tissues., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Bisphenol A (BPA) is commonly used in resin-based dental materials and is categorised as an endocrine-disrupting compound. This study aimed to evaluate the knowledge, attitudes, and practice toward the use of BPA among faculty members and postgraduate students. In this descriptive study, a validated self-administered questionnaire was used, and 204 dentists were included, with a response rate of 56.2%. The findings revealed that faculty members had significantly better knowledge (p=0.024) and practice (p=0.036) and more positive attitudes (p=0.024) toward BPA exposure. Regression analysis showed a significantly positive effect of attending a workshop/lecture or reading an article about BPA on the participants' total mean knowledge (p<0.001) and practice scores (p<0.001). Furthermore, faculty members showed a significantly more positive attitude (p<0.001) toward BPA exposure. These results emphasise the importance of increased awareness and education concerning BPA exposure to ensure optimal dental care.

Chlorpyrifos (CPS), an organophosphorus insecticide, is widely used for agricultural and non-agricultural purposes with hazardous health effects. Berberine (BBR) is a traditional Chinese medicine and a phytochemical with anti-inflammatory and anti-oxidative properties. The present study evaluated the effects of BBR against kidney damage induced by CPS and the underlying mechanisms. An initial study indicated that BBR 50 mg/kg was optimal under our experimental conditions. Then, 24 rats (6/group) were randomized into: control, BBR (50 mg/kg/day), CPS (10 mg/kg/day), and CPS + BBR. BBR was administration 1 h prior to CPS. Each treatment was delivered daily for a period of 28 consecutive days using a gastric gavage tube. Compared to CPS-alone treated rats, BBR effectively improved renal function by preventing the rise in serum urea, creatinine, and uric levels. The reno-protective effects of BBR were confirmed through a histological examination of kidney tissues. BBR restored oxidant-antioxidant balance in renal tissues mediated by Keap1/Nrf2/HO-1 axis modulation. In addition, BBR decreased nitric oxide (NO) and myeloperoxidase (MPO) activity. This was paralleled with the potent down-regulation of NF-κB. Furthermore, BBR exhibited anti-apoptotic activities supported by the upregulation of Bcl-2 and down-regulation of Bax and caspase-3 expression. In conclusion, our data suggest that BBR attenuates CPS-induced nephrotoxicity in rats by restoring oxidant-antioxidant balance and inhibiting inflammatory response and apoptosis in renal tissue. This is mediated, at least partly, by modulation of the Nrf2/HO-1 axis., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Metabolic syndrome (MetS) is a combination of metabolic disorders that can predispose individuals to benign prostatic hyperplasia (BPH). The inhibition of the cannabinoid 1 (CB1) receptor has been used to treat metabolic disorders in animal models. This study reports the use of a peripherally restricted CB1 antagonist (AM6545) and a neutral CB1 antagonist (AM4113) to improve MetS-related BPH in rats. Animals were divided into three control groups to receive either a normal rodent diet, AM6545, or AM4113. MetS was induced in the fourth, fifth, and sixth groups using a concentrated fructose solution and high-salt diet delivered as food pellets for eight weeks. The fifth and sixth groups were further given AM6545 or AM4113 for additional four weeks. Body and prostate weights were measured and prostate sections were stained with hematoxylin eosin. Cyclin D1, markers of oxidative stress and inflammation, and levels of the endocannabinoids were recorded. BPH in rats with MetS was confirmed through increased prostate weight and index, as well as histopathology. Treatment with either AM6545 or AM4113 significantly decreased prostate weight, improved prostate histology, and reduced cyclin D1 expression compared with the MetS group. Groups treated with CB1 antagonists experienced reduced lipid peroxidation, recovered glutathione depletion, restored catalase activity, and had lower inflammatory markers interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α). MetS rats treated with either AM6545 or AM4113 showed reduced concentrations of anandamide (AEA) and 2-arachidonoylglycerol (2-AG) in the prostate compared with the MetS group. In conclusion, the CB1 antagonists AM6545 and AM4113 protect against MetS-induced BPH through their anti-proliferative, antioxidant, and anti-inflammatory effects.

Myocardial injury (MI) is an important pathological driver of mortality worldwide., and arises as a result of imbalances between myocardial oxygen demand and supply. In MI, oxidative stress often leads to inflammatory changes and apoptosis. Current therapies for MI are known to cause various adverse effects. Consequently, the development of new therapeutic agents with a reduced adverse event profile is necessary. In this regard, 2-methoxyestradiol (2ME), the metabolic end-product of oestradiol, possesses anti-inflammatory and antioxidant properties. The aim of this research is to assess the impact of 2ME on cardiac injury caused by isoproterenol (ISO) in rats. Animals were separated into six groups; controls, and those receiving 2ME (1 mg/kg), ISO (85 mg/kg), ISO + 2ME (0.25 mg/kg), ISO + 2ME (0.5 mg/kg), and ISO + 2ME (1 mg/kg). 2ME significantly attenuated ISO-induced changes in electrocardiographic changes and the cardiac histological pattern. This compound also decreased lactate dehydrogenase activity, creatine kinase myocardial band and troponin levels. The ability of 2ME to act as an antioxidant was shown by a decrease in malondialdehyde concentration, and the restoration of glutathione levels and superoxide dismutase activity. Additionally, 2ME antagonized inflammation and cardiac cell apoptosis, a process determined to be mediated, at least partially, by suppression of Gal-3/TLR4/MyD88/NF-κB signaling pathway. 2ME offers protection against acute ISO-induced MI in rats and offers a novel therapeutic management option., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Author(s).)

The single nucleotide polymorphism (SNP) A118G (rs1799971) in the Mu Opioid Receptor 1 (OPRM1) gene is associated with significant variations in analgesic doses and adverse effects of opioids. The A118G OPRM1 allele distributions vary significantly between different populations worldwide. The study aimed to assess the allele frequency and genotype distribution of OPRM1 A118G SNP in Saudis. This cross-sectional study included 124 healthy Saudis (62 males and 62 females) visiting the King Abdulaziz University Hospital in Jeddah, Saudi Arabia. The Oragene®-DISCOVER (OGR-600) kits were used to collect saliva samples from the participants. Polymerase chain reaction-restriction fragment length polymorphism was utilized to assess the SNP. Among the tested population, 79.03% (95% C.I. 70.81-85.82) were homozygous wild-type A118A, 16.13% (95% C.I. 10.14-23.80) were heterozygous A118G, and 4.84% (95% C.I. 1.80-10.23) were homozygous mutant G118G. OPRM1 A118G polymorphism allele frequencies were 87% (95% C.I. 79.89-92.44) and 13% (95% C.I. 7.56-20.11) for the 118A and 118G alleles, respectively. A higher frequency of the OPRM1 118G allele was present in females, 21% (95% C.I. 11.66-33.17) compared to males, 5% (95% C.I. 1.01-13.50). Relative to other Asian countries, the Saudi population showed a low prevalence of the OPRM1 A118G polymorphism, with a higher frequency of the 118G allele in females. Our research will contribute to the existing knowledge on the prevalence of OPRM1 A118G polymorphism, which could be considered for the personalized prescribing of opioid analgesics., Competing Interests: The authors report no conflicts of interest in this work.

This study evaluated the effect of Virtual Reality Distraction (VRD) on dental anxiety among anxious children undergoing prophylactic dental treatment by utilizing both subjective (Venham Anxiety and Behavioral Rating Scale (VABRS)) and objective (heart rate (HR) and salivary cortisol level (SCL)) measures. This randomized controlled study included 36 (6- to 14-year-old) healthy and anxious children who needed prophylactic dental treatment and had a history of previous dental treatment. The eligible children's anxiety level was evaluated using a modified version of the Abeer Dental Anxiety Scale-Arabic version (M-ACDAS) and those who scored at least 14 or more out of 21 were included. Participants were randomly distributed to either the VRD or control group. In the VRD group, participants wore the VRD eyeglasses during prophylactic dental treatment. In the control group, subjects received their treatment while watching a video cartoon on a regular screen. The participants were videotaped during the treatment, and their HR was recorded at four time points. Also, a sample from each participant's saliva was collected twice, at the baseline and after the procedure. The mean M-ACDAS score at baseline in the VRD and the control groups was not statistically significant ( p = 0.424). At the end of the treatment, the SCL was significantly lower in the VRD group ( p < 0.001). Neither the VABRS ( p = 0.171) nor the HR significantly differed between the VRD and control groups. Virtual reality distraction is a non-invasive method that has the potential to significantly reduce anxiety during prophylactic dental treatment among anxious children., Competing Interests: The authors declare no conflict of interest., (©2023 The Author(s). Published by MRE Press.)

Chlorpyrifos (CPF), is an organophosphate pesticide that is widely used for agricultural purposes. However, it has well-documented hepatotoxicity. Lycopene (LCP) is a plant-derived carotenoid with antioxidant and anti-inflammatory activities. The present work was designed to evaluate the potential hepatoprotective actions of LCP against CPF-induced hepatotoxicity in rats. Animals were assigned into five groups namely: Group I (Control), Group II (LCP), Group III (CPF), Group IV (CPF + LCP 5 mg/kg), and Group V (CPF + LCP 10 mg/kg). LCP offered protection as evidenced by inhibiting the rise in serum activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) induced by CPF. This was confirmed histologically as LCP-treated animals showed liver tissues with less proliferation of bile ducts and periductal fibrosis. LCP significantly prevented the rise in hepatic content of malondialdehyde (MDA), depletion of reduced glutathione (GSH), and exhaustion of glutathione-s-transferase (GST) and superoxide dismutase (SOD). Further, LCP significantly prevented hepatocyte death as it ameliorated the increase in Bax and the decrease in Bcl-2 expression induced by CPF in liver tissues as determined immunohistochemically. The observed protective effects of LCP were further confirmed by a significant enhancement in heme oxygenase-1 (HO-1) and NF-E2-related factor 2 (Nrf2) expression. In conclusion, LCP possesses protective effects against CPF-induced hepatotoxicity. These include antioxidation and activation of the Nrf2/HO-1 axis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Nephrotoxicity is a serious complication that limits the clinical use of gentamicin (GEN). Parthenolide (PTL) is a sesquiterpene lactone derived from feverfew with various therapeutic benefits. However, PTL possesses low oral bioavailability. This study aimed to evaluate the therapeutic protective effects of PTL-phytosomes against GEN-induced nephrotoxicity in rats. The PTL was prepared as phytosomes to improve the pharmacological properties with a particle size of 407.4 nm, and surface morphology showed oval particles with multiple edges. Rats were divided into six groups: control, nano-formulation plain vehicle, PTL-phytosomes (10 mg/kg), GEN (100 mg/kg), GEN + PTL-phytosomes (5 mg/kg), and GEN + PTL-phytosomes (10 mg/kg). The administration of PTL-phytosomes alleviated GEN-induced impairment in kidney functions and histopathological damage, and decreased kidney injury molecule-1 (KIM-1). The anti-oxidative effect of PTL-phytosomes was demonstrated by the reduced malondialdehyde (MDA) concentration and increased superoxide dismutase (SOD) and catalase (CAT) activities. Furthermore, PTL-phytosomes treatment significantly enhanced sirtuin 1 (Sirt-1), nuclear factor erythroid-2-related factor-2 (Nrf2), NAD(P)H quinone dehydrogenase 1 (NQO1), and heme oxygenase-1 (HO-1). Additionally, PTL-phytosomes treatment exhibited anti-inflammatory and anti-apoptotic properties in the kidney tissue. These findings suggest that PTL-phytosomes attenuate renal dysfunction and structural damage by reducing oxidative stress, inflammation, and apoptosis in the kidney.

Chemotherapy-induced peripheral neuropathy (CIPN) is one of chemotherapies' most often documented side effects. Patients with CIPN experience spontaneous burning, numbness, tingling, and neuropathic pain in their feet and hands. Currently, there is no effective pharmacological treatment to prevent or treat CIPN. Activating the cannabinoid receptor type 1 (CB 1 ) by orthosteric agonists has shown promising results in alleviating the pain and neuroinflammation associated with CIPN. However, the use of CB 1 orthosteric agonists is linked to undesirable side effects. Unlike the CB 1 orthosteric agonists, CB 1 positive allosteric modulators (PAMs) don't produce any psychoactive effects, tolerance, or dependence. Previous studies have shown that CB 1 PAMs exhibit antinociceptive effects in inflammatory and neuropathic rodent models. This study aimed to investigate the potential benefits of the newly synthesized GAT229, a pure CB 1 PAM, in alleviating neuropathic pain and slowing the progression of CIPN. GAT229 was evaluated in a cisplatin-induced (CIS) mouse model of peripheral neuropathic pain (3 mg/kg/d, 28 d, i.p.). GAT229 attenuated and slowed the progression of thermal hyperalgesia and mechanical allodynia induced by CIS, as evaluated by the hotplate test and von Frey filament test. GAT229 reduced the expression of proinflammatory cytokines in the dorsal root ganglia (DRG) neurons. Furthermore, GAT229 attenuated nerve injuries by normalizing the brain-derived neurotrophic factor and the nerve growth factor mRNA expression levels in the DRG neurons. The CB 1 receptor antagonist/inverse agonist AM251 blocked GAT229-mediated beneficial effects. According to our data, we suggest that CB 1 PAMs might be beneficial in alleviating neuropathic pain and slowing the progression of CIPN., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Author(s).)

Garcinia mangostana L. (Clusiaceae), a popular tropical fruit for its juiciness and sweetness, is an opulent fountain of prenylated and oxygenated xanthones with a vast array of bio-activities. Garcinone E (GE), a xanthone derivative reported from G. mangostana, possesses cytotoxic and aromatase inhibitory activities. The present research endeavors to investigate the hepato-protection efficaciousness of GE on concanavalin-A (Con-A)-instigated hepatitis. Results showed that GE pretreating noticeably diminishes both the serum indices (transaminases, ALP, LDH, and γ-GT) and histopathological lesions of the liver. It counteracted neutrophil and CD4+ infiltration into the liver. GE furthered the Nrf2 genetic expression and its antioxidants' cascade, which resulted in amelioration of Con-A-caused oxidative stress (OS), lipid per-oxidative markers (4-HNE, MDA, PC) reduction, and intensified antioxidants (TAC, SOD, GSH) in the hepatic tissue. Additionally, GE prohibited NF-ĸB (nuclear factor kappa-B) activation and lessened the genetics and levels of downstream cytokines (IL1β and IL6). Moreover, the TNF-α/JNK axis was repressed in GE-treated mice, which was accompanied by attenuation of Con-A-induced apoptosis. These findings demonstrated the protective potential of GE in Con-A-induced hepatitis which may be associated with Nrf2/HO-1 signaling activation and OS suppression, as well as modulation of the NF-κB and TNF-α/JNK/apoptosis signaling pathway. These results suggest the potential use of GE as a novel hepato-protective agent against autoimmune hepatitis.

Objective: The purpose of this research is to assess the commitment of participants in Saudi Arabia and Egypt towards healthy daily habits, preventive measures, healthy food habits, and beliefs about natural products as an immunostimulants during COVID-19 pandemic., Method: A cross-sectional questionnaire-based study was conducted in Saudi Arabia (mainly Riyadh and Jeddah) and Egypt (mainly Cairo). The questionnaire instrument was created based on an extensive literature review on the COVID-19 pandemic, including its spreading and transmission methods, preventive measures, healthy lifestyle, and diets that increase human immunity against viral infections and the use of natural products and drinks. The questionnaire was created by Microsoft 365® office forms, participants were invited through emails and other social media. The questionnaire includes a demographic section (gender, nationality, residency country, city, age, marital status, educational level, employment status, chronic disease history, under anxiety or stress, have a temper or irritable person, were infected/currently infected and in contact to COVID-19 patient) and (23) questions arranged under five domains; Domain I daily habits (4), Domain II keeping preventive measures (4), Domain III healthy eating habits (9), Domain IV for participants currently or previously infected, or in contact with a patient (4) Domain V for assessment of participants' beliefs towards the use of natural products to elevate immunity during COVID-19 pandemic (2), beside 4 choice questions (stimulant drinks, natural drinks, natural products, and zinc-rich food). SPSS® was used to analyze the results using Student' t-test, ANOVA, and Tukey's HSD tests., Result: 510 individuals with various demographic characteristics participated in the study. This study revealed that the participants belief in healthy foods, natural drinks (mainly ginger, lemon, and cinnamon), natural products (mainly honey, olive oil, and black seed), healthy habits, and preventive measures as sanitizers, social distance, and exercise. Only 13% of all participants were infected with COVID-19, although 31% of them were in contact with COVID -19 patients, about 93% were under stress, and 22% were with chronic diseases. Participants who are married, not in contact with patients and not previously infected by COVID-19 are more adhered to preventive measures while those previously or currently infected are more committed to healthy lifestyle and diet habits. Qualification level seems to make no significant difference in any domain. 78.6% of the participants beliefs in the benefits of utilizing natural products in preventing infection with corona virus or reducing the period of treatment in case of infection. About 95.7% of the infected persons had no need of hospitalization and about 50% are cured within two weeks of infection. The questionnaire revealed that Nescafe and black tea were the most used stimulant drinks among the participants, particularly the students and who were always under stress. Most of the participants agreed with the utilization of Zn-rich food, particularly Egyptians, which may help in boosting their immunity., Conclusion: Natural products selected in the present study can be used in combination with the existing clinical standards of care that have the potential to serve as prophylactic agents in populations that are at risk to develop COVID-19 infection., (Copyright: © Pharmacy Practice.)

Endometrial hyperplasia (EH) is the most common risk factor for endometrial malignancy in females. The pathogenesis of EH has been directly linked to uterine inflammation, which can result in abnormal cell division and decreased apoptosis. Piceatannol (PIC), a natural polyphenolic stilbene, is known to exert anti-inflammatory, antioxidant and anti-proliferative activities. The aim of the present study was to examine the potential preventive role of PIC in estradiol benzoate (EB)-induced EH in rats. A self-nanoemulsifying drug delivery system (SNEDDS) was prepared to improve the solubility of the PIC. Therefore, thirty female Wistar rats were divided into five groups: (1) control, (2) PIC SNEDDS (10 mg/kg), (3) EB (0.6 mg/kg), (4) EB + PIC SNEDDS (5 mg/kg) and (5) EB + PIC SNEDDS (10 mg/kg). The administration of PIC SNEDDS prevented EB-induced increases in uterine weights and histopathological changes. Additionally, it displayed pro-apoptotic and antioxidant activity in the endometrium. Immunohistochemical staining of uterine sections co-treated with PIC SNEDDS showed significantly decreased expression of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and nuclear transcription factor-kappa B (NF-κB). This anti-inflammatory effect was further confirmed by a significant increase in Nrf2 and heme oxygenase-1 (HO-1) expression. These results indicate that SNEDDS nanoformulation of PIC possesses protective effects against experimentally induced EH.

Objectives: Cytochrome P450 2E1 ( CYP2E1 ) is one of the major enzymes involved in the metabolism and detoxification of various drugs and xenobiotics. Polymorphisms in the CYP2E1 gene exhibit high inter-individual variations associated with alterations in CYP2E1 gene expression and enzyme function. This study aimed to determine the genotype distributions and allele frequencies of CYP2E1*1B, *5B , and *6 polymorphisms among Saudis in western Saudi Arabia., Methods: In total, 140 healthy Saudis attending King Abdulaziz University Hospital between February and April 2021 were included in the study. CYP2E1 gene polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism analysis., Results: The genotype frequencies of CYP2E1*1B A2A2, A2A1, and A1A1 were 54.29%, 40%, and 8%, respectively. The frequencies of CYP2E1*5B c1c1 and c1c2 genotypes were approximately 99.93% and 0.07%, respectively. The frequencies of the CYP2E1*6 DD, DC, and CC genotypes were 91.43%, 7.85%, and 0.72%, respectively. The genotype distributions for these polymorphisms were consistent with the expected distribution based on Hardy-Weinberg equilibrium. The allele frequencies were 74.29% A2 and 25.71% A1 for CYP2E1*1B , 99.64% c1 and 0.36% c2 for CYP2E1*5B , and 95.36% D and 4.65% C for CYP2E1*6 ., Conclusion: The genotype distribution of CYP2E1*1B polymorphism was higher in the western Saudi population, whereas the CYP2E1*5B and *6 polymorphisms were lower than the global average. Knowledge of the prevalence of CYP2E1 polymorphisms among our population will provide a better understanding of whether individual patients might benefit from their medication or whether they might develop adverse effects., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Author(s).)

Objectives: This study compared and evaluated the clinical and radiographic efficacy of non-instrumentation triple antibiotic paste pulp therapy and Vitapex pulpectomy in non-vital primary molars., Material and Methods: Healthy, 5-9 years old children with at least one non-vital primary molar were included in the study. Molars were divided into two groups based on the subject's cooperation level. In the first group, molars received triple antibiotic paste, and a second group received Vitapex pulpectomy followed by a stainless-steel crown. Triple antibiotic paste was freshly prepared and proportioned in equal parts by volume (metronidazole, minocycline, and ciprofloxacin = 1:1:1) before the scheduled treatment. A clinical and radiographic examination was performed by two trained and calibrated pediatric dentists at the pre-operative baseline and the 6- and 12-month follow-up visits., Results: A total of 28 molars received triple antibiotic paste pulp therapy and 20 received Vitapex pulpectomy. At the 6-month follow-up, the success rate among the molars in the triple antibiotic paste group was clinically (92.85%) and radiographically (85.71%) higher compared to the Vitapex group (91.67%, 62.50% respectively) with p = 0.89 and 0.55 respectively. At the 12-month follow-up, the molars in the triple antibiotic paste group showed lower clinical (95.45%) but higher radiographic success rate (72.73%) compared to the Vitapex group (100% and 62.50%) with (p = 0.85 and 0.47) respectively. None of the differences were statistically significant., Conclusions: Both triple antibiotic paste and Vitapex can be clinically and radiographically effective in treating non-vital primary molars., (© 2021 The Authors. Clinical and Experimental Dental Research published by John Wiley & Sons Ltd.)

The human brain requires adequate cerebral blood flow to meet the high demand for nutrients and to clear waste products. With age, there is a chronic reduction in cerebral blood flow in small resistance arteries that can eventually limit proper brain function. The endothelin system is a key mediator in the regulation of cerebral blood flow, but the contributions of its constituent receptors in the endothelial and vascular smooth muscle layers of cerebral arteries have not been well defined in the context of aging. We isolated posterior cerebral arteries from young and aged Fischer 344 rats, as well as ET B receptor knock-out rats and mounted the vessels in plexiglass pressure myograph chambers to measure myogenic tone in response to increasing pressure and targeted pharmacological treatments. We used an ET A receptor antagonist (BQ-123), an ET B receptor antagonist (BQ-788), endothelin-1, an endothelin-1 synthesis inhibitor (phosphoramidon), and vessel denudation to dissect the roles of each receptor in aging vasculature. Aged rats exhibited a higher myogenic tone than young rats, and the tone was sensitive to the ET A antagonist, BQ-123, but insensitive to the ET B antagonist, BQ-788. By contrast, the tone in the vessels from young rats was raised by BQ-788 but unaffected by BQ-123. When the endothelial layer that is normally enriched with ET B1 receptors was removed from young vessels, myogenic tone increased. However, denudation of the endothelial layer did not influence vessels from aged animals. This indicated that endothelial ET B1 receptors were not functional in the vessels from aged rats. There was also an increase in ET A receptor expression with age, whereas ET B receptor expression remained constant between young and aged animals. These results demonstrate that in young vessels, ET B1 receptors maintain a lower myogenic tone, but in aged vessels, a loss of ET B receptor activity allows ET A receptors in vascular smooth muscle cells to raise myogenic tone. Our findings have potentially important clinical implications for treatments to improve cerebral perfusion in older adults with diseases characterized by reduced cerebral blood flow.

Diabetic peripheral neuropathy (DPN) is characterized by progressive loss of peripheral nerves, which causes numbness, weakness, and severe pain. The medications available currently provide only modest relief from the pain of DPN and are associated with various side effects, which has generated an enormous demand for research on new therapeutic approaches. Dysregulation of the endocannabinoid system has been reported in DPN. Cannabinoid-based medications have gained increasing attention as a potential therapy to alleviate DPN pain. Endocannabinoids and cannabinoids' actions are mediated primarily by cannabinoid receptor 1 (CB 1 R) and cannabinoid receptor 2 (CB 2 R). Cannabinoids that activate CB 1 R have demonstrated a profound antinociceptive effect, although CB 1 R is associated with undesirable psychoactive effects. Peripherally restricted CB 1 R agonists help overcome this problem; however, adverse metabolic and cardiovascular effects limit its therapeutic use. In contrast, CB 1 R antagonists, selective CB 2 R agonists, and endocannabinoid metabolizing enzymes inhibitors alleviate DPN pain effectively with minimal side effects. This article provides a concise overview of the preclinical and clinical studies that have tested the therapeutic potential of targeting the endocannabinoid system to treat painful DPN., Competing Interests: There are no conflicts of interest., (Copyright: © 2021 Journal of Microscopy and Ultrastructure.)

Background: Methylenetetrahydrofolate reductase, the encoded by the MTHFR gene, plays a crucial role in converting the amino acid homocysteine to methionine. Two polymorphisms of the MTHFR gene, C677T and A1298C, reportedly reduce enzyme activity, resulting in hyperhomocysteinemia. Patients with C677T and A1298C polymorphisms may be at higher risk for developing abnormal hyperhomocysteinemia, which has been linked to catastrophic neurological including fatal outcomes., Objective: Determine the prevalence of the MTHFR gene variants C677T and A1298C among pediatric dental patients treated at King Abdulaziz University Hospital., Design: Cross-sectional., Setting: Clinics of pediatric dentistry department., Subjects and Methods: Healthy Saudi children 6-12 years old with no known allergies were screened for eligibility between May and December 2019. A single investigator collected saliva samples. The MTHFR C677T and A1298C polymorphisms were analyzed using polymerase chain reaction and restriction fragment length polymorphism., Main Outcome Measure: The prevalence of MTHFR gene variants (C677T and A1298C) among the subjects., Sample Size: 138., Results: MTHFR C677T polymorphism was present in 36.2% of the sample and 90.0% of children carrying this allele were heterozygotes. MTHFR A1298C polymorphism was present in 91.3% of the sample and 77.0% of the children carrying this allele were heterozygotes. No linkage disequilibrium between MTHFR C677T and MTHFR A1298C was observed within this sample., Conclusions: Our study found a high frequency of the MTHFR A1298C genotype, which was substantially more abundant than expected based on a Hardy-Weinberg distribution. Therefore, caution is advised in using N 2 O in Saudi children as the increased prevalence of this MTHFR allele may increase the incidence of serious adverse effects among these children., Limitations: Further studies are recommended with a larger sample size from randomly selected hospitals from different regions of Saudi Arabia., Conflict of Interest: None.

Background: Integrating pharmacogenetics (PGx) testing into clinical practice leads to personalized medicine, which improves treatments' efficacy and safety. Successful implementation of such a service requires sufficient knowledge, perception, and self-confidence among healthcare providers, especially pharmacists., Objectives: To evaluate governmental hospital pharmacists' knowledge, perception, and self-confidence toward PGx testing in Jeddah, Kingdom of Saudi Arabia., Method: This cross-sectional study was conducted using previously validated questionnaire. Pharmacists working in five randomly selected general governmental hospitals in Jeddah between August and October 2019 were interviewed. Comparative and descriptive analyses were used to analyze the data, and the significance level was at P -value < 0.05., Results: A total of 119 pharmacists with a mean (±SD) age of 31.2 (±5.05) years were included with a response rate of 79.3%. The average total mean (±SD) score for PGx knowledge-based questions was low (2.4 ± 1.09 out of 5). Most of the participants, with a total mean score of (10.1 ± 1.6 out of 12), revealed a positive perception toward PGx testing and its implications. A moderate self-confidence score for utilizing PGx testing (4.3 ± 2.3 out of 8) was observed among the participants. Pharmacists who had completed postgraduate studies had a statistically higher mean knowledge score ( P  = 0.006) compared with pharmacists with undergraduate degrees., Conclusion: Governmental hospital pharmacists have limited knowledge and understanding about PGx testing; however, the majority expressed a high level of awareness and agreed that PGx testing is a valuable tool for enhancing drug efficacy and safety. The study also highlighted the importance of improving pharmacists' knowledge about PGx testing, which will help them in implementing such a valuable service into their clinical practice in Saudi hospitals., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2020 The Author(s).)

The cannabinoid type 1 (CB 1 ) receptor and the dopamine type 2 (D 2 ) receptor are co-localized on medium spiny neuron terminals in the globus pallidus where they modulate neural circuits involved in voluntary movement. Physical interactions between the two receptors have been shown to alter receptor signaling in cell culture. The objectives of the current study were to identify the presence of CB 1 /D 2 heteromers in the globus pallidus of C57BL/6J male mice, define how CB 1 /D 2 heteromer levels are altered following treatment with cannabinoids and/or antipsychotics, and determine if fluctuating levels of CB 1 /D 2 heteromers have functional consequences. Using in situ proximity ligation assays, we observed CB 1 /D 2 heteromers in the globus pallidus of C57BL/6J mice. The abundance of the heteromers increased following treatment with the nonselective cannabinoid receptor agonist, CP55,940. In contrast, treatment with the typical antipsychotic haloperidol reduced the number of CB 1 /D 2 heteromers, whereas the atypical antipsychotic olanzapine treatment had no effect. Co-treatment with CP55,940 and haloperidol had similar effects to haloperidol alone, whereas co-treatment with CP55,940 and olanzapine had similar effects to CP55,940. The observed changes were found to have functional consequences as the differential effects of haloperidol and olanzapine also applied to γ-aminobutyric acid release in STHdh Q7/Q7 cells and motor function in C57BL/6J male mice. This work highlights the clinical relevance of co-exposure to cannabinoids and different antipsychotics over acute and prolonged time periods., (© 2020 Wiley Periodicals LLC.)

The endothelin receptor A (ETA) and endothelin receptor B (ETB) are G protein-coupled receptors that are co-expressed in vascular smooth muscle cells. Endothelin-1 (ET-1) activates endothelin receptors to cause microvascular vasoconstriction. Previous studies have shown that heteromerization between ETA and ETB prolongs Ca 2+ transients, leading to prolongation of Gα q -dependent signaling and sustained vasoconstriction. We hypothesized that these effects are in part mediated by the resistance of ETA/ETB heteromers to β-arrestin recruitment and subsequent desensitization. Using bioluminescence resonance energy transfer 2 (BRET 2 ), we found that ETB has a relatively equal affinity to form either homomers or heteromers with ETA when co-expressed in the human embryonic kidney 293 (HEK293) cells. When co-expressed, activation of ETA and ETB by ET-1 caused a heteromer-specific reduction and delay in β-arrestin-2 recruitment with a corresponding reduction and delay in ET-1-induced ETA/ETB co-internalization. Furthermore, the co-expression of ETA and ETB inhibited ET-1-induced β-arrestin-1-dependent extracellular signal-regulated kinase (ERK) phosphorylation while prolonging ET-1-induced Gα q -dependent ERK phosphorylation. ETA/ETB heteromerization mediates the long-lasting vasoconstrictor response to ET-1 by the prolongation of Gα q -dependent signaling and inhibition of β-arrestin function.

Background: Attention-deficit/hyperactivity disorder (ADHD) and lower cognitive ability have been linked with increased likelihood of exposure to adversity. We hypothesized that these associations may be partly due to genetic factors., Methods: We calculated polygenic scores for ADHD and intelligence and assessed psychopathology and general cognitive ability in a sample of 297 youth aged 5-27 years enriched for offspring of parents with mood and psychotic disorders. We calculated an adversity score as a mean of 10 indicators, including socio-economic disadvantage, childhood maltreatment and bullying. We tested the effects of polygenic scores, externalizing symptoms and IQ on adversity scores using mixed-effects linear regression., Results: Externalizing symptoms and general cognitive ability showed expected positive and negative relationships with adversity, respectively. Polygenic scores for intelligence were unrelated to adversity, but polygenic scores for ADHD were associated with adversity (β = 0.23, 95% CI 0.13 to 0.34, p < .0001). ADHD polygenic scores uniquely explained 4.0% of variance in adversity score. The relationship between polygenic scores for ADHD and adversity was independently significant among individuals with (β = 0.49, 95% CI 0.25 to 0.75, p < .0001) and without (β = 0.14, 95% CI 0.02 to 0.26, p = .022) ADHD., Conclusions: A genetic score indexing liability to ADHD was associated with exposure to adversity in early life. Previously observed associations between externalizing symptoms, lower cognitive ability and adversity may be partially attributed to genetic liability to ADHD., (© 2019 Association for Child and Adolescent Mental Health.)

Huntington's disease (HD) is an inherited progressive neurodegenerative disease characterized by motor, cognitive, and behavioural changes. One of the earliest changes to occur in HD is a reduction in cannabinoid 1 receptor (CB 1 ) levels in the striatum, which is strongly correlated with HD pathogenesis. CB 1 positive allosteric modulators (PAM) enhance receptor affinity for, and efficacy of activation by, orthosteric ligands, including the endocannabinoids anandamide and 2-arachidonoylglycerol. The goal of this study was to determine whether the recently characterized CB 1 allosteric modulators GAT211 (racemic), GAT228 (R-enantiomer), and GAT229 (S-enantiomer), affected the signs and symptoms of HD. GAT211, GAT228, and GAT229 were evaluated in normal and HD cell models, and in a transgenic mouse model of HD (7-week-old male R6/2 mice, 10 mg/kg/d, 21 d, i.p.). GAT229 was a CB 1 PAM that improved cell viability in HD cells and improved motor coordination, delayed symptom onset, and normalized gene expression in R6/2 HD mice. GAT228 was an allosteric agonist that did not enhance endocannabinoid signaling or change symptom progression in R6/2 mice. GAT211 displayed intermediate effects between its enantiomers. The compounds used here are not drugs, but probe compounds used to determine the potential utility of CB 1 PAMs in HD. Changes in gene expression, and not protein, were quantified in R6/2 HD mice because HD pathogenesis is associated with dysregulation of mRNA levels. The data presented here provide the first proof of principle for the use of CB 1 PAMs to treat the signs and symptoms of HD., (Copyright © 2019. Published by Elsevier Ltd.)

G protein-coupled receptors (GPCRs) are the target for many drugs. Evidence continues to accumulate demonstrating that multiple receptors form homo- and heteromeric complexes, which in turn dynamically couple with G proteins, and other interacting proteins. Here, we describe a method to simultaneously determine the identity of up to four distinct constituents of GPCR complexes using a combination of sequential bioluminescence resonance energy transfer 2-fluorescence resonance energy transfer (SRET 2 ) with bimolecular fluorescence complementation (BiFC). The method is amenable to moderate throughput screening of changes in response to ligands and time-course analysis of protein-protein oligomerization.

G protein-coupled receptors (GPCRs) interact with multiple intracellular effector proteins such that different ligands may preferentially activate one signal pathway over others, a phenomenon known as signaling bias. Signaling bias can be quantified to optimize drug selection for preclinical research. Here, we describe moderate-throughput methods to quantify signaling bias of known and novel compounds. In the example provided, we describe a method to define cannabinoid-signaling bias in a cell culture model of Huntington's disease (HD). Decreasing type 1 cannabinoid receptor (CB 1 ) levels is correlated with chorea and cognitive deficits in HD. There is evidence that elevating CB 1 levels and/or signaling may be beneficial for HD patients while decreasing CB 1 levels and/or signaling may be detrimental. Recent studies have found that Gα i/o -biased CB 1 agonists activate extracellular signal-regulated kinase (ERK), increase CB 1 protein levels, and improve viability of cells expressing mutant huntingtin. In contrast, CB 1 agonists that are β-arrestin1-biased were found to reduce CB 1 protein levels and cell viability. Measuring agonist bias of known and novel CB 1 agonists will provide important data that predict CB 1 -specific agonists that might be beneficial in animal models of HD and, following animal testing, in HD patients. This method can also be applied to study signaling bias for other GPCRs.

Type 1 cannabinoid (CB 1 ) and dopamine 2 long form (D 2L ) receptors can physically interact to form heteromers that display unique pharmacology in vitro compared to homomeric complexes. Co-expression of CB 1 and D 2L and co-application of CB 1 and D 2 agonists increases cAMP levels while administration of either agonist alone decreases cAMP levels. To understand the observed co-agonist response, our first goal of the current study was to define the stoichiometry of CB 1 /D 2L /Gα protein complexes. Using bioluminescence resonance energy transfer 2 (BRET 2 ), we confirmed that, CB 1 homodimers, D 2L homodimers, and CB 1 /D 2L heteromers are formed. By using sequential resonance energy transfer 2 (SRET 2 ) combined with bimolecular fluorescence complementation (BiFC), we were able to demonstrate that CB 1 /D 2L form heterotetramers consisting of CB 1 and D 2L homodimers. We demonstrated that CB 1 /D 2L heterotetramers are coupled to at least two Gα proteins. The second aim of the study was to investigate allosteric effects of a D 2L agonist (quinpirole) on CB 1 receptor function and to investigate the effects of a CB 1 agonist [arachidonyl-2-chloroethylamide (ACEA)] on D 2L receptor function within CB 1 /D 2L heterotetramers. Treating cells co-expressing CB 1 and D 2L with both ACEA and quinpirole switched CB 1 and D 2L receptor coupling and signaling from Gα i to Gα s proteins, enhanced β-arrestin1 recruitment and receptor co-internalization. The concept of bidirectional allosteric interaction within CB 1 /D 2 heterotetramers has important implications for understanding the activity of receptor complexes in native tissues and under pathological conditions., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The G protein-coupled cannabinoid receptors CB 1 , CB 2 , GPR18, and GPR55 regulate neurotransmission, pain, and inflammation and have been intensively investigated as potential drug targets. Each of these GPCRs is coupled to multiple effector proteins mediating divergent cellular signals. The ligand bias of cannabinoid-targeted compounds is only beginning to be quantified. Research into cannabinoid bias is now revealing correlations between bias in cell culture and functional outcomes in vivo. We present an example study of cannabinoid bias in the context of Huntington disease. In future, an understanding of cannabinoid receptor structure and quantification of ligand bias will optimize drug selection matched to patient population and disease., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Activation of dopamine receptor 2 long (D2L) switches the signaling of type 1 cannabinoid receptor (CB1) from Gαi to Gαs, a process thought to be mediated through CB1-D2L heteromerization. Given the clinical importance of D2 antagonists, the goal of this study was to determine if D2 antagonists could modulate CB1 signaling. Interactions between CB1 and D2L, Gαi, Gαs, and β-arrestin1 were studied using bioluminescence resonance energy transfer 2 (BRET(2)) in STHdh(Q7/Q7) cells. CB1-dependent extracellular regulated kinase (ERK)1/2, CREB phosphorylation, and CB1 internalization following cotreatment of CB1 agonist and D2 antagonist were quantified. Preassembled CB1-Gαi complexes were detected by BRET(2) Arachidonyl-2'-chloroethylamide (ACEA), a selective CB1 agonist, caused a rapid and transient increase in BRET efficiency (BRETEff) between Gαi-Rluc and CB1-green fluorescent protein 2 (GFP(2)), and a Gαi-dependent increase in ERK phosphorylation. Physical interactions between CB1 and D2L were observed using BRET(2) Cotreatment of STHdh(Q7/Q7) cells with ACEA and haloperidol, a D2 antagonist, inhibited BRETEff signals between Gαi-Rluc and CB1-GFP(2) and reduced the EMax and pEC50 of ACEA-mediated Gαi-dependent ERK phosphorylation. ACEA and haloperidol cotreatments produced a delayed and sustained increase in BRETEff between Gαs-Rluc and CB1-GFP(2) and increased the EMax and pEC50 of ACEA-induced Gαs-dependent cAMP response element-binding protein phosphorylation. In cells expressing CB1 and D2L treated with ACEA, binding of haloperidol to D2 receptors switched CB1 coupling from Gαi to Gαs In addition, haloperidol treatment reduced ACEA-induced β-arrestin1 recruitment to CB1 and CB1 internalization. D2 antagonists allosterically modulate cannabinoid-induced CB1 coupling, signaling, and β-arrestin1 recruitment through binding to CB1-D2L heteromers. These findings indicate that D2 antagonism, like D2 agonists, change agonist-mediated CB1 coupling and signaling., (Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics.)

Huntington disease (HD) is an inherited, autosomal dominant, neurodegenerative disorder with limited treatment options. Prior to motor symptom onset or neuronal cell loss in HD, levels of the type 1 cannabinoid receptor (CB1) decrease in the basal ganglia. Decreasing CB1 levels are strongly correlated with chorea and cognitive deficit. CB1 agonists are functionally selective (biased) for divergent signaling pathways. In this study, six cannabinoids were tested for signaling bias in in vitro models of medium spiny projection neurons expressing wild-type (STHdh(Q7/Q7)) or mutant huntingtin protein (STHdh(Q111/Q111)). Signaling bias was assessed using the Black and Leff operational model. Relative activity [ΔlogR (τ/KA)] and system bias (ΔΔlogR) were calculated relative to the reference compound WIN55,212-2 for Gαi/o, Gαs, Gαq, Gβγ, and β-arrestin1 signaling following treatment with 2-arachidonoylglycerol (2-AG), anandamide (AEA), CP55,940, Δ(9)-tetrahydrocannabinol (THC), cannabidiol (CBD), and THC+CBD (1:1), and compared between wild-type and HD cells. The Emax of Gαi/o-dependent extracellular signal-regulated kinase (ERK) signaling was 50% lower in HD cells compared with wild-type cells. 2-AG and AEA displayed Gαi/o/Gβγ bias and normalized CB1 protein levels and improved cell viability, whereas CP55,940 and THC displayed β-arrestin1 bias and reduced CB1 protein levels and cell viability in HD cells. CBD was not a CB1 agonist but inhibited THC-dependent signaling (THC+CBD). Therefore, enhancing Gαi/o-biased endocannabinoid signaling may be therapeutically beneficial in HD. In contrast, cannabinoids that are β-arrestin-biased--such as THC found at high levels in modern varieties of marijuana--may be detrimental to CB1 signaling, particularly in HD where CB1 levels are already reduced., (Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics.)

Background and Purpose: Cannabidiol has been reported to act as an antagonist at cannabinoid CB1 receptors. We hypothesized that cannabidiol would inhibit cannabinoid agonist activity through negative allosteric modulation of CB1 receptors., Experimental Approach: Internalization of CB1 receptors, arrestin2 recruitment, and PLCβ3 and ERK1/2 phosphorylation, were quantified in HEK 293A cells heterologously expressing CB1 receptors and in the STHdh(Q7/Q7) cell model of striatal neurons endogenously expressing CB1 receptors. Cells were treated with 2-arachidonylglycerol or Δ(9)-tetrahydrocannabinol alone and in combination with different concentrations of cannabidiol., Key Results: Cannabidiol reduced the efficacy and potency of 2-arachidonylglycerol and Δ(9)-tetrahydrocannabinol on PLCβ3- and ERK1/2-dependent signalling in cells heterologously (HEK 293A) or endogenously (STHdh(Q7/Q7)) expressing CB1 receptors. By reducing arrestin2 recruitment to CB1 receptors, cannabidiol treatment prevented internalization of these receptors. The allosteric activity of cannabidiol depended upon polar residues being present at positions 98 and 107 in the extracellular amino terminus of the CB1 receptor., Conclusions and Implications: Cannabidiol behaved as a non-competitive negative allosteric modulator of CB1 receptors. Allosteric modulation, in conjunction with effects not mediated by CB1 receptors, may explain the in vivo effects of cannabidiol. Allosteric modulators of CB1 receptors have the potential to treat CNS and peripheral disorders while avoiding the adverse effects associated with orthosteric agonism or antagonism of these receptors., (© 2015 The British Pharmacological Society.)

The endocannabinoid system (ECS) and the dopaminergic system (DAS) are two major regulators of basal ganglia function. During Huntington's disease (HD) pathogenesis, the expression of genes in both the ECS and DAS is dysregulated. The purpose of this study was to determine the changes that were consistently observed in the ECS and DAS during HD progression in the central nervous system (CNS) and in the periphery in different models of HD and human HD tissue. To do this, we conducted a meta-analysis of differential gene expression in the ECS and DAS using publicly available microarray data. The consolidated data were summarized as observed changes in gene expression (OCGE) using a weighted sum for each gene. In addition, consolidated data were compared to previously published studies that were not available in the gene expression omnibus (GEO) database. The resulting data confirm gene expression changes observed using different approaches and provide novel insights into the consistency between changes observed in human tissue and various models, as well as disease stage- and tissue-specific transcriptional dysregulation in HD. The major implication of the systems-wide data presented here is that therapeutic strategies targeting the ECS or DAS must consider the dynamic changes in gene expression over time and in different body areas, which occur during HD progression and the interconnectedness of the two systems.

Modulation of type 1 cannabinoid receptor (CB1) activity has been touted as a potential means of treating addiction, anxiety, depression, and neurodegeneration. Different agonists of CB1 are known to evoke varied responses in vivo. Functional selectivity is the ligand-specific activation of certain signal transduction pathways at a receptor that can signal through multiple pathways. To understand cannabinoid-specific functional selectivity, different groups have examined the effect of individual cannabinoids on various signaling pathways in heterologous expression systems. In the current study, we compared the functional selectivity of six cannabinoids, including two endocannabinoids (2-arachidonyl glycerol (2-AG) and anandamide (AEA)), two synthetic cannabinoids (WIN55,212-2 and CP55,940), and two phytocannabinoids (cannabidiol (CBD) and Δ(9)-tetrahydrocannabinol (THC)) on arrestin2-, Gα(i/o)-, Gβγ-, Gα(s)-, and Gα(q)-mediated intracellular signaling in the mouse STHdh(Q7/Q7) cell culture model of striatal medium spiny projection neurons that endogenously express CB1. In this system, 2-AG, THC, and CP55,940 were more potent mediators of arrestin2 recruitment than other cannabinoids tested. 2-AG, AEA, and WIN55,212-2, enhanced Gα(i/o) and Gβγ signaling, with 2-AG and AEA treatment leading to increased total CB1 levels. 2-AG, AEA, THC, and WIN55,212-2 also activated Gα(q)-dependent pathways. CP55,940 and CBD both signaled through Gα(s). CP55,940, but not CBD, activated downstream Gα(s) pathways via CB1 targets. THC and CP55,940 promoted CB1 internalization and decreased CB1 protein levels over an 18-h period. These data demonstrate that individual cannabinoids display functional selectivity at CB1 leading to activation of distinct signaling pathways. To effectively match cannabinoids with therapeutic goals, these compounds must be screened for their signaling bias., (© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.)

The human type 1 cannabinoid (hCB1) receptor is expressed at high levels in the central nervous system. mRNA variants of the coding region of this receptor, human cannabinoid hCB1a and hCB1b receptors, have been identified, their biological function remains unclear. The present study demonstrated that the three human cannabinoid hCB1 coding region variants are expressed in the human and monkey (Macaca fascicularis) brain. Western blot analyses of homogenates from different regions of the monkey brain demonstrated that proteins with the expected molecular weights of the cannabinoid CB1, CB1a and CB1b receptors were co-expressed throughout the brain. Given the co-localization of these receptors, we hypothesized that physical interactions between the three splice variants may affect cannabinoid pharmacology. The human cannabinoid hCB1, hCB1a, and hCB1b receptors formed homodimers and heterodimers, as determined by BRET in transiently transfected HEK 293A cells. We found that the co-expression of the human cannabinoid hCB1 and each of the splice variants increased cell surface expression of the human cannabinoid hCB1 receptor and increased Gi/o-dependent ERK phosphorylation in response to cannabinoid agonists. Therefore, the human cannabinoid hCB1 coding region splice variants play an important physiological role in the activity of the endocannabinoid system., (© 2013 Elsevier B.V. All rights reserved.)