Host genetic variation guides hepacivirus clearance, chronicity, and liver fibrosis in mice.

BACKGROUND AIMS: Human genetic variation is thought to guide the outcome of HCV infection, but model systems within which to dissect these host genetic mechanisms are limited. Norway rat hepacivirus, closely related to HCV, causes chronic liver infection in rats but causes acute self-limiting hepatitis in typical strains of laboratory mice, which resolves in 2 weeks. The Collaborative Cross (CC) is a robust mouse genetics resource comprised of a panel of recombinant inbred strains, which model the complexity of the human genome and provide a system within which to understand diseases driven by complex allelic variation. APPROACH RESULTS: We infected a panel of CC strains with Norway rat hepacivirus and identified several that failed to clear the virus after 4 weeks. Strains displayed an array of virologic phenotypes ranging from delayed clearance (CC046) to chronicity (CC071, CC080) with viremia for at least 10 months. Body weight loss, hepatocyte infection frequency, viral evolution, T-cell recruitment to the liver, liver inflammation, and the capacity to develop liver fibrosis varied among infected CC strains. CONCLUSIONS: These models recapitulate many aspects of HCV infection in humans and demonstrate that host genetic variation affects a multitude of viruses and host phenotypes. These models can be used to better understand the molecular mechanisms that drive hepacivirus clearance and chronicity, the virus and host interactions that promote chronic disease manifestations like liver fibrosis, therapeutic and vaccine performance, and how these factors are affected by host genetic variation.

Evaluation of microbial air quality and aerosol distribution in a large dental clinic.

PURPOSE: To evaluate the microbial air quality during dental clinical procedures in a large clinical setting with increasing patient capacity. METHODS: This was a single-center, observational study design evaluating the microbial air quality and aerosol distribution during normal clinical sessions at 5% (sessions 1 and 2) and at > 50% (session 3) treatment capacity of dental aerosol generating procedures. Sessions 1 and 2 were evaluated on the same day with a 30-minute fallow time between the sessions. Session 3 was evaluated on a separate day. For each session, passive air-sampling technique was performed for three collection periods: baseline, treatment, and post-treatment. Blood agar plates were collected and incubated at 37°C for 48 hours. Colonies were counted using an automatic colony counter. Mean colony forming units (CFU) per plate were converted to CFU/m²/h. RESULTS: Kruskal Wallis test was performed to compare the mean CFU/m²/h between the clinic sessions. Statistically significant differences were observed between sessions 1 and 2 (P< 0.05), but not between sessions 2 and 3 (P> 0.05). Combining all clinical sessions, the mean CFU/m²/h were 977 (baseline), 873 (treatment), and 1,631 (post-treatment) for the collection periods. A decrease-to-increase CFU/m²/h trend was observed from baseline to treatment, and from treatment to post-treatment that was observed for all clinic sessions and was irrespective to treatment capacity. Higher amounts of CFU/m²/h were found near the air exhaust outlets for all three clinic sessions. Microbial aerosol distribution is most likely due to the positions and power levels of the air inlets and outlets, and to a lesser extent with patient treatment capacity. CLINICAL SIGNIFICANCE: Dental clinics should be designed and optimized to minimize the risk of airborne transmissions. The results of this study emphasize the need to evaluate dental clinic ventilation systems.

Palatal keratosis associated with reverse (or "backwards") smoking (PKARS).

Palatal mucosal changes secondary to smoking habit have been recognised since at least the 19th century. Smoker's palate, or nicotinic stomatitis is associated with habitual cigar or pipe smoking, regular consumption of thermally hot beverages and more recently has been identified in those who vape. It is considered of negligible malignant transformation risk, typically resolving on habit cessation. However a variant, palatal keratosis associated with reverse smoking (PKARs), is recognised as an established oral potentially malignant disorder. Reverse smoking involves smoking the lit end of the cigarette intraorally and the condition may result in increased incidence of palatal squamous cell carcinoma development. We highlight a case to create awareness of both this unusual smoking habit, and the palate as an overlooked site in oral cancer screening.

The Outcome of a New Teledentistry Initiative in Response to the COVID-19 Pandemic: A Cross-sectional Study.

AIM: The purpose of this study was to assess the overall satisfaction of a new learning experience and evaluate the outcome of LLUSD's educational teledentistry initiative through a survey based on Kirkpatrick's multidimensional model of training evaluation. MATERIALS AND METHODS: An IRB application was approved (#5210385) for a cross-sectional study that included Loma Linda University School of Dentistry (LLUSD) dental students of the class of 2022 and 2023. The 9-question survey consisted of three sections. The first section included demographic questions on gender and the graduating class. The second section was related to perceived teaching effectiveness, attitude, behavior, and significance. The third section included an open-ended question. The survey was distributed by three student investigators. Descriptive statistics were calculated, and categorical variables were compared using the Chi-squared test (χ2 test). RESULTS: The perceived teaching effectiveness of the newly implemented educational initiative was high for increasing the ability to communicate with patients and for screening and identifying the need for referrals. A majority of students believed that teledentistry is an important means to improve patients' access to dental care and that the School has been providing a good educational environment in providing teledentistry sessions to patients. There were no significant differences in the frequencies of positive and negative responses to all questions (N = 6) by gender and by class (p >0.05, in all instances). CONCLUSIONS: Teledentistry eVisits allowed the continuation of patient contact and initial assimilation of patient information. There is potential for this educational initiative to be more actively and comprehensively implemented in the future. CLINICAL SIGNIFICANCE: New educational initiatives allow the continuation of patient contact that will ensure that students will graduate as competent oral health care providers despite challenges imposed by the pandemic.

Impact of COVID-19 on Teaching the Tooth Morphology Course to the New Generation of Learners: A Cross-sectional Study.

AIM: The purpose of the study was two-fold. First, to evaluate students' learning style and relate it to their academic performance. Second, to highlight changes implemented in the tooth morphology (TOMO) course as a response to the coronavirus disease-2019 (COVID-19) pandemic. MATERIALS AND METHODS: The study was performed during 2021-2022 with 101 dental students. Didactic lectures were delivered online and students challenged with nine quizzes and one final examination. Didactic score was calculated by averaging the scores of quizzes and the final exam. Lab score was a combination of five lab projects and the final competency. At course completion, students received a survey on their learning style and how they would like to receive feedback. Kruskal-Wallis test was used to assess differences in didactic and lab scores among groups. RESULTS: Many students perceived themselves as visual learners (39%) followed by kinesthetic (24%), aural (19%), and reader (18%). There was no difference among learning style groups in performance of didactic (p = 0.340) and lab scores (p = 0.845). Students preferred that the instructor talks them through the questions for feedback on quizzes (41%) while they preferred demonstrations when receiving feedback on their wax-ups (51%). Most students (75%) preferred a TOMO teacher that uses demonstrations. 2020-2021 marked the year of the pandemic where all lectures were delivered online and waxing projects were performed at-home. A postpandemic transformation occurred during 2021-2022, reverting to conventional in-person lab sessions while keeping online didactic lectures. CONCLUSION: We conclude that TOMO should be delivered by using various teaching styles rather than focusing on a single method while providing more demonstrations. CLINICAL SIGNIFICANCE: Teaching tooth morphology to the new generation type of learners efficiently will affect the clinical work of dental graduates.

Therapeutic treatment with an oral prodrug of the remdesivir parental nucleoside is protective against SARS-CoV-2 pathogenesis in mice.

The coronavirus disease 2019 (COVID-19) pandemic remains uncontrolled despite the rapid rollout of safe and effective severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines, underscoring the need to develop highly effective antivirals. In the setting of waning immunity from infection and vaccination, breakthrough infections are becoming increasingly common and treatment options remain limited. In addition, the emergence of SARS-CoV-2 variants of concern, with their potential to escape neutralization by therapeutic monoclonal antibodies, emphasizes the need to develop second-generation oral antivirals targeting highly conserved viral proteins that can be rapidly deployed to outpatients. Here, we demonstrate the in vitro antiviral activity and in vivo therapeutic efficacy of GS-621763, an orally bioavailable prodrug of GS-441524, the parent nucleoside of remdesivir, which targets the highly conserved virus RNA-dependent RNA polymerase. GS-621763 exhibited antiviral activity against SARS-CoV-2 in lung cell lines and two different human primary lung cell culture systems. GS-621763 was also potently antiviral against a genetically unrelated emerging coronavirus, Middle East respiratory syndrome CoV (MERS-CoV). The dose-proportional pharmacokinetic profile observed after oral administration of GS-621763 translated to dose-dependent antiviral activity in mice infected with SARS-CoV-2. Therapeutic GS-621763 administration reduced viral load and lung pathology; treatment also improved pulmonary function in COVID-19 mouse model. A direct comparison of GS-621763 with molnupiravir, an oral nucleoside analog antiviral that has recently received EUA approval, proved both drugs to be similarly efficacious in mice. These data support the exploration of GS-441524 oral prodrugs for the treatment of COVID-19.

Infectious Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Virus in Symptomatic Coronavirus Disease 2019 (COVID-19) Outpatients: Host, Disease, and Viral Correlates.

BACKGROUND: Although severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infectious virus isolation in outpatients with coronavirus disease 2019 (COVID-19) has been associated with viral RNA levels and symptom duration, little is known about the host, disease, and viral determinants of infectious virus detection. METHODS: COVID-19 adult outpatients were enrolled within 7 days of symptom onset. Clinical symptoms were recorded via patient diary. Nasopharyngeal swabs were collected to quantitate SARS-CoV-2 RNA by reverse transcriptase polymerase chain reaction and for infectious virus isolation in Vero E6-cells. SARS-CoV-2 antibodies were measured in serum using a validated ELISA assay. RESULTS: Among 204 participants with mild-to-moderate symptomatic COVID-19, the median nasopharyngeal viral RNA was 6.5 (interquartile range [IQR] 4.7-7.6 log10 copies/mL), and 26% had detectable SARS-CoV-2 antibodies (immunoglobulin (Ig)A, IgM, IgG, and/or total Ig) at baseline. Infectious virus was recovered in 7% of participants with SARS-CoV-2 antibodies compared to 58% of participants without antibodies (prevalence ratio [PR] = 0.12, 95% confidence interval [CI]: .04, .36; P = .00016). Infectious virus isolation was also associated with higher levels of viral RNA (mean RNA difference +2.6 log10, 95% CI: 2.2, 3.0; P < .0001) and fewer days since symptom onset (PR = 0.79, 95% CI: .71, .88 per day; P < .0001). CONCLUSIONS: The presence of SARS-CoV-2 antibodies is strongly associated with clearance of infectious virus. Seropositivity and viral RNA levels are likely more reliable markers of infectious virus clearance than subjective measure of COVID-19 symptom duration. Virus-targeted treatment and prevention strategies should be administered as early as possible and ideally before seroconversion. CLINICAL TRIALS REGISTRATION: NCT04405570.

Therapeutic efficacy of an oral nucleoside analog of remdesivir against SARS-CoV-2 pathogenesis in mice.

The COVID-19 pandemic remains uncontrolled despite the rapid rollout of safe and effective SARS-CoV-2 vaccines, underscoring the need to develop highly effective antivirals. In the setting of waning immunity from infection and vaccination, breakthrough infections are becoming increasingly common and treatment options remain limited. Additionally, the emergence of SARS-CoV-2 variants of concern with their potential to escape therapeutic monoclonal antibodies emphasizes the need to develop second-generation oral antivirals targeting highly conserved viral proteins that can be rapidly deployed to outpatients. Here, we demonstrate the in vitro antiviral activity and in vivo therapeutic efficacy of GS-621763, an orally bioavailable prodrug of GS-441524, the parental nucleoside of remdesivir, which targets the highly conserved RNA-dependent RNA polymerase. GS-621763 exhibited significant antiviral activity in lung cell lines and two different human primary lung cell culture systems. The dose-proportional pharmacokinetic profile observed after oral administration of GS-621763 translated to dose-dependent antiviral activity in mice infected with SARS-CoV-2. Therapeutic GS-621763 significantly reduced viral load, lung pathology, and improved pulmonary function in COVID-19 mouse model. A direct comparison of GS-621763 with molnupiravir, an oral nucleoside analog antiviral currently in human clinical trial, proved both drugs to be similarly efficacious. These data demonstrate that therapy with oral prodrugs of remdesivir can significantly improve outcomes in SARS-CoV-2 infected mice. Thus, GS-621763 supports the exploration of GS-441524 oral prodrugs for the treatment of COVID-19 in humans.

Infectious SARS-CoV-2 Virus in Symptomatic COVID-19 Outpatients: Host, Disease, and Viral Correlates.

BACKGROUND: While SARS-CoV-2 infectious virus isolation in outpatients with COVID-19 has been associated with viral RNA levels and symptom duration, little is known about the host, disease and viral determinants of infectious virus detection. METHODS: COVID-19 adult outpatients were enrolled within 7 days of symptom onset. Clinical symptoms were recorded via patient diary. Nasopharyngeal swabs were collected to quantitate SARS-CoV-2 RNA by reverse transcriptase polymerase chain reaction and for infectious virus isolation in Vero E6-cells. SARS-CoV-2 antibodies were measured in serum using a validated ELISA assay. RESULTS: Among 204 participants with mild-to-moderate symptomatic COVID19, the median nasopharyngeal viral RNA was 6.5 (IQR 4.7-7.6 log10 copies/mL), and 26% had detectable SARS-CoV-2 antibodies (IgA, IgM, IgG, and/or total Ig) at baseline. Infectious virus was recovered in 7% of participants with SARS-CoV-2 antibodies compared to 58% of participants without antibodies (probability ratio (PR)=0.12, 95% CI: 0.04, 0.36; p=0.00016). Infectious virus isolation was also associated with higher levels of viral RNA (mean RNA difference +2.6 log10, 95% CI: 2.2, 3.0; p<0.0001) and fewer days since symptom onset (PR=0.79, 95% CI: 0.71, 0.88 per day; p<0.0001). CONCLUSIONS: The presence of SARS-CoV-2 antibodies is strongly associated with clearance of infectious virus isolation. Seropositivity and viral RNA levels are likely more reliable markers of infectious virus clearance than subjective measure of COVID-19 symptom duration. Virus-targeted treatment and prevention strategies should be administered as early as possible and ideally before seroconversion. CLINICALTRIALSGOV IDENTIFIER: NCT04405570.

A clinical investigation of dental evacuation systems in reducing aerosols.

BACKGROUND: The route of transmission of severe acute respiratory syndrome coronavirus 2 has challenged dentistry to improve the safety for patients and the dental team during various treatment procedures. The purpose of this study was to evaluate and compare the effectiveness of dental evacuation systems in reducing aerosols during oral prophylactic procedures in a large clinical setting. METHODS: This was a single-center, controlled clinical trial using a split-mouth design. A total of 93 student participants were recruited according to the inclusion and exclusion criteria. Aerosol samples were collected on blood agar plates that were placed around the clinic at 4 treatment periods: baseline, high-volume evacuation (HVE), combination (HVE and intraoral suction device), and posttreatment. Student operators were randomized to perform oral prophylaxis using ultrasonic scalers on 1 side of the mouth, using only HVE suction for the HVE treatment period and then with the addition of an intraoral suction device for the combination treatment period. Agar plates were collected after each period and incubated at 37 °C for 48 hours. Colony-forming unit (CFU) counts were determined using an automatic colony counter. RESULTS: The use of a combination of devices resulted in significant reductions in CFUs compared with the use of the intraoral suction device alone (P < .001). The highest amounts of CFUs were found in the operating zone and on patients during both HVE and combination treatment periods. CONCLUSIONS: Within limitations of this study, the authors found significant reductions in the amount of microbial aerosols when both HVE and an intraoral suction device were used. PRACTICAL IMPLICATIONS: The combination of HVE and intraoral suction devices significantly decreases microbial aerosols during oral prophylaxis procedures.

A multicenter study of dental curricula in Asia/Pacific nations: The views and experiences of final-year dental students.

OBJECTIVES: The objective of this qualitative study was to gain greater understanding of final-year dental students' views on and experience of their dental curricula in 4 universities from different Asia/Pacific countries, including New Zealand, Australia, and Hong Kong (China). METHODS: A qualitative study approach was used, with semistructured interviews conducted with final-year students from each of the 4 universities. RESULTS: Interviews were conducted with 60 final-year dental students, and 5 main themes were extracted from the interviews: (1) the definition of an "ideal" dental curriculum, (2) theoretical teaching, (3) transitional tools, (4) assessment, and (5) grading. CONCLUSION: The findings provide insight into final-year students' views of dental curricula and suggestions on possible areas of reform in the dental curriculum. Further investigations are necessary to provide a curriculum that enables students to become competent, future-ready dental practitioners.

Comparative therapeutic efficacy of remdesivir and combination lopinavir, ritonavir, and interferon beta against MERS-CoV.

Middle East respiratory syndrome coronavirus (MERS-CoV) is the causative agent of a severe respiratory disease associated with more than 2468 human infections and over 851 deaths in 27 countries since 2012. There are no approved treatments for MERS-CoV infection although a combination of lopinavir, ritonavir and interferon beta (LPV/RTV-IFNb) is currently being evaluated in humans in the Kingdom of Saudi Arabia. Here, we show that remdesivir (RDV) and IFNb have superior antiviral activity to LPV and RTV in vitro. In mice, both prophylactic and therapeutic RDV improve pulmonary function and reduce lung viral loads and severe lung pathology. In contrast, prophylactic LPV/RTV-IFNb slightly reduces viral loads without impacting other disease parameters. Therapeutic LPV/RTV-IFNb improves pulmonary function but does not reduce virus replication or severe lung pathology. Thus, we provide in vivo evidence of the potential for RDV to treat MERS-CoV infections.

Broad spectrum antiviral remdesivir inhibits human endemic and zoonotic deltacoronaviruses with a highly divergent RNA dependent RNA polymerase.

The genetically diverse Orthocoronavirinae (CoV) family is prone to cross species transmission and disease emergence in both humans and livestock. Viruses similar to known epidemic strains circulating in wild and domestic animals further increase the probability of emergence in the future. Currently, there are no approved therapeutics for any human CoV presenting a clear unmet medical need. Remdesivir (RDV, GS-5734) is a monophosphoramidate prodrug of an adenosine analog with potent activity against an array of RNA virus families including Filoviridae, Paramyxoviridae, Pneumoviridae, and Orthocoronavirinae, through the targeting of the viral RNA dependent RNA polymerase (RdRp). We developed multiple assays to further define the breadth of RDV antiviral activity against the CoV family. Here, we show potent antiviral activity of RDV against endemic human CoVs OC43 (HCoV-OC43) and 229E (HCoV-229E) with submicromolar EC50 values. Of known CoVs, the members of the deltacoronavirus genus have the most divergent RdRp as compared to SARS- and MERS-CoV and both avian and porcine members harbor a native residue in the RdRp that confers resistance in beta-CoVs. Nevertheless, RDV is highly efficacious against porcine deltacoronavirus (PDCoV). These data further extend the known breadth and antiviral activity of RDV to include both contemporary human and highly divergent zoonotic CoV and potentially enhance our ability to fight future emerging CoV.

Assessment of the Anterior Loop of the Mental Nerve Using Cone Beam Computerized Tomography Scan.

The purpose of this study is to use cone-beam computerized tomography (CBCT) scans with oblique-transverse reconstruction modality to measure and compare the anterior loop length (AnLL) of the mental nerve between gender and age groups and to compare the difference between the right and left sides. Sixty-one female and 61 male CBCT scans were randomly selected for each age group: 21-40, 41-60, and 61-80 years. Both right- and left-side AnLLs were measured in each subject using i-CATVision software to measure AnLLs on the oblique transverse plane using multiplanar reconstruction. The anterior loop was identified in 85.2% of cases, with the mean AnLL of the 366 subjects (732 hemimandibles) being 1.46 ± 1.25 mm with no statistically significant difference between right and left sides or between different gender groups. However, the mean AnLL in the 21-40 year group (1.89 ± 1.35 mm) was larger than the AnLL in the 41-60 year group (1.35 ± 1.19 mm) and the 61-80 year group (1.13 ± 1.08 mm). In conclusion, when placing implants in close proximity to mental foramina, caution is recommended to avoid injury to the inferior alveolar nerve. No fixed distance anteriorly from the mental foramen should be considered safe. Using CBCT scans with the oblique-transverse method to accurately identify and measure the AnLL is of utmost importance in avoiding and protecting its integrity.

Gingival displacement for impression making in fixed prosthodontics: contemporary principles, materials, and techniques.

The clinical success and longevity of indirect restorations depend on the careful and accurate completion of several procedures. One of the challenging procedures is management of the gingival tissues and gingival esthetics. The goal for management of gingival tissues and gingival esthetics is to maintain the normal appearance of healthy gingival. Achieving this goal requires optimal health before treatment and minimal trauma during treatment. The best way of optimizing health and minimizing trauma is to avoid contacting the gingiva with restorative materials.

The effect of angulation sensors on implant placement.

It is hypothesized that use of an angulation sensor could increase the alignment accuracy of multiple implant osteotomies without requiring the use of guided-surgical templates. Therefore, the purpose of this study was to determine if use of such a sensor mounted on a surgical handpiece could improve implant placement.

Additively enhanced antiproliferative effect of interferon combined with proanthocyanidin on bladder cancer cells.

Although interferon (IFN) has been often used as immunotherapy for bladder cancer, its efficacy is rather unsatisfactory, demanding further improvement. Combination therapy is one of viable options, and grape seed proanthocyanidin (GSP) could be such an agent to be used with IFN because it has been shown to have anticancer activity. We thus investigated whether combination of IFN and GSP might enhance the overall antiproliferative effect on bladder cancer cells in vitro. Human bladder cancer T24 cells were employed and treated with the varying concentrations of recombinant IFN-α(2b) (0-100,000 IU/ml), GSP (0-100 µg/ml), or their combinations. IFN-α(2b) alone led to a ~50% growth reduction at 20,000 (20K) IU/ml, which further declined to ~67% at ≥50K IU/ml. Similarly, GSP alone induced a ~35% and ~100% growth reduction at 25 and ≥50 µg/ml, respectively. When IFN-α(2b) and GSP were then combined, combination of 50K IU/ml IFN-α(2b) and 25 µg/ml GSP resulted in a drastic >95% growth reduction. Cell cycle analysis indicated that such an enhanced growth inhibition was accompanied by a G(1) cell cycle arrest. This was further confirmed by Western blot analysis revealing that expressions of G(1)-specific cell cycle regulators (CDK2, CDK4, cyclin E and p27/Kip1) were distinctly modulated with such IFN-α(2b)/GSP treatment. Therefore, these findings support the notion that combination of IFN-α(2b) and GSP is capable of additively enhancing antiproliferative effect on T24 cells with a G(1) cell cycle arrest, implying an adjuvant therapeutic modality for superficial bladder cancer.

Retrieval of a fractured zirconia implant abutment using a modified crown and bridge remover: a clinical report.

This clinical report describes a novel method to retrieve the internal connection of a fractured zirconia abutment through modification of a crown and bridge remover. Furthermore, the strengths and limitations of using a zirconia implant abutment will be highlighted.