Therapeutical interference with the epigenetic landscape of germ cell tumors: a comparative drug study and new mechanistical insights

Background Type II germ cell tumors (GCT) are the most common solid cancers in males of age 15 to 35 years. Treatment of these tumors includes cisplatin-based therapy achieving high cure rates, but also leading to late toxicities. As mainly young men are suffering from GCTs, late toxicities play a major role regarding life expectancy, and the development of therapy resistance emphasizes the need for alternative therapeutic options. GCTs are highly susceptible to interference with the epigenetic landscape; therefore, this study focuses on screening of drugs against epigenetic factors as a treatment option for GCTs. Results We present seven different epigenetic inhibitors efficiently decreasing cell viability in GCT cell lines including cisplatin-resistant subclones at low concentrations by targeting epigenetic modifiers and interactors, like histone deacetylases (Quisinostat), histone demethylases (JIB-04), histone methyltransferases (Chaetocin), epigenetic readers (MZ-1, LP99) and polycomb-repressive complexes (PRT4165, GSK343). Mass spectrometry-based analyses of the histone modification landscape revealed effects beyond the expected mode-of-action of each drug, suggesting a wider spectrum of activity than initially assumed. Moreover, we characterized the effects of each drug on the transcriptome of GCT cells by RNA sequencing and found common deregulations in gene expression of ion transporters and DNA-binding factors. A kinase array revealed deregulations of signaling pathways, like cAMP, JAK-STAT and WNT. Conclusion Our study identified seven drugs against epigenetic modifiers to treat cisplatin-resistant GCTs. Further, we extensively analyzed off-target effects and modes-of-action, which are important for risk assessment of the individual drugs.

Are Drugs that Cause Dysbiosis Longitudinally Associated with Cognitive Scores, Cognitive Impairment, & Dementia?

Recent research has examined how the microbiome may influence cognitive outcomes; however, there is a paucity of research understanding how medication associated with dysbiosis may be associated with cognitive changes. This study used data from the Health and Retirement Study and the Prescription Drug Study subset for adults 51 and older (n=3,898). Continuous (0-27) and categorical (cognitively normal=12-27; cognitive impairment=7-11; and dementia=0-6) cognitive outcomes were used. Prescriptions utilized were proton pump inhibitors, antibiotics, selective serotonin reuptake inhibitors, tricyclic antidepressants, antipsychotics, antihistamines, and a summed dose-response measure. Linear mixed models (LMM) and generalized linear mixed models (GLMM) were used for continuous and binary outcomes. For the LMM model, the main effect for those taking one medication was insignificant; however, the interaction with time showed a significant decrease over time (β: -0.07; 95% confidence interval (CI): -0.14, 0.01). The mean cognitive score was lower for those taking two or more medications (β: -1.48; 95% CI: -2.70, -0.25), although the interaction with time was insignificant. GLMM results showed those taking two or medications had odds that are 612% larger (odds ratio (OR): 7.12; 95% CI: 3.03, 16.71) of going from cognitively healthy to dementia but the interaction with time showed decreased odds over time (OR: 0.92; 95% CI 0.86, 0.97). For cognitive impairment, those who took two or more medications had odds that were 45% larger (OR: 1.45; 95% CI: 1.05, 2.00) of going from cognitively healthy to cognitively impaired. This study indicated a dose-response aspect to taking medications on cognitive outcomes.

Challenges with Conducting an Investigational Drug Study in Older Adults in Nursing Homes During a Pandemic

In the early months of the pandemic, SARS-CoV-2 infected nursing home residents in explosive and deadly outbreaks. Nursing home residents disproportionately accounted for over 40% of COVID-19 mortality nationally. This national emergency drove scientific and public health experts to develop and implement administrative, clinical, and research programs to limit the pandemic’s impact, especially for high-risk individuals, such as those hospitalized or living in nursing homes. Nursing home policies, prompted by the Centers for Medicare & Medicaid Services (CMS) and the Centers for Disease Control and Prevention (CDC) guidance, severely restricted access beginning in March 2020 in an effort to limit disease exposure. In July 2020 we began the process to conduct an investigational SARS-CoV-2 post exposure prophylaxis study of nursing home residents, incorporating FDA guidance developed for conducting investigational drug trials in the context of COVID-19. Our research teams adapted our nursing home engagement, resident consenting and research data collection strategies accordingly. We remotely screened residents living in any of 28 nursing homes for eligibility to participate, ultimately consenting and randomizing individuals in 11 facilities. Of the 2,683 nursing home residents 65 years or older we screened, 48 (1.8%) agreed to consent individually or through proxy, most often a legally authorized representative. We will describe our research methods, with emphasis on how we addressed challenges presented due to performing all research tasks remotely and identify strategies that can qualitatively improve the remote nursing home research experience.

Detection of biofilm formation, extended spectrum beta lactamase production and their correlation with antibiotic resistance among uropathogenic Escherichia coli

Urinary tract infections (UTI) are most common bacterial infections encountered in clinical practice and is predominant organism causing UTI.: 1.To study antibiotic resistance pattern among uropathogenic (UPEC). 2.To study extended spectrum beta lactamase (ESBL) production and their correlation with antibiotic resistance among uropathogenic (UPEC). 3.To study biofilm formation and its correlation with antibiotic resistance among uropathogenic (UPEC)A prospective study was conducted on first 100 isolated from urine specimens of patients suspected to be having urinary tract infection between January 2016 and December 2016 received at Department of Microbiology, SIMS Shimoga. Fresh midstream urine samples were aseptically collected in sterile containers and plated on Blood agar & MacConkey agar plates using a standard loop technique & the growth was processed by standard bacteriological technique. Biofilm detection was done by tube and microtitre plate method. ESBL detection was done according to CLSI criteria. Antimicrobial sensitivity testing was done using Kirby-Bauer methods on Mueller-Hinton agar. Results were interpreted as per the CLSI guidelines.Antibiotic sensitivity of was nitrofurantoin (100%) fosfomycin (100%) imipenem (77%) cotrimoxazole (61%) amikacin (47%) aztreonam (53%) piperacillintazobactam (41%). cefotaxim (25%) ceftazidime (32%) norfloxacin (20%) ampicillin-sulbactam (12%) ciprofloxacin (15%), levofloxacin (12%), amoxiclavulanicacid (15%). ESBL positive were 83% (screening), 80% (confirmatory). Biofilm positive were 63% (tube method) 79% (Microtiterplate method).This study showed Fosfomycin and Nitrofurantoin were the most sensitive drug. Study shows antibiotic resistance was seen more in ESBL and Biofilm producers compared to non ESBL and non biofilm producers.

Sales of antibiotics and hydroxychloroquine in India during the COVID-19 epidemic: An interrupted time series analysis

Background We assessed the impact of the coronavirus disease 2019 (COVID-19) epidemic in India on the consumption of antibiotics and hydroxychloroquine (HCQ) in the private sector in 2020 compared to the expected level of use had the epidemic not occurred. Methods and findings We performed interrupted time series (ITS) analyses of sales volumes reported in standard units (i.e., doses), collected at regular monthly intervals from January 2018 to December 2020 and obtained from IQVIA, India. As children are less prone to develop symptomatic severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, we hypothesized a predominant increase in non-child-appropriate formulation (non-CAF) sales. COVID-19-attributable changes in the level and trend of monthly sales of total antibiotics, azithromycin, and HCQ were estimated, accounting for seasonality and lockdown period where appropriate. A total of 16,290 million doses of antibiotics were sold in India in 2020, which is slightly less than the amount in 2018 and 2019. However, the proportion of non-CAF antibiotics increased from 72.5% (95% CI: 71.8% to 73.1%) in 2019 to 76.8% (95% CI: 76.2% to 77.5%) in 2020. Our ITS analyses estimated that COVID-19 likely contributed to 216.4 million (95% CI: 68.0 to 364.8 million; P = 0.008) excess doses of non-CAF antibiotics and 38.0 million (95% CI: 26.4 to 49.2 million; P < 0.001) excess doses of non-CAF azithromycin (equivalent to a minimum of 6.2 million azithromycin treatment courses) between June and September 2020, i.e., until the peak of the first epidemic wave, after which a negative change in trend was identified. In March 2020, we estimated a COVID-19-attributable change in level of +11.1 million doses (95% CI: 9.2 to 13.0 million; P < 0.001) for HCQ sales, whereas a weak negative change in monthly trend was found for this drug. Study limitations include the lack of coverage of the public healthcare sector, the inability to distinguish antibiotic and HCQ sales in inpatient versus outpatient care, and the suboptimal number of pre- and post-epidemic data points, which could have prevented an accurate adjustment for seasonal trends despite the robustness of our statistical approaches. Conclusions A significant increase in non-CAF antibiotic sales, and particularly azithromycin, occurred during the peak phase of the first COVID-19 epidemic wave in India, indicating the need for urgent antibiotic stewardship measures.

Evaluation of the Adding Paracetamol to Dexmedetomidine in Pain Management After Adult Cardiac Surgery

Background: Postoperative pain control after cardiac surgery is usually based on Opioids. These drugs are associated with side effects, and the use of drugs with fewer side effects is important for analgesia. Dexmedetomidine and paracetamol have fewer side effects than opioids. Objectives: The aim of the study was to evaluate the adding paracetamol to dexmedetomidine continuous infusion pump for pain management after adult cardiac surgery. Methods: In this study, 100 patients were divided into two groups. One group received a continuous infusion of dexmedetomidine and paracetamol (DP), and the other received dexmedetomidine (D). These two groups were evaluated for MAP, HR, and the need for prescribing opioids before and after extubation. Also, duration of intubation and pain before extubation and 36 hours after every 4 hours. Results: Patients in the DP group had lower mean MAP and HR during intubation period than the D group and needed fewer opioids and doses of opioids in addition to drug study infusion pre- (P = 0.001) and post-extubation (P = 0.001 and P = 0.022, respectively). The DP group patients were extubated earlier (P = 0.001). After extubation, the DP group had less pain than the D group. Conclusions: This study showed that adding paracetamol to the dexmedetomidine infusion pump can provide better analgesia.

A Dual Target-Directed Single Domain-Based Fusion Protein Against Interleukin-6 Receptor Decelerate Experimental Arthritis Progression Via Modulating JNK Expression

The currently used anti-cytokine therapeutic antibodies cannot selectively neutralize pathogenic cytokine signalling that cause collateral damage to protective signalling cascades. The single domain chain firstly discovered in Camelidae displays fully functional ability in antigen-binding against variable targets, which has been seemed as attractive candidates for the next-generation biologic drug study. In this study, we established a simple prokaryotic expression system for a dual target-directed single domain-based Fusion Protein against the interleukin-6 receptor and human serum, albumin, the recombinant anti-IL-6R fusion protein (VHH-0031). VHH-0031 exhibited potent anti-inflammatory effects produced by LPS on cell RAW264.7, where the major cytokines and NO production were down-regulated after 24 hr incubation with VHH-0031 in a dose-dependent manner. In vivo, VHH-0031 presented significant effects on the degree reduction of joint swelling in the adjuvant-induced arthritis (AIA) rat, having a healthier appearance compared to the dexamethasone. The expression level of JNK protein in the VHH-0031 group was significantly decreased, demonstrating that VHH-0031 provides a low-cost and desirable effect in the treatment of more widely patients.

Molecular Analysis of IL-5 Receptor Subunit Alpha as a Possible Pharmacogenetic Biomarker in Asthma

Background: Asthma is a heterogeneous syndrome with a broad clinical spectrum and high drug response variability. The inflammatory response in asthma involves multiple effector cells and mediator molecules. Based on asthma immunopathogenesis, precision medicine can be a promising strategy for identifying biomarkers. Biologic therapies acting on the IL-5/IL-5 receptor axis have been developed. IL-5 promotes proliferation, differentiation and activation of eosinophils by binding to the IL-5 receptor, located on the surface of eosinophils and basophils. This study aimed to investigate the expression of IL5RA in patients with several types of asthma and its expression after treatment with benralizumab, a biologic directed against IL-5 receptor subunit alpha.Methods: Sixty peripheral blood samples, 30 from healthy controls and 30 from asthmatic patients, were selected for a transcriptomic RNAseq study. Differential expression analysis was performed by statistical assessment of fold changes and P-values. A validation study of IL5RA expression was developed using qPCR in 100 controls and 187 asthmatic patients. The effect of benralizumab on IL5RA expression was evaluated in five patients by comparing expression levels between pretreatment and after 3 months of treatment. The IL5RA mRNA levels were normalized to GAPDH and TBP expression values for each sample. Calculations were made by the comparative ΔΔCt method. All procedures followed the MIQE guidelines.Results:IL5RA was one of the most differentially overexpressed coding transcripts in the peripheral blood of asthmatic patients (P = 8.63E-08 and fold change of 2.22). In the qPCR validation study, IL5RA expression levels were significantly higher in asthmatic patients than in controls (P &lt; 0.001). Significant expression differences were present in different asthmatic types. In the biological drug study, patients treated with benralizumab showed a significant decrease in IL5RA expression and blood eosinophil counts. A notable improvement in ACT and lung function was also observed in these patients.Conclusions: These results indicate that IL5RA is overexpressed in patients with different types of asthma. It could help identify which asthmatic patients will respond more efficiently to benralizumab, moving toward a more personalized asthma management. Although further studies are required, IL5RA could play a role as a biomarker and pharmacogenetic factor in asthma.

DrugWAS: Leveraging drug-wide association studies to facilitate drug repurposing for COVID-19

ImportanceThere is an unprecedented need to rapidly identify safe and effective treatments for the novel coronavirus disease 2019 (COVID-19).ObjectiveTo systematically investigate if any of the available drugs in Electronic Health Record (EHR), including prescription drugs and dietary supplements, can be repurposed as potential treatment for COVID-19.Design, Setting, and ParticipantsBased on a retrospective cohort analysis of EHR data, drug-wide association studies (DrugWAS) were performed on COVID-19 patients at Vanderbilt University Medical Center (VUMC). For each drug study, multivariable logistic regression with overlap weighting using propensity score was applied to estimate the effect of drug exposure on COVID-19 disease outcomes.ExposuresPatient exposure to a drug during 1-year prior to the pandemic and COVID-19 diagnosis was chosen as exposure of interest. Natural language processing was employed to extract drug information from clinical notes, in addition to the prescription drug data available in structured format.Main Outcomes and MeasuresAll-cause of death was selected as primary outcome. Hospitalization, admission to the intensive care unit (ICU), and need for mechanical ventilation were identified as secondary outcomes.ResultsThe study included 7,768 COVID-19 patients, of which 509 (6.55%) were hospitalized, 82 (1.06%) were admitted to ICU, 64 (0.82%) received mechanical ventilation, and 90 (1.16%) died. Overall, 15 drugs were significantly associated with decreased COVID-19 severity. Previous exposure to either Streptococcus pneumoniae vaccines (adjusted odds ratio [OR], 0.38; 95% CI, 0.14-0.98), diphtheria toxoid vaccine (OR, 0.39; 95% CI, 0.15-0.98), and tetanus toxoid vaccine (OR, 0.39; 95% CI, 0.15-0.98) were significantly associated with a decreased risk of death (primary outcome). Secondary analyses identified several other significant associations showing lower risk for COVID-19 outcomes: 2 vaccines (acellular pertussis, Streptococcus pneumoniae), 3 dietary supplements (turmeric extract, flaxseed extract, omega-3 fatty acids), methylprednisolone acetate, pseudoephedrine, ethinyl estradiol, estradiol, ibuprofen, and fluticasone.Conclusions and RelevanceThis cohort study leveraged EHR data to identify a list of drugs that could be repurposed to improve COVID-19 outcomes. Further randomized clinical trials are needed to investigate the efficacy of the proposed drugs.Key PointsQuestionCan Electronic Health Records (EHRs) be used to search for drug candidates that could be repurposed to treat the coronavirus disease 2019 (COVID-19)?FindingsDrug-wide association studies (DrugWAS) of COVID-19 severity outcomes were conducted on a cohort of 7,768 COVID-19 patients. The study found 15 drug ingredients that are significantly associated with a decreased risk of death and other severe COVID-19 outcomes.MeaningThe list of drugs proposed by this study could provide additional insights into developing new candidates for COVID-19 treatment.