Highlights on the Development, Related Patents, and Prospects of Lenacapavir: The First-in-Class HIV-1 Capsid Inhibitor for the Treatment of Multi-Drug-Resistant HIV-1 Infection.

The multidrug-resistant (MDR) human immunodeficiency virus 1 (HIV-1) infection is an unmet medical need. HIV-1 capsid plays an important role at different stages of the HIV-1 replication cycle and is an attractive drug target for developing therapies against MDR HIV-1 infection. Lenacapavir (LEN) is the first-in-class HIV-1 capsid inhibitor approved by the USFDA, EMA, and Health Canada for treating MDR HIV-1 infection. This article highlights the development, pharmaceutical aspects, clinical studies, patent literature, and future directions on LEN-based therapies. The literature for this review was collected from PubMed, authentic websites (USFDA, EMA, Health Canada, Gilead, and NIH), and the free patent database (Espacenet, USPTO, and Patent scope). LEN has been developed by Gilead and is marketed as Sunlenca (tablet and subcutaneous injection). The long-acting and patient-compliant LEN demonstrated a low level of drug-related mutations, is active against MDR HIV-1 infection, and does not reveal cross-resistance to other anti-HIV drugs. LEN is also an excellent drug for patients having difficult or limited access to healthcare facilities. The literature has established additive/synergistic effects of combining LEN with rilpivirine, cabotegravir, islatravir, bictegravir, and tenofovir. HIV-1 infection may be accompanied by opportunistic infections such as tuberculosis (TB). The associated diseases make HIV treatment complex and warrant drug interaction studies (drug-drug, drug-food, and drug-disease interaction). Many inventions on different aspects of LEN have been claimed in patent literature. However, there is a great scope for developing more inventions related to the drug combination of LEN with anti-HIV/anti-TB drugs in a single dosage form, new formulations, and methods of treating HIV and TB co-infection. Additional research may provide more LEN-based treatments with favorable pharmacokinetic parameters for MDR HIV-1 infections and associated opportunistic infections such as TB.

Evaluating the effect of gargling with hydrogen peroxide and povidone-iodine on salivary viral load of SARS-CoV-2: A pilot randomized clinical trial.

Background and Aim: This study evaluates the salivary viral load of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in hospitalized patients and outpatients before and after gargling with 1% hydrogen peroxide and 0.25% povidone-iodine in comparison with normal saline. Patients and Methods: This clinical trial was conducted on 120 participants with laboratory-confirmed coronavirus disease 2019 (COVID-19) in two groups: outpatients (n = 60) and hospitalized patients (n = 60). In each group, the patients were randomly divided into three subgroups of 20 based on their given mouthwash for gargling (hydrogen peroxide, povidone-iodine, or normal saline). Two saliva samples were taken from each patient: the first one before gargling and the second one 10 minutes after gargling 10 ml of the respected mouthwashes for 30 seconds. The TaqMan real-time polymerase chain reaction (PCR) amplification of SARS-CoV-2 was used to measure the viral load. Results: Saliva samples from 46% of patients were positive for coronavirus before gargling the mouthwashes. The percentage of patients with an initial positive saliva sample was significantly higher in the outpatient group (83.3%) than in the hospitalized group (5.4%) (P = 0.01). According to the findings, gargling any mouthwash similar to saline did not reduce the viral load (P > 0.05). Conclusion: The saliva of COVID-19 patients in the initial stage of the disease was more likely to contain SARS-CoV-2 than the saliva of the hospitalized patients. Gargling hydrogen peroxide or povidone-iodine did not reduce the salivary SARS-CoV-2 viral load.

Regulation of mitochondrial temperature in health and disease.

Mitochondrial temperature is produced by various metabolic processes inside the mitochondria, particularly oxidative phosphorylation. It was recently reported that mitochondria could normally operate at high temperatures that can reach 50℃. The aim of this review is to identify mitochondrial temperature differences between normal cells and cancer cells. Herein, we discussed the different types of mitochondrial thermosensors and their advantages and disadvantages. We reviewed the studies assessing the mitochondrial temperature in cancer cells and normal cells. We shed the light on the factors involved in maintaining the mitochondrial temperature of normal cells compared to cancer cells.

Use of a novel 1-hour protocol for rapid frozen section immunocytochemistry, in a case of squamous cell carcinoma treated with Mohs micrographic surgery.

For squamous cell carcinoma (SCC) treated using Mohs micrographic surgery (MMS), interpretation of haematoxylin and eosin-stained frozen sections can be challenging. In these situations, ancillary use of immunostaining is a useful tool for the Mohs surgeon. However, use of immunostaining in MMS laboratories is limited, mainly because current manual immunostaining platforms are subject to operator error, and automated immunostaining, albeit accurate, is too slow for inclusion in MMS. In this report, we describe a novel 1-hour protocol for rapid frozen section immunocytochemistry, using the pancytokeratin markers AE1/AE3. This protocol has been specifically designed to integrate the speed of manual techniques and the accuracy of automated platforms, making it a valuable addition to the MMS laboratory. We propose that in selected or histologically challenging cases, there is a role for the use of this novel protocol, allowing the Mohs surgeon to more confidently declare tumour clearance, thus preventing further unnecessary surgery and preserving healthy tissue.

New embedding and staining systems PrestoCHILL and Presto stainer for application in the advancement of Mohs micrographic surgery.

BACKGROUND: Mohs micrographic surgery (MMS) involves evaluation of frozen tissue sections to determine complete circumferential and deep tissue margin clearance of skin tumours. PrestoCHILL and Presto stainer devices are two new innovative tools which bring benefits of automation, speed and efficiency to the preparation of frozen section analysis in MMS. The devices were assessed at Viapath's Tissue Science Mohs laboratory at Guy's Cancer Centre. MATERIAL AND METHODS: A total of 279 samples from 10 anatomically different facial sites. These included nose (95), lip (24), forehead (47), cheek (25), eyelids (34), temple (9), chin (15), ear (17), scalp (6) and neck (7). These were analysed using both devices simultaneously. RESULTS: The PrestoCHILL device was measured for accuracy of tissue orientation by determining how many of the cases examined microscopically had complete margin and full epidermis preservation. The precision and reproducibility of the Presto stainer was evaluated by the consistency of achieving ideal standards of staining quality as defined by the department's internal quality control check, on stained sections examined and evaluated microscopically. The mean (standard deviation) score for accuracy for the PrestoCHILL across all tissue facial sites was 93.5 (11)%; the mean (standard deviation) score for precision/reproducibility of the Presto stainer was 96.5 (11)% (both p < 0.05). CONCLUSION: The devices combined offer an assured accuracy and precision performance, which is reproducible across all facial tissue types examined. The devices represent a key step forward in the introduction of improved automated embedding and staining procedures within MMS.

A multicentre study of the precision and accuracy of the TruSlice and TruSlice Digital histological dissection devices.

BACKGROUND: Five key factors enabling a good surgical grossing technique include a flat uniformly perpendicular specimen cutting face, appropriate immobilisation of the tissue specimen during grossing, good visualisation of the cutting tissue face, sharp cutting knives and the grossing knife action. TruSlice and TruSlice Digital are new innovative tools based on a guillotine configuration. The TruSlice has plastic inserts whilst the TruSlice Digital has an electronic micrometre attached: both features enable these dissection factors to be controlled. The devices were assessed in five hospitals in the UK. MATERIAL AND METHODS: A total of 267 fixed tissue samples from 23 tissue types were analysed, principally the breast (n = 32) skin (30), rectum (28), colon (27) and cervix (17). Precision and accuracy were evaluated by measuring the defined thickness, and the consistency of achieving the defined thickness of tissue samples taken respectively. Both parameters were expressed as a total percentage of compliance for the cohort of samples accessed. RESULTS: Overall, the mean (standard deviation) score for precision was 81 (11) % whilst the accuracy score was 82 (11) % (both p < 0.05, chi-squared test), although this varied with type of tissue. Accuracy and precision were strongly correlated (rp = 0.83, p < 0.001). CONCLUSION: The TruSlice Digital devices offer an assured precision and accuracy performance which is reproducible across an assortment of tissue types. The use of a micrometre to set tissue slice thickness is innovative and should comply with laboratory accreditation requirements, alleviating concerns of how to tackle issues such as the 'measurement of uncertainty' at the grossing bench.

Retrospective pilot study for analysis of antidepressant serum concentrations of citalopram and venlafaxine during inflammation.

INTRODUCTION: Inflammation-mediated changes in drug metabolism may increase drug levels in blood and lead to intoxications. The objective of this study was to find out whether elevated serum levels of C-reactive protein (CRP) are associated with increased serum concentrations of the antidepressants citalopram and venlafaxine. METHODS: Therapeutic drug monitoring request forms of psychiatric patients were screened retrospectively. The serum concentrations in relation to the daily doses [(C/D) (ng/ml/mg)] and the metabolic ratios (metabolite/drug) were compared intraindividually under normal (<5 mg/l) and pathological (>5 mg/l) condition by the Wilcoxon signed-rank test. RESULTS: Elevated levels of CRP were not associated with a significant (P>0.05) increase in C/D for citalopram (2.4 ng/ml/mg vs. 2.85 ng/ml/mg, N=15) or in C/D for the active moiety of venlafaxine (1.76 ng/ml/mg vs. 1.68 ng/ml/mg, N=39), compared with normal CRP serum levels. No significant difference in the metabolic ratio was observed in both groups. DISCUSSION: There was no major effect of inflammation on the metabolism of citalopram and venlafaxine. Because of the broad therapeutic indices of these 2 drugs, the drugs seem to be a good choice for the treatment of depression, even if an infection occurs.

Pan-cytokeratin markers for rapid frozen section immunocytochemistry from head and facial Mohs cases of basal cell carcinoma: a comparison and evaluation to determine the marker of choice.

The application of immunocytochemistry in the field of Mohs micrographic surgery (MMS) is well established. This study evaluates the use of pan-cytokeratins (AE1/AE3, MNF116 and AE1/AE3+PCK26) in the assessment of basal cell carcinoma (BCC) on frozen tissue debulk specimens. Fifty-five cases of BCC, all from head and facial sites, were assessed in the study. In addition to staining all cases for the three cytokeratin antibodies under investigation, sections were also stained with haematoxylin and eosin (H&E) to demonstrate tumour architecture and morphology. All sections for immunocytochemistry were stained on a Roche Ventana BenchMark Ultra automated platform employing a rapid frozen section protocol. Results were assessed based on the intensity of staining of keratinocytes (scale: 0-100%), as well as sensitivity of staining determined by the total percentage of keratinocytes stained within the tissue section. AE1/AE3 demonstrated the most consistent staining both in terms of intensity of staining and sensitivity, with a mean of 99.1% and 99.9%, respectively. AE1/AE3+PCK26 average results indicated scores of 70.6% for intensity and 87.2% for sensitivity, with MNF116 scoring 92.9% for intensity but only 57.3% for sensitivity. The data indicate that AE1/AE3 is the best pan-cytokeratin antibody to use in the assessment of BCC in MMS. The use of cytokeratin immunocytochemistry is justified in morphologically complex cases of BCC, or in cases where dense inflammatory infiltrate surrounding any suspicious cells make identification of small numbers of tumour cells difficult to determine with just an H&E stain. The significant rationale is that cytokeratin staining is a valuable adjunct in the study of tumour cell assessment in cases of MMS for BCC. In addition, the use of anti-AE1/AE3 cytokeratin antibodies provides the most consistent staining results for such cases.

Development of new and accurate measurement devices (TruSlice and TruSlice Digital) for use in histological dissection: an attempt to improve specimen dissection precision.

Histological dissection of human tissue has relied on conventional procedures, which have largely remained unchanged for decades. Practices to determine measurement parameters employed in these procedures have largely relied on the use of rulers and weighing scales. It is well documented in the scientific literature that both fixation and processing of tissue can significantly affect the viability of the of tissue sections both for tinctorial and immunocytochemical investigations. Both of these factors can be compounded in their negative effects by inappropriate sampling of tissue at histological cut up. There are five key factors to ensure good surgical grossing technique, flat uniformly perpendicular specimen cutting face, appropriate immobilisation of the tissue specimen during grossing, good visualisation of the cutting tissue face, sharp cutting knives and the grossing knife action. Meeting these factors implies the devices are fit for purpose. Here we describe an innovative approach to designing cut up devices to improve accuracy and precision, which take these five key requirements into consideration. The devices showed accuracy and precision, enabling tissue slices to be produced in a uniformly perpendicular fashion to within 2 mm in thickness and to enable consistency and reproducibility of performance across a series of tissue types. The application of a digital rule on one of these devices ensures accuracy and also enables quality control issues to be clearly assessed. As cellular pathology laboratories conform to ever increasing standards of compliance and performance in practice, the advent of assured precision and accuracy at cut up is awaited. Recommendations from accreditation bodies such as the United Kingdom Accreditation Service (UKAS) continue to push for improvements in this area of histological investigation. These newly designed devices may give the answers to these requirements and provide the impetus for a new generation of innovative equipment for histological dissection.

Molecular Investigation of Brucella Species Belongs to Sheep and Goats in Seropositive Samples from an Endemic Area of Hamedan Province; Famenin Brucellosis Cohort Study.

Brucellosis is a zoonotic infectious disease in western regions of Iran, especially in Hamedan province. Following the Famenin brucellosis cohort study, the main aim of the current study was the molecular detection of Brucella species (spp.) in sheep and goats from Famenin, Hamedan, Iran. A total of 23 Brucella-seropositive samples (sheep=21 and goats=2), which had been screened from 1,660 animals in the Famenin cohort study, were used to detect Brucella-DNA using the BCSP31 target gene and IS711 locus. In total, 20 of 23 samples were positive for Brucella infection by using specific primers. Additionally, Brucella melitensis (B. melitensis) and Brucella abortus (B. abortus) were confirmed in 90% (n=18) and 10% (n=2) of positive samples, respectively. There was no sample with the co-infection of B. abortus and B. melitensis. In this study, B. abortus was isolated from one of the goat samples. This is the first report on Brucella spp. in animals in the region. It was found that B. melitensis is the dominant spp. responsible for brucellosis in animals from Famenin. Molecular techniques are reliable tools to detect Brucella infection, especially in cases without serology findings and conclusive results.

Transgender Youth's Perspectives on Factors Influencing Intended and Unintended Pregnancies.

Drawing on data from focus groups with 152 trans youth aged 14-18 years in the United States, this article explores the factors that the participants understood as contributing to adolescent pregnancy among trans youth. Youth posited that unintended pregnancies occur due to barriers to contraceptives; a lack of gender-affirming sexual health education; sexual assault and dating violence; and mental health-influenced sexual risk-taking. Participants suggested that intended pregnancies may be a self-development strategy; a self-directed effort to repress/change gender modality or identity; and due to the perceived incompatibility between pregnancy and transition, where pregnancy must occur prior to transitioning.

A Systematic Review on the Relationship Between Physical Activity and Sexual Function in Adults.

Sexual function is a vital component of overall well-being and quality of life. Physical activity is known to have a profound influence on various aspects of health, but its impact on sexual function in the general population remains an under-explored area. This systematic review seeks to thoroughly examine existing evidence, aiming to establish the correlation between physical activity and sexual function in both male and female adults. We conducted a comprehensive search of electronic databases and relevant sources, following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Eligible studies were those that investigated the effects of physical activity on sexual function using the International Index of Erectile Dysfunction (IIEF-5) questionnaire and the Female Sexual Function Index (FSFI). Quality assessment was performed on the included studies, and the findings were synthesized through qualitative analysis. The review identified 12 randomized controlled trials, primarily focusing on males, with interventions ranging from home-based walking to structured exercise training. Only two studies were conducted among females. The most recommended exercise was aerobic exercise. Consistent aerobic exercise proves to be a hopeful and efficient non-drug intervention for enhancing erectile function in men. However, when considering the effects of physical exercise programs on sexual function and the quality of sexual life of females, the results present challenges in drawing clear conclusions. Health policymakers play an important role in providing guidelines and recommendations to healthcare professionals, encouraging them to prescribe exercise as a preferable alternative to pharmacological treatments for enhancing sexual functions in both men and women.

Dermatofibrosarcoma protuberans: dealing with slow Mohs procedures employing formalin-fixed, paraffin wax-embedded tissue in a busy diagnostic laboratory.

Dermatofibrosarcoma protuberans (DFSP) is a relatively uncommon tumour that arises in the dermis and underlying soft tissue. Surgical removal is the preferred treatment, with relatively wide clearance margins of 3 cm or more. Slow Mohs procedures are often employed successfully to treat patients with such tumours. Slow Mohs procedures offer the benefit of improved cure rates and maximal tissue conservation. However, dealing with such tissue successfully presents the laboratory with a host of technical problems. This report advocates a set protocol to follow for slow Mohs, based on the experience acquired from dealing with 37 cases of DFSP over a 12-year period. The report establishes the benefits of slow Mohs paraffin wax-embedded tissue over frozen sections in terms of improved morphology, tissue preservation and immunocytochemical labelling with anti-CD34.

Personality, density and habitat drive the dispersal of invasive crayfish.

There is increasing evidence that personality traits may drive dispersal patterns of animals, including invasive species. We investigated, using the widespread signal crayfish Pacifastacus leniusculus as a model invasive species, whether effects of personality traits on dispersal were independent of, or affected by, other factors including population density, habitat, crayfish size, sex and limb loss, along an invasion gradient. Behavioural traits (boldness, activity, exploration, willingness to climb) of 310 individually marked signal crayfish were measured at fully-established, newly-established and invasion front sites of two upland streams. After a period at liberty, recaptured crayfish were reassessed for behavioural traits (newly-established, invasion front). Dispersal distance and direction of crayfish movement, local population density, fine-scale habitat characteristics and crayfish size, sex and limb loss were also measured. Individual crayfish exhibited consistency in behavioural traits over time which formed a behavioural syndrome. Dispersal was both positively and negatively affected by personality traits, positively by local population density and negatively by refuge availability. No effect of size, sex and limb loss was recorded. Personality played a role in promoting dispersal but population density and local habitat complexity were also important determinants. Predicting biological invasion in animals is likely to require better integration of these processes.

Fish community and abundance response to improved connectivity and more natural hydromorphology in a post-industrial subcatchment.

Barrier removal and fish pass construction are increasingly used as tools to restore river connectivity and improve habitat quality, but the effectiveness of subcatchment-scale connectivity restoration on recovery of fish communities is poorly understood. We used a before-after-downstream-upstream methodology to determine the effects of subcatchment-scale connectivity restoration on fishes in a fragmented tributary of the River Wear, Northeast England, between 2013 and 2019. Following restoration (three barriers removed, five barriers fitted with fish passes, two barriers unaltered), riffle habitat increased, fine sediment decreased, and most fish species benefitted. Total fish abundance, comprising seven native species, increased 3 years after the restoration and remained elevated to the end of the study. Mean brown trout (Salmo trutta) density increased from 20.9 ±â€¯6.3 to 33.8 ±â€¯16.8 per 100m2 from 2013 to 2019, with Young-of-Year trout increasing from 10.6 ±â€¯4.6 to 19.8 ±â€¯11.8 per 100m2. Connectivity restoration reduced the mean age of trout, suggesting a change to an increased migratory component of the population. Density of bullhead (Cottus perifretum), a species with poor dispersal ability, increased from 4.6 ±â€¯2.7 to 32.6 ±â€¯17.9 per 100m2 over 2013 to 2019. Stone loach (Barbatula barbatula), also a less mobile species but tolerant to fine sediment, decreased in abundance where barriers were removed. Atlantic salmon (Salmo salar) were absent over the study timescale, despite being common in the Wear, and despite suitable habitat and water quality in the restored subcatchment, suggesting a hysteresis effect. Our findings indicate that, where good water quality exists, restoring river connectivity and hydromorphology at a subcatchment scale is beneficial for most native resident and migratory fishes. However, the ecological benefits of connectivity restoration, especially in rivers with many barriers, may take several years to develop. We encourage well-controlled long-term studies reporting the outcomes of large-scale connectivity restoration.

Evaluation of different haematoxylin stain subtypes for the optimal microscopic interpretation of cutaneous malignancy in Mohs frozen section histological procedure.

BACKGROUND: The Mohs technique employs mainly H&E-stained frozen sections for surgical margin assessment of cutaneous excisions, utilising microscopic evaluation of the complete, circumferential, peripheral and deep margins. This study aimed to determine which mordant based haematoxylin (Ehrlich's, Cole's, Mayer's, Gill's I, Gill's II, Gill's III, Weigert's, Harris' or Carazzi's) produced the optimal morphological clarity of staining for the identification of cellular and tissue morphology of cutaneous basal cell carcinoma (BCC). MATERIAL AND METHODS: In total, 100 anonymised patient cases were selected, sectioned and stained with each haematoxylin subtype. The slides were independently evaluated microscopically by two assessors. A combined score was generated to determine the sensitivity (defined as the intensity of haematoxylin staining being too weak or too strong and the colour appearance of the haematoxylin not being blue/black) and specificity (defined as the appearance of background staining with haematoxylin, uneven staining and staining deposits) for each of the nine haematoxylin subtypes. The scoring criteria were based on the UKNEQAS CPT Mohs procedure assessment criteria. RESULTS: The scores generated for specificity identified Carazzi's haematoxylin as best performing (99.2%) followed by Gill's III (98.4%), Ehrlich's (98.2%) and Harris' (85.0%). The sensitivity score again identified Carazzi's as producing the best result (85.0%) followed by Weigert's (83.4%), Ehrlich's (81.6%) and Gill's III (80.4%). DISCUSSION: Carazzi's haematoxylin is the most optimal staining dye for the identification of BCC tumour for use as part of the Mohs micrographic surgery procedure.

Pharmacodynamics of a losartan transdermal system for the treatment of hypertension.

AIMS: Transdermal therapeutic systems were developed using the polymers, Eudragit E 100 and polyvinyl pyrrolidone VA 64 in a film casting assembly. The medicated films were evaluated for physical properties, in vitro drug release studies, in vitro skin permeation studies, and pharmacodynamic studies. RESULTS: The physical parameters were found to be very satisfactory with high drug content (>99%). The in vitro drug release studies were performed using paddle-over-disc assembly specified in USP XXIII. The pharmacodynamic studies were carried out using tail cuff method in Wistar albino rats. Hypertension was induced by methyl prednisolone acetate subcutaneously for 2 weeks. The developed matrix patch was found to decrease the blood pressure (25.42% reduction in mean systolic blood pressure of rats) significantly (P < 0.001) in proximity of the normal value and it was maintained for 24 hours. CONCLUSION: It can be concluded that the developed transdermal matrix patch holds promise for the management of hypertension that needs to be validated by clinical trials.

Pharmacokinetics of a losartan potassium released from a transdermal therapeutic system for the treatment of hypertension.

Monolithic transdermal therapeutic systems (TTS) were developed for sustained antihypertensive effect of losartan potassium using the polymers Eudragit E 100 and polyvinyl pyrrolidone VA 64. The developed formulations (polymeric films) were evaluated for physical characteristics, ex vivo (histopathology) and in vivo (pharmacokinetic studies). Pharmacokinetic parameters, such as C(max), t(max), and AUC were estimated. The transdermal formulation in the present study was found to enhance the relative bioavailability of losartan potassium by 2.2 times with reference to an oral delivery. The increased bioavailability might be due to elimination of hepatic first pass metabolism. Thus, the transdermal formulation F3E with polymeric composition of Eudragit E 100 and polyvinyl pyrrolidone VA 64 (5:3) was found to provide prolonged steady state concentrations of losartan potassium with minimal fluctuations and improved bioavailability.