Bepridil is potent against SARS-CoV-2 in vitro.

Guided by a computational docking analysis, about 30 Food and Drug Administration/European Medicines Agency (FDA/EMA)-approved small-molecule medicines were characterized on their inhibition of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) main protease (Mpro). Of these small molecules tested, six displayed a concentration that inhibits response by 50% (IC50) value below 100 µM in inhibiting Mpro, and, importantly, three, that is, pimozide, ebastine, and bepridil, are basic molecules that potentiate dual functions by both raising endosomal pH to interfere with SARS-CoV-2 entry into the human cell host and inhibiting Mpro in infected cells. A live virus-based modified microneutralization assay revealed that bepridil possesses significant anti-SARS-CoV-2 activity in both Vero E6 and A459/ACE2 cells in a dose-dependent manner with low micromolar effective concentration, 50% (EC50) values. Therefore, the current study urges serious considerations of using bepridil in COVID-19 clinical tests.

The influence of labor epidural analgesia on maternal, uteroplacental and fetoplacental hemodynamics in normotensive parturients: a prospective observational study.

BACKGROUND: Epidural analgesia provides sufficient analgesia during labor but can cause hypotension despite various prophylactic measures. We studied its effects on pre-placental, fetoplacental, and fetal hemodynamics using Doppler ultrasound. The primary endpoint was the pulsatility index of the umbilical artery at 30 min after establishing epidural analgesia. Secondary endpoints included maternal blood pressures and neonatal outcome data. METHODS: We included healthy parturients at a cervical dilation ≥2 cm, with or without a request for epidural analgesia (n=32 per group). Ultrasound studies of the uterine arteries, umbilical artery and fetal middle cerebral artery were performed before insertion of the epidural catheter, and 30, 60 and 90 min after; the same time-points were assessed in the non-epidural control group. Maternal blood pressure was measured by a continuous non-invasive arterial pressure monitor. RESULTS: Ultrasound studies detected no significant differences in pulsatility indices over time in any blood vessel. In contrast to the control group, maternal blood pressures were significantly lower for all measures after the onset of analgesia compared with baseline values (mean systolic pressure decreased from 132.7 ±â€¯15.9 mmHg to 123.1 ±â€¯14.4 mmHg at 30 min, P=0.003). The mean pH value of the umbilical arterial blood was 7.29 (±0.06) in the epidural group versus 7.31 (±0.08) in the control group (P=0.33). The median Apgar score at 5 min was 10 in both groups. CONCLUSIONS: Pre-placental, fetoplacental and fetal hemodynamics remained stable despite a statistically significant decrease in maternal blood pressure in laboring parturients receiving epidural analgesia.

A Quick Route to Multiple Highly Potent SARS-CoV-2 Main Protease Inhibitors*.

The COVID-19 pathogen, SARS-CoV-2, requires its main protease (SC2MPro ) to digest two of its translated long polypeptides to form a number of mature proteins that are essential for viral replication and pathogenesis. Inhibition of this vital proteolytic process is effective in preventing the virus from replicating in infected cells and therefore provides a potential COVID-19 treatment option. Guided by previous medicinal chemistry studies about SARS-CoV-1 main protease (SC1MPro ), we have designed and synthesized a series of SC2MPro inhibitors that contain ß-(S-2-oxopyrrolidin-3-yl)-alaninal (Opal) for the formation of a reversible covalent bond with the SC2MPro active-site cysteine C145. All inhibitors display high potency with Ki values at or below 100 nM. The most potent compound, MPI3, has as a Ki value of 8.3 nM. Crystallographic analyses of SC2MPro bound to seven inhibitors indicated both formation of a covalent bond with C145 and structural rearrangement from the apoenzyme to accommodate the inhibitors. Virus inhibition assays revealed that several inhibitors have high potency in inhibiting the SARS-CoV-2-induced cytopathogenic effect in both Vero E6 and A549/ACE2 cells. Two inhibitors, MPI5 and MPI8, completely prevented the SARS-CoV-2-induced cytopathogenic effect in Vero E6 cells at 2.5-5 µM and A549/ACE2 cells at 0.16-0.31 µM. Their virus inhibition potency is much higher than that of some existing molecules that are under preclinical and clinical investigations for the treatment of COVID-19. Our study indicates that there is a large chemical space that needs to be explored for the development of SC2MPro inhibitors with ultra-high antiviral potency.

A Speedy Route to Multiple Highly Potent SARS-CoV-2 Main Protease Inhibitors.

The COVID-19 pathogen, SARS-CoV-2, requires its main protease (SC2M Pro ) to digest two of its translated polypeptides to form a number of mature proteins that are essential for viral replication and pathogenesis. Inhibition of this vital proteolytic process is effective in preventing the virus from replication in infected cells and therefore provides a potential COVID-19 treatment option. Guided by previous medicinal chemistry studies about SARS-CoV-1 main protease (SC1M Pro ), we have designed and synthesized a series of SC2M Pro inhibitors that contain ß-( S -2-oxopyrrolidin-3-yl)-alaninal (Opal) for the formation of a reversible covalent bond with the SC2M Pro active site cysteine C145. All inhibitors display high potency with IC 50 values at or below 100 nM. The most potent compound MPI3 has as an IC 50 value as 8.5 nM. Crystallographic analyses of SC2M Pro bound to 7 inhibitors indicated both formation of a covalent bond with C145 and structural rearrangement from the apoenzyme to accommodate the inhibitors. Virus inhibition assays revealed that several inhibitors have high potency in inhibiting the SARS-CoV-2-induced cytopathogenic effect in both Vero E6 and A549 cells. Two inhibitors MP5 and MPI8 completely prevented the SARS-CoV-2-induced cytopathogenic effect in Vero E6 cells at 2.5-5 µM and A549 cells at 0.16-0.31 µM. Their virus inhibition potency is much higher than some existing molecules that are under preclinical and clinical investigations for the treatment of COVID-19. Our study indicates that there is a large chemical space that needs to be explored for the development of SC2M Pro inhibitors with extreme potency. Due to the urgent matter of the COVID-19 pandemic, MPI5 and MPI8 may be quickly advanced to preclinical and clinical tests for COVID-19.

Bepridil is potent against SARS-CoV-2 In Vitro.

Guided by a computational docking analysis, about 30 FDA/EMA-approved small molecule medicines were characterized on their inhibition of the SARS-CoV-2 main protease (MPro). Of these tested small molecule medicines, six displayed an IC50 value in inhibiting MPro below 100 µM. Three medicines pimozide, ebastine, and bepridil are basic small molecules. Their uses in COVID-19 patients potentiate dual functions by both raising endosomal pH to slow SARS-CoV-2 entry into the human cell host and inhibiting MPro in infected cells. A live virus-based microneutralization assay showed that bepridil inhibited cytopathogenic effect induced by SARS-CoV-2 in Vero E6 cells completely at and dose-dependently below 5 µM and in A549 cells completely at and dose-dependently below 6.25 µM. Therefore, the current study urges serious considerations of using bepridil in COVID-19 clinical tests.

Targeting the SARS-CoV-2 Main Protease to Repurpose Drugs for COVID-19

Guided by a computational docking analysis, about 30 FDA/EMA-approved small molecule medicines were characterized on their inhibition of the SARS-CoV-2 main protease (MPro). Of these tested small molecule medicines, six displayed an IC50 value in inhibiting MPro below 100 M. Three medicines pimozide, ebastine, and bepridil are basic small molecules. Their uses in COVID-19 patients potentiate dual functions by both raising endosomal pH to slow SARS-CoV-2 entry into the human cell host and inhibiting MPro in infected cells. A live virus-based microneutralization assay showed that bepridil inhibited cytopathogenic effect induced by SARS-CoV-2 in Vero E6 cells completely at and dose-dependently below 5 M and in A549 cells completely at and dose-dependently below 6.25 M. Therefore, the current study urges serious considerations of using bepridil in COVID-19 clinical tests.

Expressed Protein Ligation without Intein.

Proteins with a functionalized C-terminus such as a C-terminal thioester are key to the synthesis of larger proteins via expressed protein ligation. They are usually made by recombinant fusion to intein. Although powerful, the intein fusion approach suffers from premature hydrolysis and low compatibility with denatured conditions. To totally bypass the involvement of an enzyme for expressed protein ligation, here we showed that a cysteine in a recombinant protein was chemically activated by a small molecule cyanylating reagent at its N-side amide for undergoing nucleophilic acyl substitution with amines including a number of l- and d-amino acids and hydrazine. The afforded protein hydrazides could be used further for expressed protein ligation. We demonstrated the versatility of this activated cysteine-directed protein ligation (ACPL) approach with the successful synthesis of ubiquitin conjugates, ubiquitin-like protein conjugates, histone H2A with a C-terminal posttranslational modification, RNase H that actively hydrolyzed RNA, and exenatide that is a commercial therapeutic peptide. The technique, which is exceedingly simple but highly useful, expands to a great extent the synthetic capacity of protein chemistry and will therefore make a large avenue of new research possible.

Patterns of morphological integration in the dental arches of individuals with malocclusion.

OBJECTIVES: In humans, there is a large range of variation in the form of the maxillary and mandibular dental arches. This variation can manifest as either prognathism or retrognathism in either or both arches, which can cause malocclusion and lead to abnormal masticatory function. This study aims to identify aspects of variation and morphological integration existing in the dental arches of individuals with different types of malocclusion. METHODS: Coordinate landmark data were collected along the gingival margins of 397 scanned dental casts and then analyzed using geometric morphometric techniques to explore arch form variation and patterns of morphological integration within each malocclusion type. RESULTS: Significant differences were identified between Class II forms (increased projection of upper arch relative to the lower arch) and Class III forms (lower arch projection beyond the upper arch) in symmetrical shape variation, including anteroposterior arch discrepancies and abnormal anterior arch divergence or convergence. Partial least squares analysis demonstrated that Class III dental arches have higher levels of covariance between upper and lower arches (RV = 0.91) compared to the dental arches of Class II (RV = 0.78) and Class I (RV = 0.73). These high levels of covariance, however, are on the lower end of the overall range of possible masticatory blocks, indicating weaker than expected levels of integration. CONCLUSIONS: This study provides evidence for patterns of variation in dental arch shape found in individuals with Class II and Class III malocclusions. Moreover, differences in integration found between malocclusion types have ramifications for how such conditions should be studied and treated. Am. J. Hum. Biol. 28:879-889, 2016. © 2016Wiley Periodicals, Inc.

Tuning Interfacial Electron Transfer in Nanostructured Cuprous Oxide Photoelectrochemical Cells with Charge-Selective Molecular Coatings.

The coating of nanostructured films of cuprous oxide with electroactive molecules strongly affects their photoelectrochemical performance in nonaqueous photocells, with photocurrent density increased up to an order of magnitude relative to bare cuprous oxide films or almost completely suppressed, depending on the choice of molecular adsorbant. Among adsorbants that enhance photocurrent, a strong variance of photoelectrochemical behavior is observed with changes in the molecular structure of the sensitizer, associated with differences in the reorganization energy and molecular size, which are interpreted to enhance forward electron transport and impede electrolyte/photocathode recombination, respectively. These results demonstrate that nanostructured cuprous oxide is a promising cathode material for p-type dye-sensitized solar cells.

Phenotypic diversity in white adults with moderate to severe Class II malocclusion.

INTRODUCTION: Class II malocclusion affects about 15% of the population in the United States and is characterized by a convex profile and occlusal disharmonies. The specific etiologic mechanisms resulting in the range of Class II dentoskeletal combinations observed are not yet understood. Most studies describing Class II phenotypic diversity have used moderate sample sizes or focused on younger patients who later in life might outgrow their Class II discrepancies; such a focus might also preclude the visualization of adult Class II features. The majority have used simple correlation methods resulting in phenotypes that might not be generalizable to different samples and thus might not be suitable for studies of malocclusion etiology. The purpose of this study was to address these knowledge gaps by capturing the maximum phenotypic variations in a large sample of white Class II subjects selected with strict eligibility criteria and rigorously standardized multivariate reduction analyses. METHODS: Sixty-three lateral cephalometric variables were measured from the pretreatment records of 309 white Class II adults (82 male, 227 female; ages, 16-60 years). Principal component analysis and cluster analysis were used to generate comprehensive phenotypes to identify the most homogeneous groups of subjects, reducing heterogeneity and improving the power of future malocclusion etiology studies. RESULTS: Principal component analysis resulted in 7 principal components that accounted for 81% of the variation. The first 3 components represented variation on mandibular rotation, maxillary incisor angulation, and mandibular length. The cluster analysis identified 5 distinct Class II phenotypes. CONCLUSIONS: A comprehensive spectrum of Class II phenotypic definitions was obtained that can be generalized to other samples to advance our efforts for identifying the etiologic factors underlying Class II malocclusion.

Phenotypic diversity in white adults with moderate to severe Class III malocclusion.

INTRODUCTION: Class III malocclusion is characterized by a composite of dentoskeletal patterns that lead to the forward positioning of the mandibular teeth in relation to the maxillary teeth and a concave profile. Environmental and genetic factors are associated with this condition, which affects 1% of the population in the United States and imposes significant esthetic and functional burdens on affected persons. The purpose of this study was to capture the phenotypic variation in a large sample of white adults with Class III malocclusion using multivariate reduction methods. METHODS: Sixty-three lateral cephalometric variables were measured from the pretreatment records of 292 white subjects with Class II malocclusion (126 male, 166 female; ages, 16-57 years). Principal component analysis and cluster analysis were used to capture the phenotypic variation and identify the most homogeneous groups of subjects to reduce genetic heterogeneity. RESULTS: Principal component analysis resulted in 6 principal components that accounted for 81.2% of the variation. The first 3 components represented variation in mandibular horizontal and vertical positions, maxillary horizontal position, and mandibular incisor angulation. The cluster model identified 5 distinct subphenotypes of Class III malocclusion. CONCLUSIONS: A spectrum of phenotypic definitions was obtained replicating results of previous studies and supporting the validity of these phenotypic measures in future research of the genetic and environmental etiologies of Class III malocclusion.

Patient preferences regarding 1-visit versus 2-visit root canal therapy.

INTRODUCTION: Patient preferences should be taken into account by clinicians when treatment planning. The purposes of this study were to describe the number of visits patients preferred when undergoing root canal therapy (RCT) and to assess whether their preferences were related to hypothetical treatment success rates. METHODS: Self-administered questionnaires were mailed to 351 consecutive patients scheduled for initial RCT appointments in the University of Iowa College of Dentistry's graduate or faculty endodontic clinic. The questionnaires ascertained demographic information; preferences for 1-visit versus 2-visit RCT given different hypothetical success rate scenarios for the 2 approaches, as well as patient dental history. Univariate frequency distributions were generated, and relationships between hypothetical success rates and patient preferences were evaluated. RESULTS: Questionnaires were returned by 124 patients (35% response). Given equal success rates, 78% of respondents preferred 1-visit RCT, compared with 7% who preferred 2-visit RCT and 16% who would follow their dentist's recommendation. As success rates for 2-visit RCT went from equal to 5% better to 10% better to 20% better compared with 1-visit RCT, the proportion of respondents who preferred 2-visit RCT increased from 7% to 34% to 46% to 65%, respectively. Regardless of success rates, approximately 5% of respondents said they would prefer 2-visit RCT, and 20% would do whatever their dentist recommended. CONCLUSIONS: Although most respondents preferred 1-visit RCT regardless of success rates, many would prefer 2-visit RCT if its success rate were greater than that of 1-visit RCT. This finding confirms the importance of discussing success rates and considering patients' wishes when treatment planning.