The Efficacy of Nitric Oxide-Generating Lozenges on Outcome in Newly Diagnosed COVID-19 Patients of African American and Hispanic Origin.

BACKGROUND: The study was initiated in 2020 to test the efficacy of a nitric oxide-generating lozenge (NOL) in outpatients with newly diagnosed COVID-19 to mitigate disease severity. The study enrolled high-risk patients, African American and Latino. METHODS: This was a randomized, double-blinded, prospective, placebo-controlled trial. The primary endpoint was hospitalization, intensive care unit admission, intubation, dialysis, and death. The secondary endpoints were time to symptom resolution and the effect on oxygen saturation. Patients ages 50-85 years with recent COVID-19 diagnosis with at least one risk factor were recruited. Patients were randomized to either active treatment or placebo using block randomization. Blood pressure and oxygen saturation (SpO2) was measured prior to and after the first dose and each morning thereafter. RESULTS: A total of 840 patients was planned, half in each of the lozenge and placebo groups. An interim review of data was prespecified. Of 524 patients, the composite endpoint occurred in 6 patients, 3 (1.1%) in each group. The time to symptom resolution was 1 day shorter on active treatment (8.7 ± 6.6 vs 9.8 ± 6.8 days) (P = .3). There was no change in SpO2 on placebo (0.0 ± 2.0%) and no significant change on treatment (0.14 ± 0.9%), P = .3. All events occurred in the first year (2020). CONCLUSIONS: This study did not find a benefit of NOL therapy in COVID-19 patients and was terminated for futility. NOL treatment did not reduce mortality, hospitalization, intubation, or a reduction in symptoms duration. The study did find the NO lozenges were well tolerated in high-risk patients, without reported side effects.

Ancestral Stem Cell Reprogramming Genes Active in Hemichordate Regeneration.

Hemichordate enteropneust worms regenerate extensively in a manner that resembles the regeneration for which planaria and hydra are well known. Although hemichordates are often classified as an extant phylogenetic group that may hold ancestral deuterostome body plans at the base of the deuterostome evolutionary line leading to chordates, mammals, and humans, extensive regeneration is not known in any of these more advanced groups. Here we investigated whether hemichordates deploy functional homologs of canonical Yamanaka stem cell reprogramming factors, Oct4, Sox2, Nanog, and Klf4, as they regenerate. These reprogramming factors are not expressed during regeneration of limbs, fins, eyes or other structures that represent the best examples of regeneration in chordates. We first examined Ptychodera flava EST libraries and identified Pf-Pou3, Pf-SoxB1, Pf-Msxlx, and Pf-Klf1/2/4 as most closely related to the Yamanaka factors, respectively. In situ hybridization analyses revealed that all these homologs are expressed in a distinct manner during head regeneration. Furthermore, Pf-Pou3 partially rescued the loss of endogenous Oct4 in mouse embryonic stem cells in maintaining the pluripotency gene expression program. Based on these results, we propose that hemichordates may have co-opted these reprogramming factors for their extensive regeneration or that chordates may have lost the ability to mobilize these factors in response to damage. The robustness of these pluripotency gene circuits in the inner cell mass and in formation of induced pluripotent stem cells from mammalian somatic cells shows that these programs are intact in humans and other mammals and that these circuits may respond to as yet unknown gene regulatory signals, mobilizing full regeneration in hemichordates.

Infectious Bronchitis Hatchery Vaccination: Comparison between Traditional Spray Administration and a Newly Developed Gel Delivery System in Field Conditions.

The control of infectious bronchitis (IB) is essential in intensive broiler production and is pursued through strict biosecurity and mass vaccination. Despite effective and routinely adopted, hatchery spray vaccination has been hypothesized to affect chicks' body temperature and wellbeing. Recently, gel administration has been proposed as an alternative and proved feasible in experimental settings. In this study, IBV spray and gel vaccination methods were compared in field conditions. One hundred birds from the same hatch were enrolled in the study and vaccinated, half by spray and half by gel, with 793B and Mass vaccines. After vaccination, rectal temperature was measured and vaccine intake assessed. The two groups were housed for 35 days in separate pens and swabs and blood samples were collected at multiple time points for genotype-specific molecular analyses and serology, respectively. The temperature was significantly lower in spray-vaccinated chicks 10 min and an hour after administration. A similar trend in 793B titres was observed in both groups, while the Mass vaccine was detected later but persisted longer in gel-vaccinated chicks. No differences were observed in mean antibody titres. Compared to spray, gel administration appears equally effective and less impactful on body temperature, thus supporting its application for IBV vaccination.

A Study of Proton Pump Inhibitors and Other Risk Factors in Warfarin-Associated Gastrointestinal Bleeding.

Background Warfarin users are at increased risk for gastrointestinal bleeding (GIB). History of GIB, stroke, cardiovascular or chronic kidney disease, age greater than 65 years, and drug interaction with proton pump inhibitors (PPI) have previously been identified as risk factors for GIB in warfarin users. We hypothesized that concomitant use of warfarin and PPI would increase the incidence of GIB relative to warfarin use alone. Methods We did a retrospective review of medical records of 626 patients taking warfarin for at least two weeks. Parameters including age, concomitant medication use (non-steroidal anti-inflammatory drugs (NSAID), aspirin, selective serotonin reuptake inhibitors (SSRIs), PPI, and anti-platelet drug), history of GIB, chronic renal failure (CRF), and peptic ulcer disease (PUD) prior to warfarin use were analyzed. Results Variables that increase the likelihood of bleeding in warfarin users included aspirin, PPI, history of PUD, history of previous GIB, CRF, and elevated prothrombin time (PT)/international normalized ratio (INR) values. Concomitant antiplatelet use showed a slight increase in GIB but this was not statistically significant (p=0.082). NSAID use and SSRI use were not associated with a higher risk of GIB among warfarin users. Patients who are on PPI and warfarin simultaneously are more likely to be on acetylsalicylic acid (ASA) or have a history of PUD, GIB, or CRF, all of which are associated with increased incidences of GIB. Conclusions Although concomitant use of warfarin and PPI appears to be associated with an increased incidence of GIB, these patients are more likely to have other risk factors that also increase the risk of a GIB outcome. Therefore, the interaction between PPI and warfarin is clinically insignificant.

Modular Reorganization of Signaling Networks during the Development of Colon Adenoma and Carcinoma.

Network science is an emerging tool in systems biology and oncology, providing novel, system-level insight into the development of cancer. The aim of this project was to study the signaling networks in the process of oncogenesis to explore the adaptive mechanisms taking part in the cancerous transformation of healthy cells. For this purpose, colon cancer proved to be an excellent candidate as the preliminary phase, and adenoma has a long evolution time. In our work, transcriptomic data have been collected from normal colon, colon adenoma, and colon cancer samples to calculating link (i.e., network edge) weights as approximative proxies for protein abundances, and link weights were included in the Human Cancer Signaling Network. Here we show that the adenoma phase clearly differs from the normal and cancer states in terms of a more scattered link weight distribution and enlarged network diameter. Modular analysis shows the rearrangement of the apoptosis- and the cell-cycle-related modules, whose pathway enrichment analysis supports the relevance of targeted therapy. Our work enriches the system-wide assessment of cancer development, showing specific changes for the adenoma state.

A Novel Experimental Study on the Rheological Properties and Thermal Conductivity of Halloysite Nanofluids.

Nanofluids obtained from halloysite and de-ionized water (DI) were prepared by using surfactants and changing pH for heat-transfer applications. The halloysite nanotubes (HNTs) nanofluids were studied for several volume fractions (0.5, 1.0, and 1.5 vol%) and temperatures (20, 30, 40, 50, and 60 °C). The properties of HNTs were studied with a scanning electron microscope (SEM), energy-dispersive X-ray analysis (EDX), Fourier-transform infrared (FT-IR) spectroscopy, X-ray powder diffraction (XRD), Raman spectroscopy and thermogravimetry/differential thermal analysis (TG/DTA). The stability of the nanofluids was proven by zeta potentials measurements and visual observation. With surfactants, the HNT nanofluids had the highest thermal conductivity increment of 18.30% for 1.5 vol% concentration in comparison with the base fluid. The thermal conductivity enhancement of nanofluids containing surfactant was slightly higher than nanofluids with pH = 12. The prepared nanofluids were Newtonian. The viscosity enhancements of the nanofluid were 11% and 12.8% at 30 °C for 0.5% volume concentration with surfactants and at pH = 12, respectively. Empirical correlations of viscosity and thermal conductivity for these nanofluids were proposed for practical applications.

Model-Informed Development of Sotalol Loading and Dose Escalation Employing an Intravenous Infusion.

BACKGROUND: Sotalol is often employed to prevent recurrence of symptomatic atrial flutter/atrial fibrillation. Because sotalol can prolong the QT interval excessively causing ventricular arrhythmias, a 3-day in-hospital loading or dose escalation period is mandated with oral administration in the product label for patient safety. In patients with normal renal function, 3 days (five oral doses) are required to obtain steady state maximum sotalol concentration, which results in maximum QT prolongation. The aim of this study is to develop an intravenous to oral loading regime for sotalol therapy that reduces the 3-day in-hospital initiation or dose escalation with oral administration to 1 day without compromising patient safety. METHODS: Using model-informed drug development techniques, simulations were developed for initiation and dose escalation of sotalol therapy by employing an intravenous loading dose followed by oral sotalol administrations. RESULTS: In patients with normal renal function, an initial 1-h loading dose of intravenous sotalol followed by two oral doses in 24 h has been developed permitting attainment of three maximum serum concentrations reflecting maximum QT prolongation in a 1-day observation period. Dosing regimens for patients with impaired renal function are also developed. CONCLUSIONS: In patients with normal renal function, using an intravenous loading dose followed by oral administrations permits safe initiation or dose escalation of sotalol in 1 day instead of the 3-day dosing regimen with oral administration.

Inhibition of Human Ether-A-Go-Go-Related Gene (hERG) Potassium Current by the Novel Sotalol Analogue, Soestalol.

The clinical utility of intravenous sotalol is limited due to an extended half-life combined with the potential to generate life-threatening arrhythmias. The authors developed a novel sotalol analogue, soestalol, with an ester linkage introduced to the molecule to shorten half-life. Their hypothesis was that soestalol, but not the acid metabolite, would inhibit the hERG potassium current. Whole-cell, voltage-clamp experiments were performed on cells expressing hERG. Soestalol inhibited outward IhERG tail current density in a manner similar to conventional sotalol. Additionally, soestalol right shifted the voltage dependence of activation. These results warrant further assessment of soestalol as a short-acting, Class III antiarrhythmic drug.

What is New in Pharmacologic Therapy for Cardiac Resuscitation?

Antiarrhythmic therapy can be a critical component of cardiac resuscitation. Therapies in this area have seen little advance in the last decade. Bretylium, a very old drug, has been reintroduced for ventricular tachycardia/ventricular fibrillation (VT/VF) therapy. There are still important questions to be addressed with bretylium: when to administer (first- or second-line) and at which dose. These questions and the development of newer agents will be areas of future research.

Identification of a novel archaea virus, detected in hydrocarbon polluted Hungarian and Canadian samples.

Metagenomics is a helpful tool for the analysis of unculturable organisms and viruses. Viruses that target bacteria and archaea play important roles in the microbial diversity of various ecosystems. Here we show that Methanosarcina virus MV (MetMV), the second Methanosarcina sp. virus with a completely determined genome, is characteristic of hydrocarbon pollution in environmental (soil and water) samples. It was highly abundant in Hungarian hydrocarbon polluted samples and its genome was also present in the NCBI SRA database containing reads from hydrocarbon polluted samples collected in Canada, indicating the stability of its niche and the marker feature of this virus. MetMV, as the only currently identified marker virus for pollution in environmental samples, could contribute to the understanding of the complicated network of prokaryotes and their viruses driving the decomposition of environmental pollutants.

Complete Genome Sequences of 10 Xanthomonas oryzae pv. oryzae Bacteriophages.

Xanthomonas oryzae pv. oryzae is the causative agent of bacterial leaf blight of rice. The application of bacteriophages may provide an effective tool against this bacterium. Here, we report the complete genome sequences of 10 newly isolated OP2-like X. oryzae pv. oryzae bacteriophages.

Small RNA NGS Revealed the Presence of Cherry Virus A and Little Cherry Virus 1 on Apricots in Hungary.

Fruit trees, such as apricot trees, are constantly exposed to the attack of viruses. As they are propagated in a vegetative way, this risk is present not only in the field, where they remain for decades, but also during their propagation. Metagenomic diagnostic methods, based on next generation sequencing (NGS), offer unique possibilities to reveal all the present pathogens in the investigated sample. Using NGS of small RNAs, a special field of these techniques, we tested leaf samples of different varieties of apricot originating from an isolator house or open field stock nursery. As a result, we identified Cherry virus A (CVA) and little cherry virus 1 (LChV-1) for the first time in Hungary. The NGS results were validated by RT-PCR and also by Northern blot in the case of CVA. Cloned and Sanger sequenced viral-specific PCR products enabled us to investigate their phylogenetic relationships. However, since these pathogens have not been described in our country before, their role in symptom development and modification during co-infection with other viruses requires further investigation.

Use of siRNAs for Diagnosis of Viruses Associated to Woody Plants in Nurseries and Stock Collections.

Woody perennial plants like grapevine and fruit trees can be infected by several viruses even as multiple infections. Since they are propagated vegetatively, the phytosanitary status of the propagation material (both the rootstock and the variety) can have a profound effect on the lifetime and health of the new plantations. The fast evolution of sequencing techniques provides a new opportunity for metagenomics-based viral diagnostics. Viral derived small RNAs produced by the host immune system during viral infection can be sequenced by next-generation techniques and analyzed for the presence of viruses, revealing the presence of all known viral pathogens in the sample. This method is based on Illumina sequencing of short RNAs and bioinformatics analysis of virus-derived small RNAs in the host. Here we describe a protocol for this challenging technique step by step with notes, in order to ensure success for every user.

Application of Targeted Next-generation Sequencing, TruSeq Custom Amplicon Assay for Molecular Pathology Diagnostics on Formalin-fixed and Paraffin-embedded Samples.

The implementation of targeted therapies revolutionized oncology. As the number of new oncogenic driver mutations, which provide molecular targets for prediction of effective and selective therapies, is increasing, the implementation of fast and reliable methods by molecular pathology labs is very important. Here we report our results with TruSeq Custom Amplicon assay performed on formalin-fixed and paraffin-embedded material. The oligo capture probes targeted the hotspot regions of 10 well-known oncogenes linked to clinical diagnosis and treatment of lung and colorectal adenocarcinomas, melanomas, and gastrointestinal stromal tumors. Fifteen previously genotyped formalin-fixed and paraffin-embedded DNA samples from different tumor types were selected for massively parallel sequencing. A bioinformatics pipeline was developed to identify high-quality variants and remove sequence artifacts. With the exception of 1 sample, which was of lower quality than the others, relevant mutations corresponding to tumor types could be reliable detected by the developed bioinformatical pipeline. This study indicates that the application of TruSeq Custom Amplicon assay is a promising tool in molecular pathology diagnostics, but it is important to standardize sample processing (including fixation, isolation procedure, sample selection based on quality assessment, and rigorous variant calling) to achieve the highest success rate and avoid false results.

Adherence to Evidence-Base Endotracheal Intubation Practice Patterns by Intensivists and Emergency Department Physicians.

BACKGROUND: Endotracheal intubation outside the operating room (OR) is mainly performed by intensive care (IC) physicians and emergency department (ED) physicians. We hypothesized that difference in practice patterns exists between these two groups of physicians. METHODS: A retrospective chart review was performed on all endotracheal intubations that were performed out of OR over a fi ve year period at our health care facility. Practice patterns of IC and ED physicians were compared regarding use of (a) video laryngoscopy, (b) paralytic agents, (c) waveform capnography, and (d) use of larger size of endotracheal tube (internal diameter ≥ 8 mm). RESULTS: A total of 201 patients underwent out of OR intubations over a 5 year period. IC physicians used more often than ED physicians video laryngoscopy (67% vs. 49%; p = 0.008), waveform capnography (99% vs. 86%; p = 0.001) and larger size endotracheal tubes (95% vs. 60%; p < 0.001). Conversely, paralytic agents were used less frequently by IC than ED physicians (12% vs. 51%; p < 0.001). The success of fi rst intubation attempt was higher by IC than ED physicians (82% vs. 67%; p = 0.018). CONCLUSIONS: IC physicians more often adhered to currently considered preferable practices for endotracheal intubation than ED physicians in this single center retrospective study. Although larger scale studies are needed to unveil the effects of different practice patterns on short and long term outcomes, the present study identifi es opportunity to bridge practice gaps that could lead to improved outcomes.

The full mitochondrial genomes of Mangalica pig breeds and their possible origin.

The mitogenomes of one animal of each of the three Mangalica breeds, Blonde, Red, and Swallow-belly were assembled from reads obtained by Next Generation Sequencing of the three genomes. Features of the mitogenomes were identical in the three breeds, apart from a second tRNA-Val gene on the L strand in Swallow-belly. Phylogenetic comparison of the three mitogenomes with 112 full mtDNA sequences clearly put Mangalicas into the European clade. Comparing the mitogenome of eight Mangalica animals revealed particular differences between them. The mitogenome of some Mangalicas was closely related to the Croatian Turopolje breed and this indicates either the common origin of their maternal lineages or admixture of some populations of the breeds. However, the origin of the mitogenome of certain purebred Mangalicas kept in the Hungarian Mangalica Gene Reserve still remains unknown.

Profile of Acute Asthma Exacerbation in Drug Users.

The characteristics of patients who use heroin, cocaine, or both and present with acute asthma exacerbations have not been well studied. In this retrospective study, we aimed to study the demographic characteristics of this patient population, the characteristics of their asthma attack, and the risk factors for the need for invasive mechanical ventilation in this patient population. We reviewed the charts of patients discharged from an inner-city hospital with a diagnosis of acute asthma exacerbation. Individuals who used either heroin or cocaine or both within 24 hours of presenting to the emergency department were identified as a cohort of drug users. The rest were classified as non-drug users. Both groups were compared, and a univariate analysis was performed. To assess the predictive value of drug use for the need for intubation in the presence of confounding factors, logistic regression analysis was performed to identify whether using cocaine or heroin or both was an individual predictor for the need for invasive ventilation. Data from 218 patients were analyzed. Drug users (n = 85) were younger (mean age in years 43.9 vs. 50.5, P < 0.01), predominantly male (63.5% vs. 33.8%, P < 0.01), and more likely to be cigarette smokers (90.6% vs. 57.6%, P < 0.01). A medical history of intubation and admissions to the intensive care unit (ICU) was more common among drug users (56.5% vs. 29.3%, P < 0.01 and 54.1% vs. 38.3%, P < 0.03, respectively). Drug use was associated with increased need for invasive mechanical ventilation (35% vs. 23.3%, P = 0.05). Non-drug users were more likely to be using inhaled corticosteroids (48.9% vs. 32.9%, P = 0.03) and had longitudinal care established with a primary care provider (50.6% vs. 68.9%, P < 0.01). After adjusting for a history of mechanical ventilation, history of ICU admission, use of systemic corticosteroids, smoking, and acute physiological assessment and chronic health evaluation 2 score, drug use remained predictive for the need for mechanical ventilation (P = 0.026). Acute asthma exacerbations triggered by cocaine and heroin should be treated aggressively because they represent a cohort with poor follow-up and undertreated asthma as outpatients and are associated with increased need for invasive mechanical ventilation and ICU admission during acute exacerbation.

Efficacy of Intravenous and Oral Sotalol in Pharmacologic Conversion of Atrial Fibrillation: A Systematic Review and Meta-Analysis.

OBJECTIVES: The role of sotalol is well established for the maintenance of sinus rhythm after successful conversion of atrial fibrillation (AF). However, its role in pharmacologic conversion of AF is poorly defined. The purpose of this study is to compare the efficacy of sotalol to that of other antiarrhythmic agents for AF conversion. METHODS: Standard methods of meta-analysis were employed. Full-text publications of clinical trials in English that compared the efficacy of sotalol to that of other antiarrhythmics or placebo/no treatment were eligible for inclusion. RESULTS: A systematic review revealed 10 eligible publications. Sotalol was superior to placebo and/or no antiarrhythmic therapy in AF conversion, with a relative success of 24 (95% CI 4.7-119, p < 0.001). Sotalol was not significantly different from class IA antiarrhythmic drugs. Similarly, sotalol was not different from class IC antiarrhythmic drugs or amiodarone in terms of conversion efficacy. In one study, sotalol was less effective than high-dose ibutilide (2 mg), with a relative success of 0.248 (95% CI 0.128-0.481, p < 0.001). Ibutilide caused more proarrhythmia. CONCLUSIONS: Sotalol is as effective as class IA and class IC antiarrhythmic agents, and it is also as effective as amiodarone for pharmacologic conversion of AF. Only ibutilide at a high dose showed a greater conversion rate of AF.

Sotalol versus Amiodarone in Treatment of Atrial Fibrillation.

The availability of intravenous (IV) Sotalol has equalized the treatment options since both amiodarone and sotalol are available in both IV and oral formulations. A review of the efficacy of sotalol as compared to amiodarone both for conversion of atrial fibrillation (AF) and maintenance of normal sinus rhythm (NSR) following cardiac surgery was undertaken. Standard methods of meta-analysis were employed. Full text publications of clinical trials written in English that compared the efficacy of sotalol to amiodarone were included in the analysis. For the conversion of AF to NSR, five studies were found eligible for the analysis. Two studies clinically compared sotalol to amiodarone for the maintenance of NSR after cardiac surgery. The common relative success of sotalol was 0.947 (95Cl: 0.837 to 1.071, P = 0.385), revealing essentially no differences in efficacy for conversion between amiodarone and sotalol. The average conversion rate was 47% with sotalol and 52% with amiodarone. The conversion rates were lower for persistent AF (sotalol 22% and amiodarone 27%), while greatest for recent onset AF (88% sotalol and 77% for amiodarone). The risk of developing post-operative atrial fibrillation was practically the same in both regimes, relative risk = 1.214 (95% CI: 0.815-1.808, p=0.339). In summary, sotalol and amiodarone are equally effective in AF conversion and maintenance of NSR post-cardiac surgery.