Deep sequencing detects human papillomavirus (HPV) in cervical cancers negative for HPV by PCR

Background Human papillomavirus (HPV) is a necessary cause of cervical cancer, although some invasive cervical cancers may test negative by HPV PCR. We previously requested all invasive cervical cancers in Sweden during 10 years and subjected them to PCR. We also optimised methods for deep sequencing of formalin-fixed paraffin-embedded samples.Methods Using Novaseq 6000, we simultaneously sequenced total DNA and cDNA from 392 HPV PCR-negative cervical cancers. Non-human reads were queried against all known HPVs. The complete database now contains PCR and/or deep sequencing data on 2850 invasive cervical cancers.Results HPV sequences were detected in 169/392 of HPV PCR-negative cervical cancers. Overall, 30 different HPV types were detected, but only 5 types were present in proportions above 3% of cancers. More than 92% of tumours were HPV-positive in PCR and/or sequencing (95% confidence interval: 91.1–93.1%). Exploring possible reasons for failure to previously detect HPV suggest that more sensitive type-specific PCRs for HPV 31, 33, 45 and 73 targeting retained regions of HPV would have detected most of these (117/392).Conclusions Unbiased deep sequencing provides comprehensive data on HPV types in cervical cancers and appears to be an important tool for quality assurance of HPV screening.Subject terms: Infectious-disease diagnostics, Viral infection, Cervical cancer     

Comparative evaluation of pulpal repair after direct pulp capping using stem cell therapy and biodentine: An animal study

The response of the dentin–pulp complex in rat teeth was investigated after direct capping with biodentine with or without bone marrow‐derived stem cells (BMDSCs). Following mechanical exposure, pulps were randomly capped with one of the followings materials: calcium hydroxide, biodentine or 1 × 10⁵ BMDSCs mL^?1 + biodentine. Histological examination was performed by light microscopy after 1, 3 and 5 weeks. Inflammatory reaction, necrotic tissue formation and calcific bridge formation were scored. Analysis showed that compared with the effects of calcium hydroxide or biodentine, BMDSCs + biodentine substantially reduced inflammatory reaction and necrotic tissue while promoting calcified tissue formation. Therefore, the combination of biodentine and BMDSCs could potentially stimulate pulp tissue regeneration after direct pulp capping.     

Synthesis,biological evaluation and in silico molecular docking of novel 1-hydroxy-naphthyl substituted heterocycles

The versatile precursor 2-acetyl-4-allyl-1-hydroxy naphthalene was synthesized efficiently via Claisen rearrangement 2-acetyl-1-allyloxynaphthalene. The Claisen-Schmidt condensation of latter precursor afforded the corresponding chalcones which were exploited to synthesize a series of potential heterocycles such as pyrazoline, isoxazoline, benzocoumarin and benzoflavone. The synthesized products showed potent antioxidant and antimicrobial activities. Chalcone 3c, naphthyl pyrazoline 6b and hydroxycoumarin 13 exhibited the highest activity as antioxidants. Their binding mode showed specialized recognition of hydroxycoumarin 13 with the triad key amino acids at the active site of the oxidoreductase enzyme (PDB code 1DXO). 1-Hydroxynaphth-2-yl pyrazoline (6b) revealed the highest efficacy against both Gram positive and negative bacterial species. In silico molecular docking of pyrazoline 6b endorsed its proper binding at the active site of the 2EX6 enzyme which explains its potent antibacterial activity in comparison with standard ampicillin.     

Two-stage vertical flow multi-soil-layering (MSL) technology for efficient removal of coliforms and human pathogens from domestic wastewater in rural areas under arid climate

This paper investigates the removal efficiency of organic matter, nitrogen, phosphorus, coliforms and pathogens from rural domestic wastewater in a two-stage vertical flow multi-soil-layering (MSL) system. The effects of wastewater quality, season and arid climate conditions on pollutants removal efficiency by the system were examined for one year. The experimental setup included two similar MSL systems composed of two layers: soil-mixture-layers (SML) and gravel permeable layers (PL) that are arranged in a brick like pattern. The applied hydraulic loading rate was 1000 L m^?2 day^?1. Results showed that most of the physicochemical contaminants elimination occurred while the wastewater percolated through the first MSL stage. The second stage demonstrated an improvement in the reduction of all pollutants, especially fecal bacteria indicators and pathogens. The mean overall removal rates performed by the two-stage MSL system were 97% for TSS, 96% for BOD₅, 91% for COD, 96% for TN and 95% for TP. For bacterial indicators, the combination of two-stage MSL system achieved high log removals between 2.21 and 3.15 log units. Contaminants reduction processes in MSL technology are more dependent on internal than external environmental factors. The effectiveness of the two-stage MSL system to treat domestic wastewater was strongly influenced by wastewater quality. Significant relationships between influent contaminants level and their removal efficiency were found. The efficiency of MSL technology to reduce contaminants is not sensitive to season and air temperature fluctuations. This is due to the capacity of MSL system materials to withstand the air temperature variation, which highlights one of the advantages of MSL’s technology. Wastewater quality is the most important factor affecting the removal of contaminants in the MSL, which could be a critical parameter to considered when designing MSL system. Two-stage MSL system achieved a high treated wastewater quality amenable for treated wastewater reuse in agriculture recommended by Moroccan code of practice. Therefore, the combination of two-stage vertical flow MSL system could be considered an efficient and promising domestic wastewater treatment solution in arid countries to promote environmental protection and wastewater reuse.     

The prevalence of oral mucosal lesions in patients visiting a dental school in Southern India.

The purpose of the present study was to evaluate the prevalence of oral mucosal lesions in Manipal, Karnataka State, India. A total of 1190 subjects who visited the department of oral medicine and radiology for diagnosis of various oral complaints over a period of 3 months were interviewed and clinically examined for oral mucosal lesions. The result showed the presence of one or more mucosal lesions in (41.2%) of the population. Fordyce's condition was observed most frequently (6.55%) followed by frictional keratosis (5.79%), fissured tongue (5.71%), leukoedema (3.78%), smoker's palate (2.77%), recurrent aphthae, oral submucous fibrosis (2.01%), oral malignancies (1.76%), leukoplakia (1.59%), median rhomboid glossitis (1.50%), candidiasis (1.3%), lichen planus (1.20%), varices (1.17%), traumatic ulcer and oral hairy leukoplakia (1.008%), denture stomatitis, geographic tongue, betel chewer's mucosa and irritational fibroma (0.84%), herpes labialis, angular cheilitis (0.58%), and mucocele (0.16%). Mucosal lesions like tobacco-related lesions (leukoplakia, smoker's palate, oral submucous fibrosis, and oral malignancies) were more prevalent among men than among women. Denture stomatitis, herpes labialis, and angular cheilitis occurred more frequently in the female population.