Content and Activity of Cytochrome P450 3A in Rat Brain Microsomes and Mitochondria

The potential contribution of CYP3A in brain microsomes and mitochondria is crucial to designing appropriate therapeutic strategies and preventing toxicities. Until recently, there has been limited data reported in the literature with respect to CYP3A activity and its expression in the brain tissue. The reason for this limitation is most likely the relatively low level of CYP3A in the brain tissue. To this end, the primary purpose of this study is to develop a CYP3A functional assay in rat brain mitochondria and microsomes to determine the Michaelis-Menten kinetics of the formation of CYP3A-mediated hydroxylated metabolites of midazolam by UPLC-MS/MS. We also aim to analyze the protein levels of CYP3A in the rat brain mitochondria and microsomes by Western blot analysis. Microsome and mitochondria were prepared using different centrifugation methods, followed by purification of the crude mitochondrial fractions using the Percoll density gradient method. The purity of the fractions was determined by Western blot analysis of the microsomal (calreticulin) and mitochondrial (VDAC) markers. Different concentrations of midazolam were incubated with rat brain microsomes and mitochondria in a side-by-side comparison over a period of 10 minutes. The method was developed and optimized to determine the Michaelis-Menten kinetics of MDZ 1′- and 4-hydroxylase activities in rat brain microsomes and mitochondria (n=8). For 4- hydroxymidazolam, brain mitochondria had 2.5-fold higher maximum velocity and 5-fold higher MM constant compared to microsomes. For 1’-hydroxymidazolam, mitochondria had 9-fold higher maximum velocity and 153-fold higher MM constant compared to microsomes. Consistent with the higher Vmax of the metabolites in the mitochondrial fractions, the Western blot analysis showed higher band intensities related to CYP3A2 in the brain mitochondria, when compared with the brain microsomes. Our in vitro study indicates that in the brain the content and activity of CYP3A in the mitochondria are higher than those in the microsomes, an observation which is opposite of the pattern reported in the liver. The local metabolism of CYP3A substrates in the brain microsomes and mitochondria may have significant ramifications for the effectiveness or toxicity of the centrally-acting drugs that are CYP3A substrates

Differential Expression and Activities of Cytochrome P450 3A in the Rat Brain Microsomes and Mitochondria

Midazolam (MDZ), a benzodiazepine derivative, is metabolized to 1′- and 4-hydroxylated metabolites (1′-OH-MDZ and 4-OH-MDZ, respectively) by cytochrome P450 3A (CYP3A). The purpose of this study was to investigate the CYP3A-mediated hydroxylation of MDZ in the rat brain mitochondria (MT). Brain microsomes (MC) and MT fractions were prepared from rats (n = 8) using differential and density gradient centrifugations, and the purity of the fractions was evaluated using VDAC1 and calreticulin as markers of MT and MC, respectively. The formation rates of 1′-OH-MDZ and 4-OH-MDZ in the rat brain MC and MT samples were determined using an LC–MS/MS method after validation. Subsequently, Michaelis–Menten kinetics of 1′- and 4-hydroxylation of MDZ were estimated. Western blot (WB) analysis was used to determine the protein expression of CYP3A in the rat brain MC and MT. The MC fractions had 5.93% ± 3.01% mitochondrial impurity, and the MT fractions had 19.3% ± 7.8% microsomal impurity (mean ± SD). The maximum velocity (Vmax) values of the formation of the hydroxylated metabolites in the brain MT were 2.4–9-fold higher than those in MC. Further, the Vmax values of 4-OH-MDZ in both MC and MT fractions were substantially higher than those of 1′-OH-MDZ. The WB analysis showed that the intensity of the CYP3A immunoreactive band in MT was more than twofold higher than that in MC. It is concluded that compared with MC, rat brain MT contains substantial CYP3A, which may affect the pharmacology or toxicology of centrally acting xenobiotic and endogenous substrates of this enzyme

Squint Among Adult Population in Hail City, Saudi Arabia

Background: Strabismus, also known as crossed eyes, is a condition in which the eyes do not properly align with each other when looking at an object. If present during a large part of childhood, it may result in amblyopia. If onset is during adulthood, it is more likely to result in double vision. Objective: The aim of the study was to estimate the prevalence of squint, types and treatment characteristics in the studied adults in Hail city, Saudi Arabia. Methods: A cross-sectional study conducted in Hail city, Saudi Arabia. The study included 294 participants; 95 male and 199 female adults aged >20 years. The study period was from 1 January to 30 April 2018. Data collected by personal interview using a pre-designed questionnaire, which distributed among the participants to be self-reported. Results: The prevalence of squint among the studied population was 9.9%. it was more common in females than males. Squint was right sided in 24.1% of the cases, left sided in 51.7% and in both eyes in 24.1% of the studied cases. About half (44.8%) of cases had inward squint (esotropia) and 10.3% outward squint (exsotropia), 24.1% of the cases had Intermittent squint and 6.9% had permanent squint. Most (55.2%) of squint cases use glasses and 34.5% of cases squint affected their visual acuity. In 6.9%, squint causes psychological troubles. As regards treatment, 20.7% received medical treatment and 17.2% received surgical treatment but 62.1% do not seek medical care. Only 10.3% of cases completely cured and 13.8% had recurrence. There was insignificant relation with age, sex, education, squint in parents, chronic diseases, consanguinity or hereditary diseases (P>0.05). Conclusion: in this study, the prevalence of squint in the adult participants in Hail city, Saudi Arabia was 9.9% but 62.1% do not seek medical care. After treatment, only 10.3% of cases completely cured and 13.8% had recurrence. Health education of the public about importance of early treatment is highly recommended. Keywords: Squint; strabismus; adult population; prevalence; types; Hail; Saudi Arabia

Examining bedtime procrastination, study engagement, and studyholism in undergraduate students, and their association with insomnia

IntroductionCompulsive overstudying, known as studyholism, is an emerging behavioral addiction. In this study, we examine the prevalence of, and the relationships between, insomnia, study engagement, studyholism, bedtime procrastination among undergraduate students.MethodsThe Studyholism (SI-10), Athens Insomnia (AIS), and bedtime procrastination scales were administered to a convenience sample of 495 university students.ResultsOur findings indicate that the prevalence of insomnia was 75.31%, high studyholism was found in 15.31% of the sample, and increased study engagement was detected in 16.94%. Gender differences analysis revealed that females reported higher studyholism and bedtime procrastination than males. Fifth-year students had higher levels of studyholism than internship (p < 0.001), first-year (p < 0.01), and sixth-year students (p < 0.05). Insomnia was positively related to studyholism and bedtime procrastination. Furthermore, insomnia can be positively predicted by studyholism and bedtime procrastination. Participants with a medium level of studyholism were twice as likely to experience insomnia as those with a low level. Studyholics were six times more susceptible to insomnia than students with low studyholism levels. Compared to individuals with low bedtime procrastination levels, those with medium and high bedtime procrastination were twice as likely to report insomnia.ConclusionOur study highlights the interplay between insomnia, studyholism, and bedtime procrastination. Further, the findings indicate the need to increase awareness of insomnia

Comparison of Generalized Anxiety and Sleep Disturbance among Frontline and Second-Line Healthcare Workers during the COVID-19 Pandemic

Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) infection, also known as COVID-19, has developed into an alarming situation around the world. Healthcare workers are playing the role of frontline defense to safeguard the lives of everyone during the COVID-19 pandemic. The present study aimed to investigate the anxiety levels and sleep quality among frontline and second-line healthcare workers during the COVID-19 pandemic. In this cross-sectional study, a validated, self-administered, electronic questionnaire was distributed through email to healthcare workers. The selection of 1678 healthcare workers was based on a convenience sampling technique. The General Anxiety Disorder-7 (GAD-7) and Pittsburgh Sleep Quality Index (PSQI) instrument scales were used to assess healthcare workers’ anxiety levels and sleep quality during the COVID-19 pandemic. Out of 1678 respondents, 1200 (71.5%) were frontline healthcare workers, while 478 (28.5%) were second-line healthcare workers. Among all the healthcare workers, 435 (25.92%) were experiencing moderate to severe anxiety. Among them, 713 (59.4%) frontline healthcare workers were experiencing anxiety in comparison with 277 (57.9%) second-line healthcare workers. Severe anxiety symptoms were seen in 137 (11.41%) frontline healthcare workers compared to 44 (9.20%) second-line healthcare workers. In total, 1376 (82.0%) healthcare workers were found to have poor sleep quality; 975 (58.10%) were frontline, and 407 (23.89%) were second-line healthcare workers. The highest poor sleep quality levels were found among 642 (84.6%) of the healthcare workers who work in frontline areas (emergency departments, intensive care units, and wards) compared to 734 (79.9%) of the healthcare workers who work in second-line areas. These findings provide a substantial contribution to the consolidation of evidence concerning the negative impact of the pandemic on the mental health of healthcare workers (HCWs). These results have established an association that the COVID-19 pandemic causes larger negative psychological symptoms in frontline healthcare workers, such as severe anxiety and poor sleep quality. Preventive measures to minimize anxiety levels and maintain sleep quality, addressing this issue nationally and globally, are essential to support the healthcare workers who are sacrificing their mental health for the future of our nations

Pharmacological Modulation of Toll-Like Receptors in Brain Disorders

The knowledge regarding pathological and treatment resistance mechanisms involved in the pathology of complex brain disorders is far from understood. The neuroinflammation hypothesis of psychiatric, neurological, and neurodegenerative diseases is well-acknowledged. However, this hypothesis is far from understood. Toll-like receptors (TLRs) family is an innate immunity molecule implicated in neuroinflammation in complex brain disorders. This chapter reviews considerable evidence indicating that activation of endotoxins such as lipopolysaccharide is a common factor. Additionally, we report clinical and preclinical studies highlighting the link between lipopolysaccharide, TLRs, and different types of brain disorders. Also, we review the current pharmacological modulations of TLRs. Hoping we would help in filling our knowledge gaps and highlight potential links to tackle new angles in managing complex brain disorders. This chapter’s primary goal is to encourage scientists and researchers to conduct future studies characterizing the nature of endotoxin activation of TLRs in complex brain disorders, filling our knowledge gaps, and finding new treatment strategies

G Protein-Coupled Receptor Regulation in Cardiovascular Disease: Role of G Protein-Coupled Receptor Kinases

G protein-coupled receptor kinases (GRKs), the negative regulators of G protein-coupled receptors (GPCRs), have a key role in cardiovascular disease pathophysiology. Alteration in GRKs’ expressions and/or kinase activity has been reported in preclinical animal models as well as in patients with cardiovascular diseases. This alteration might be a contributing factor to disease progression by a variety of mechanisms such as non-canonical transduction pathways. The current chapter is aimed to expand our knowledge and understanding of the function of GRKs in cardiovascular diseases, highlight their involvement, and illustrate the possible mechanistic role of GRKs in hypertensive vascular diseases and cardiac myopathy. The current chapter also is endeavoured to identify the potential molecular mechanisms by which GRKs participate in cardiovascular disease progression. Building the basics knowledge about GRKs in cardiovascular diseases will help to assess the potential utilization of GRKs as therapeutic targets and to examine the possible approaches to modulate their protein expression or to inhibit their kinase activity to prevent or attenuate cardiovascular disease progression

The Impact of COVID-19 Crisis on the Control and Management of Radiography Practice in the United Arab Emirates

The present study aimed to assess the impact of the COVID-19 crisis on radiology practices in Abu Dhabi, UAE. An electronic survey (Google form) was distributed among Abu Dhabi government and private hospitals. The survey included general X–ray services, which were only provided in the radiology departments. The diagnostic radiographers who reported changes in the number and type of radiology procedures (37%) reported that the changes reached 61–80% compared to the number of procedures being conducted prior to the outbreak of COVID-19. While infection control was challenging due to the shortage of personal protective equipment (PPE), 51.2% of the participants were affected. The healthcare workers in the radiology departments in Abu Dhabi are exposed to a high number of COVID-19–infection patients, which increases their chances of contracting the disease. A total of 90% of employees were infected with COVID-19 during the crisis. COVID-19 has resulted in changes in clinical working patterns, such as the type and number of procedures performed daily. Additionally, PPE shortages, staff infection during the pandemic, an increase in workplace–related difficulties, and staff well–being are common consequences of the pandemic. It is vital to enhance coping strategies in order to support staff well–being. However, the psychological effects caused as a result of the pandemic should not be ignored, and providing professional support to workers is recommended

Examining Anxiety, Sleep Quality, and Physical Activity as Predictors of Depression among University Students from Saudi Arabia during the Second Wave of the COVID-19 Pandemic

Conducted during the second wave of the pandemic, this cross-sectional study examined the link between sleep quality, physical activity, exposure, and the impact of COVID-19 as predictors of mental health in Saudi undergraduate students. A convenience sample of 207 participants were recruited, 89% of whom were females and 94% were single. The measures included questionnaires on the level of exposure and the perceived impact of COVID-19, a physical activity measure, GAD-7, PHQ-9, and PSQI. The results indicated that approximately 43% of participants exhibited moderate anxiety, and 50% were at risk of depression. Overall, 63.93% of students exposed to strict quarantine for at least 14 days (n = 39) exhibited a high risk of developing depression (χ2(1) = 6.49, p < 0.05, ϕ = 0.18). A higher risk of depression was also found in students whose loved ones lost their jobs (χ2(1) = 4.24, p < 0.05, ϕ = 0.14). Moreover, there was also a strong association between depression and anxiety (β = 0.33, p < 0.01), sleep quality (β = 0.32, p < 0.01), and the perceived negative impact of COVID-19 on socio-economic status (β = 0.26, p < 0.05), explaining 66.67% of depression variance. Our study highlights the socio-economic impact of this pandemic and the overwhelming prevalence of depression

Perspective Chapter: Ketamine, Depression, and Gender Bias

Our knowledge regarding pathological and treatment resistance mechanisms involved in depression is far from understood. Sexual dimorphism in this topic is well acknowledged. However, the need to highlight sex-based discrepancies is unmet. Ketamine, the dissociative anesthetic, has emerged as a rapid antidepressant. This chapter reviewed sexual dimorphism in pharmacological and genetic models of depression, emphasizing ketamine-related antidepressant effects. Aiming by this report, we would extend our knowledge, highlight gender as one of the vital factors in examining depression in preclinical studies, and elucidate complex antidepressant effects associated with ketamine administration. Our central goal is to encourage neuroscientists to consider gender in their studies of mood disorders