Computationally prioritized drugs inhibit SARS-CoV-2 infection and syncytia formation.

The pharmacological arsenal against the COVID-19 pandemic is largely based on generic anti-inflammatory strategies or poorly scalable solutions. Moreover, as the ongoing vaccination campaign is rolling slower than wished, affordable and effective therapeutics are needed. To this end, there is increasing attention toward computational methods for drug repositioning and de novo drug design. Here, multiple data-driven computational approaches are systematically integrated to perform a virtual screening and prioritize candidate drugs for the treatment of COVID-19. From the list of prioritized drugs, a subset of representative candidates to test in human cells is selected. Two compounds, 7-hydroxystaurosporine and bafetinib, show synergistic antiviral effects in vitro and strongly inhibit viral-induced syncytia formation. Moreover, since existing drug repositioning methods provide limited usable information for de novo drug design, the relevant chemical substructures of the identified drugs are extracted to provide a chemical vocabulary that may help to design new effective drugs.

Analysis of aristolochlic acids and evaluation of antibacterial activity of Aristolochia clematitis L.

INTRODUCTION: Several Aristolochia species were used as medicinal herb across Europe and in recent years, their antimicrobial activity has also been investigated. MATERIALS AND METHODS: In this study, A. clematitis was selected to evaluate the aristolochic acids I and II (AA I and AA II) concentrations and the antimicrobial activity of methanol, hexane, butanol, and ethyl acetate extracts of the root, stem, leaf, root, and fruit. AA I and AA II contents were measured by a validated high-performance liquid chromatography-ultraviolet method. RESULTS: Each fraction of the plant contained AA I and AA II and the root was found to have the highest contents of AA I (1.09%) and AA II (0.7454%). The minimum inhibitory concentrations of all extracts were determined by standard microdilution method. The fruit's extracts showed the most efficient antimicrobial effect against both methicillin sensitive and resistant Staphylococcus aureus strains. CONCLUSION: Correlation between the AA I and AA II concentrations and the antimicrobial effect was not found.

ß-Alanine supplementation increased physical performance and improved executive function following endurance exercise in middle aged individuals.

BACKGROUND: Sarcopenia, a reduction in muscle mass and function seen in aging populations, may be countered by improving systemic carnosine stores via beta-Alanine (ß-alanine) supplementation. Increasing systemic carnosine levels may result in enhanced anti-oxidant, neuro-protective and pH buffering capabilities. This enhancement should result in improved exercise capacity and executive function. METHODS: Twelve healthy adults (average age = 60.5 ± 8.6 yrs, weight = 81.5 ± 12.6 kg) were randomized and given either 2.4 g/d of ß-alanine (BA) or Placebo (PL) for 28 days. Exercise capacity was tested via bouts on a cycle ergometer at 70% VO2 peak. Executive function was measured by Stroop Tests 5 min before exercise (T1), immediately before exercise (T2), immediately following fatigue (T3), and 5 min after fatigue (T4). Lactate measures were taken pre/post exercise. Heart rate, Rating of Perceived Exertion (RPE) and VO2 were recorded throughout exercise testing. RESULTS: PRE average time-to-exhaustion (TTE) for the PL and BA group were not significantly different (Mean ± SD; 9.4 ± 1.4mins vs 11.1 ± 2.4mins, respectively, P = 0.7). POST BA supplemented subjects cycled significantly longer than PRE (14.6 ± 3.8mins vs 11.1 ± 2.4mins, respectively, P = 0.04) while those given PL did not (8.7 ± 2.4mins vs 9.4 ± 1.4mins, respectively, P = 0.7). PL subjects were slower in completing the Stroop test POST at T4 compared to T3 (T3 = - 13.3 ± 8.6% vs T4 = 2.1 ± 8.3%, P = 0.04), while the BA group (T3 = - 9.2 ± 6.4% vs T4 = - 2.5 ± 3.5%, P = 0.5) was not. POST lactate production expressed a trend when comparing treatments, as the BA group produced 2.4 ± 2.6 mmol/L more lactate than the PL group (P = 0.06). Within group lactate production for BA (P = 0.4) and PL (P = 0.5), RPE (P = 0.9) and heart rate (P = 0.7) did not differ with supplementation. CONCLUSION: BA supplementation increased exercise capacity and eliminated endurance exercise induced declines in executive function seen after recovery. Increased POST TTE coupled with similar PRE vs POST lactate production indicates an improvement in the ability of BA to extend exercise durations. Furthermore, by countering endurance exercise's accompanying deficits in executive function, the aging population can maintain benefits from exercise with improved safety.

Chemodiversity of Cirsium fruits: Antiproliferative lignans, neolignans and sesquineolignans as chemotaxonomic markers.

While analyzing the fruit composition of nine European Cirsium species representing three sections (i.e., Cephalonoplos, Chamaeleon and Eriolepis), four lignans, three neolignans and three sesquineolignans were determined and used as chemotaxonomic markers. Among them, desmethyl balanophonin and desmethyl picrasmalignan were determined for the first time in the plant kingdom, as the main metabolites of the Chamaeleon section. Prebalanophonin and prepicrasmalignan, identified so far exclusively in C. eriophorum, were also confirmed in C. boujartii and C. vulgare, highlighting the chemotaxonomic significance of these compounds in the Eriolepis section. The antiproliferative assay of the compounds isolated from their optimum sources, confirmed a dose-dependent inhibitory effect of the structures bearing the 4',7-epoxy moiety (balanophonin, picrasmalignan, desmethyl balanophonin, desmethyl picrasmalignan) against SW480 colon cancer cells, while those bearing the 4',7-dihydroxy motif (prebalanophonin, prepicrasmalignan) were inactive.

Comprehensive Drug Testing of Patient-derived Conditionally Reprogrammed Cells from Castration-resistant Prostate Cancer.

BACKGROUND: Technology development to enable the culture of human prostate cancer (PCa) progenitor cells is required for the identification of new, potentially curative therapies for PCa. OBJECTIVE: We established and characterized patient-derived conditionally reprogrammed cells (CRCs) to assess their biological properties and to apply these to test the efficacies of drugs. DESIGN, SETTING, AND PARTICIPANTS: CRCs were established from seven patient samples with disease ranging from primary PCa to advanced castration-resistant PCa (CRPC). The CRCs were characterized by genomic, transcriptomic, protein expression, and drug profiling. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: The phenotypic quantification of the CRCs was done based on immunostaining followed by image analysis with Advanced Cell Classifier using Random Forest supervised machine learning. Copy number aberrations (CNAs) were called from whole-exome sequencing and transcriptomics using in-house pipelines. Dose-response measurements were used to generate multiparameter drug sensitivity scores using R-statistical language. RESULTS AND LIMITATIONS: We generated six benign CRC cultures which all had an androgen receptor-negative, basal/transit-amplifying phenotype with few CNAs. In three-dimensional cell culture, these cells could re-express the androgen receptor. The CRCs from a CRPC patient (HUB.5) displayed multiple CNAs, many of which were shared with the parental tumor. We carried out high-throughput drug-response studies with 306 emerging and clinical cancer drugs. Using the benign CRCs as controls, we identified the Bcl-2 family inhibitor navitoclax as the most potent cancer-specific drug for the CRCs from a CRPC patient. Other drug efficacies included taxanes, mepacrine, and retinoids. CONCLUSIONS: Comprehensive cancer pharmacopeia-wide drug testing of CRCs from a CRPC patient highlighted both known and novel drug sensitivities in PCa, including navitoclax, which is currently being tested in clinical trials of CRPC. PATIENT SUMMARY: We describe an approach to generate patient-derived cancer cells from advanced prostate cancer and apply such cells to discover drugs that could be applied in clinical trials for castration-resistant prostate cancer.

Vitamin D supplementation affects the IGF system in men after acute exercise.

OBJECTIVE: Contradictory data between the Insulin-Like Growth Factor System (IGF) system and exercise may be due to alteration in IGF binding proteins. Vitamin D (D) deficiency has been related to muscle weakness and Insulin Like Growth Factor Binding Protein 3 (IGFBP3). A Vit. D and acute exercise merge is proposed to modify the IGF system. DESIGN: D insufficient and deficient men (39.0±8.6yo with serum D (25OH D) 20.0±7.7ng/mL) did 1h of stretching (ST), aerobic (AB), and resistance (RT) exercises, before and after 28d of 4000IU/d Vit. D3 (D, n=6) or Placebo (P, n=7). ST, a time/attention control visit, interchanged unreceptive movements. AB was moderate intensity treadmill walking. RT rotated moderate strength 50% 1-RM repetitions (15, 10) of squat, bench press, leg press, and lat pull down. Serum Total IGF1 (TIGF1), Insulin Like Growth Factor Binding Protein 1 (IGFBP1), and IGFBP3 were measured before (T1, fasting), immediately after (T2), and 2h post (T3) exercise. RESULTS: After ST, IGFBP3 was greater in the D group at T2 (2948, 2130ng/mL; p<0.03) and T3 (3087, 2212; p<0.02). During RT, TIGF1 decreased in the Placebo (P) group from T1 to T3 (151.4, 107.3ng/mL; p<0.05), while IGFBP1 increased in the D group from T1 to T3 (26.5, 96.2ng/mL; p<0.05). RT IGFBP3 was greater at T1, T2, and T3 in the D group (2932.5, 2110.7; p<0.03), (3163.9, 2392.5; p<0.04), and (3355.3, 2353.1; p<0.01). In AB, IGFBP3 was greater in the D group at T2 (3128.6, 2226.3.0; p<0.04) and T3 (2949.7, 2135.1; p<0.05). CONCLUSION: D supplementation amplified IGFBP3 after low or moderate activity which may increase the delivery of IGF1 to tissues. Resistance exercise with D not only increased IGFBP3 and IGFBP1 levels but also conserved TIGF1 levels, possibly shifting the IGF system for enriched muscle well-being.

Screening out irrelevant cell-based models of disease.

The common and persistent failures to translate promising preclinical drug candidates into clinical success highlight the limited effectiveness of disease models currently used in drug discovery. An apparent reluctance to explore and adopt alternative cell- and tissue-based model systems, coupled with a detachment from clinical practice during assay validation, contributes to ineffective translational research. To help address these issues and stimulate debate, here we propose a set of principles to facilitate the definition and development of disease-relevant assays, and we discuss new opportunities for exploiting the latest advances in cell-based assay technologies in drug discovery, including induced pluripotent stem cells, three-dimensional (3D) co-culture and organ-on-a-chip systems, complemented by advances in single-cell imaging and gene editing technologies. Funding to support precompetitive, multidisciplinary collaborations to develop novel preclinical models and cell-based screening technologies could have a key role in improving their clinical relevance, and ultimately increase clinical success rates.

Next-generation sequencing of common osteogenesis imperfecta-related genes in clinical practice.

Next generation sequencing (NGS) is a rapidly developing area in genetics. Utilizing this technology in the management of disorders with complex genetic background and not recurrent mutation hot spots can be extremely useful. In this study, we applied NGS, namely semiconductor sequencing to determine the most significant osteogenesis imperfecta-related genetic variants in the clinical practice. We selected genes coding collagen type I alpha-1 and-2 (COL1A1, COL1A2) which are responsible for more than 90% of all cases. CRTAP and LEPRE1/P3H1 genes involved in the background of the recessive forms with relatively high frequency (type VII and VIII) represent less than 10% of the disease. In our six patients (1-41 years), we identified 23 different variants. We found a total of 14 single nucleotide variants (SNV) in COL1A1 and COL1A2, 5 in CRTAP and 4 in LEPRE1. Two novel and two already well-established pathogenic SNVs have been identified. Among the newly recognized mutations, one results in an amino acid change and one of them is a stop codon. We have shown that a new full-scale cost-effective NGS method can be developed and utilized to supplement diagnostic process of osteogenesis imperfecta with molecular genetic data in clinical practice.

Comparison of the prevalence of malnutrition diagnosis in head and neck, gastrointestinal, and lung cancer patients by 3 classification methods.

BACKGROUND: Malnutrition is prevalent among patients within certain cancer types. There is lack of universal standard of care for nutrition screening and a lack of agreement on an operational definition and on validity of malnutrition indicators. OBJECTIVE: In a secondary data analysis, we investigated prevalence of malnutrition diagnosis with 3 classification methods using data from medical records of a National Cancer Institute-designated comprehensive cancer center. METHODS: Records of 227 patients hospitalized during 1998 with head and neck, gastrointestinal, or lung cancer were reviewed for malnutrition based on 3 methods: (1) physician-diagnosed malnutrition-related International Classification of Diseases, Ninth Revision codes; (2) in-hospital nutritional assessment summaries conducted by registered dietitians; and (3) body mass indexes (BMIs). For patients with multiple admissions, only data from the first hospitalization were included. RESULTS: Prevalence of malnutrition diagnosis ranged from 8.8% based on BMI to approximately 26% of all cases based on dietitian assessment. κ coefficients between any methods indicated a weak (κ = 0.23, BMI and dietitians; and κ = 0.28, dietitians and physicians)-to-fair strength of agreement (κ = 0.38, BMI and physicians). CONCLUSIONS: Available methods to identify patients with malnutrition in a National Cancer Institute-designated comprehensive cancer center resulted in varied prevalence of malnutrition diagnosis. A universal standard of care for nutrition screening that uses validated tools is needed. IMPLICATIONS FOR PRACTICE: The Joint Commission on the Accreditation of Healthcare Organizations requires nutritional screening of patients within 24 hours of admission. For this purpose, implementation of a validated tool that can be used by various healthcare practitioners, including nurses, needs to be considered.

Equol status modifies the association of soy intake and mammographic density in a sample of postmenopausal women.

Only 30% to 50% of people produce the daidzein-metabolite equol after eating soy. We conducted a cross-sectional study of the associations between equol status, intake of soy foods, and mammographic density in a sample of postmenopausal women recruited at a radiology clinic near Buffalo, New York. Participants were 48 to 82 years old, had no history of cancer or breast reduction/augmentation, and no recent use of antibiotics or hormones. Percent density was measured by computer-assisted analysis of digitized images of craniocaudal films. Equol status was assessed using a soy-challenge protocol and usual soy intake by questionnaire. General linear models were used to assess independent and joint effects of equol status and intake of soy on multivariate adjusted percent density (covariates included age, body mass index, parity, age at first birth, and ever use of combined hormone therapy). Of 325 enrolled, 232 (71%) participants completed study assessments and are included in the present analysis. Mean percent density was 34% (+/-18%). Seventy-five (30%) participants were producers of equol. Forty-three (19%) participants reported regularly eating >1 soy food or supplement/wk. There were no significant independent associations of equol status or soy intake with percent density, but the interaction between these factors was significant (P < 0.01). Among equol producers, those with weekly soy intake had lower percent density (30.7% in weekly consumers of soy versus 38.9% in others; P = 0.08); among nonproducers, weekly soy intake was associated with higher percent density (37.5% in weekly soy consumers versus 30.7% in others; P = 0.03). Results suggest that equol producers and nonproducers may experience different effects of dietary soy on breast tissue.

Predictors of change in calcium intake in postmenopausal women after osteoporosis screening.

Osteoporosis is a serious public health concern. Understanding the extent to which a bone density assessment affects change in dietary intake in postmenopausal women is needed. This study investigated whether results of bone density screening tests resulted in reported initiation or change in either dietary and/or supplemental calcium intake. Between 1997 and 2000, dual-energy X-ray absorptiometry (DXA) screening was conducted on 1468 postmenopausal women as part of an ancillary study of the Women's Health Initiative Observational Study in Buffalo, New York. One year after bone density testing, a questionnaire was sent to determine change in lifestyle behaviors and dietary intake. Participants included in this analysis were 923 Caucasian women who had not had a prior bone density screening test, reported no prior diagnosis of osteoporosis and were not taking medication (other than hormone therapy) for osteoporosis. Of these, according to WHO T-score criteria, 36% had osteoporosis, 48% had osteopenia, and 17% had normal bone density. Factors associated (P < 0.05) with increase in calcium intake in crude analyses included: BMI, follow-up consultation with a health care provider, and osteopenia or osteoporosis compared with normal T-score level. In multivariate adjusted analyses, both osteopenia [OR = 2.37, 95% CI (1.45-3.89); P = 0.001] and osteoporosis [OR = 3.86, 95% CI (2.30-6.46); P = <0.001] found on DXA were strong independent predictors of women's decision to start or increase calcium intake. This study provided evidence that the results of osteoporosis DXA screening influence postmenopausal women's decisions to increase calcium intake.