Seroprevalence of group B Coxsackieviruses: retrospective study in an Italian population

Purpose Group B Coxsackieviruses (CVB) include 6 serotypes (B1‐6) responsible for a wide range of clinical diseases. Since no recent seroepidemiologic data are available in Italy, the study aim was to investigate CVB seroprevalence in a wide Italian population. Methods The study retrospectively included 2,459 subjects referring to a large academic hospital in Rome (Italy) in the period 2004‐2016. Seroprevalence rates and neutralizing antibodies (nAb) titers were evaluated in relation to years of observation and subjects’ characteristics. Results Positivity for at least one serotype was detected in 69.1% of individuals. Overall, the prevalent serotype was B4, followed by B3 (33.3%), B5 (26.2%), B1 (12.7%), B2 (11.0%), and B6 (1.7%). For B2, a significant decrease in seroprevalence over years was observed. Positivity to at least one virus was 25.2% in children aged 0‐2 years, but significantly increased in pre‐school (3‐5 yr) (50.3%) and school (6‐10 yr) children (70.4%). Higher nAb responses for B3 and B4 were observed in children aged 3‐5 years. Conclusion A high overall CVB prevalence was found. Type‐specific variations in prevalence over time probably reflect the fluctuations in circulation typical of Enteroviruses. Children are at greater risk for CVB infection given the high number of seronegative subjects aged 0‐10 years

Association of Variants in the SPTLC1 Gene With Juvenile Amyotrophic Lateral Sclerosis

Importance: Juvenile amyotrophic lateral sclerosis (ALS) is a rare form of ALS characterized by age of symptom onset less than 25 years and a variable presentation.Objective: To identify the genetic variants associated with juvenile ALS.Design, Setting, and Participants: In this multicenter family-based genetic study, trio whole-exome sequencing was performed to identify the disease-associated gene in a case series of unrelated patients diagnosed with juvenile ALS and severe growth retardation. The patients and their family members were enrolled at academic hospitals and a government research facility between March 1, 2016, and March 13, 2020, and were observed until October 1, 2020. Whole-exome sequencing was also performed in a series of patients with juvenile ALS. A total of 66 patients with juvenile ALS and 6258 adult patients with ALS participated in the study. Patients were selected for the study based on their diagnosis, and all eligible participants were enrolled in the study. None of the participants had a family history of neurological disorders, suggesting de novo variants as the underlying genetic mechanism.Main Outcomes and Measures: De novo variants present only in the index case and not in unaffected family members.Results: Trio whole-exome sequencing was performed in 3 patients diagnosed with juvenile ALS and their parents. An additional 63 patients with juvenile ALS and 6258 adult patients with ALS were subsequently screened for variants in the SPTLC1 gene. De novo variants in SPTLC1 (p.Ala20Ser in 2 patients and p.Ser331Tyr in 1 patient) were identified in 3 unrelated patients diagnosed with juvenile ALS and failure to thrive. A fourth variant (p.Leu39del) was identified in a patient with juvenile ALS where parental DNA was unavailable. Variants in this gene have been previously shown to be associated with autosomal-dominant hereditary sensory autonomic neuropathy, type 1A, by disrupting an essential enzyme complex in the sphingolipid synthesis pathway.Conclusions and Relevance: These data broaden the phenotype associated with SPTLC1 and suggest that patients presenting with juvenile ALS should be screened for variants in this gene.</p

Biofilm in Genital Ecosystem: A Potential Risk Factor for Chlamydia trachomatis Infection

In healthy women, the cervicovaginal microbiota is mostly populated by Lactobacillus spp., the main host defense factor of the female genital tract. In addition to Lactobacilli, other microorganisms populate the cervicovaginal microbiota, like Candida spp. and Gardnerella vaginalis. The overgrowth of Candida spp. or G. vaginalis, known as biofilm-producing microorganisms in the genital ecosystem, may lead to microbial dysbiosis that increases the risk of acquiring sexually transmitted infections, like Chlamydia trachomatis. C. trachomatis, the leading cause of bacterial sexually transmitted diseases, is still considered an important public health problem worldwide because of the impact of asymptomatic infections on long-term reproductive sequelae, including pelvic inflammatory disease and infertility. The aim of our study was to investigate the interaction between C. trachomatis and the biofilm produced by Candida albicans or Gardnerella vaginalis, evaluating whether the biofilm can harbor C. trachomatis and influence its survival as well as its infectious properties. In order to do so, we developed an in vitro coculture transwell-based biofilm model. Our findings proved, for the first time, that C. trachomatis, an intracellular obligate pathogen, survived, for up to 72 hours after exposure, inside the biofilm produced by C. albicans or G. vaginalis, retaining its infectious properties, as evidenced by the typical chlamydial inclusions observed in the cell monolayer (chlamydial inclusion-forming units at 72 h: 9255 ± 1139 and 9873 ± 1015, respectively). In conclusion, our results suggest that the biofilm related to Candida or Gardnerella genital infections may act as a reservoir of C. trachomatis and, thus, contribute to the transmission of the infection in the population as well as to its dissemination into the upper genital tract, increasing the risk of developing severe reproductive sequelae

TLR9 is expressed in human papillomavirus-positive cervical cells and is overexpressed in persistent infections

Control of human papillomavirus (HPV) infection involves the activation of Toll-like receptors (TLRs), key components of the mucosal antiviral response. Available studies on TLR expression in HPV-positive cervical cells are limited and reported conflicting results. This study quantified TLR 2, 3, 4, 7 and 9 transcripts in low-risk (LR) and high-risk (HR) HPV-positive and HPV-negative cervical samples from 154 women attending a gynaecological clinic. Expression levels of TLR 2, 3, 4 and 7 did not differ among samples, whereas TLR9 levels were quite significantly higher in LR and marginally significant in HR HPV-positive samples, with respect to the HPV-negative samples. Interestingly, in a subgroup of women with documented previous HPV-infection, TLR9 levels were extremely higher in patients persistently positive to the same HPV genotype for more than one year, with respect to women who cleared HPV infection and to those re-infected with a different genotype. These findings implicate TLR9 in the response to LR and HR HPVs, including HPV 16 known to interfere with TLR9 transcription in cell lines. Elevated TLR9 levels without HPV clearance in persistently infected women could drive inflammation thereby contributing to cervical cancer risk

Oral hygiene habits and use of fluoride in developmental age: role of parents and impact on their children

Introduction. In healthcare, the need to pay more attention to the achievement of two objectives within the society arises: health promotion and prevention in terms of nutrition, good education, sport, and health education. Scientific evidence shows that adequate health standards must be learned since childhood through the help of parents and appropriate school projects. Parental intervention must be appropriate to support the responsibility of their children’s health. In oral health, it has been established for many years that there is a correlation between parental behaviors and lifestyles and children’s attitude. The aim of this study is to verify the close relation between behaviors, habits, lifestyles, and the knowledge of parents about their oral health and, consequently, their focus and care for their own children’s oral health. Furthermore, the awareness of parents about the importance and use of fluorine was to be determined. Materials and Methods. The study lasted 15 months and was conducted from April 2018 to July 2019: an anonymous 29-question questionnaire was administered to all parents who accompanied their children (aged between 3 and 12 years) going under treatment in the Pediatric Dentistry Unit of the University Hospital Policlinico Umberto I, Rome. Anamnestic data, sociodemographic context (e.g., educational level and occupation), oral health habits, and prevention of parents and children and fluoride knowledge were investigated. The study received ethical approval. 204 questionnaires were collected. The data gathered were recorded with a specifically designed computer program and collected and analyzed using a Microsoft Excel 10 database. Data were evaluated using standard statistical analysis software; descriptive statistics including mean ± SD values and percentage were calculated for each variable. The relationship between the age of parents, between mother or father and the parents’ degree of education levels, and the knowledge for their own children’s oral health was explored using the chi-square test of homogeneity and Fisher’s exact test ( value of &lt; 0.05 considered as statistically significant). Results. From the acquired data, it is possible to deduce that the major respondents were mothers aged from 36 to 45, while only a small part were fathers aged above 45 years. Questions related to parents’ oral hygiene habits were included in the questionnaire, and from the sample taken into consideration, it emerges that 64.7% of the respondents (67.1% mothers and 57.7% fathers) periodically attend a dental office for a checkup, 20.9% tend to postpone the treatment, and 15.2% go there just for emergency. Some of the questions showed that 80% of the interviewed subjects use fluoride toothpaste for their child’s oral hygiene. Conclusion. Prevention in childhood, in addition to being synonymous with monitoring the oral health of the child, means first of all to pay attention to parents who are the main behavioral reference. It emerged that there is no adequate knowledge about fluorine, especially when the subjects have a low educational level. A role of fundamental importance for the diffusion of adequate concepts in the field of oral hygiene is covered, according to the data received from the study carried out, by the dentist and dental hygienist

A simple, fast and reliable scan-based technique as a novel approach to quantify intracellular bacteria

Background: Quantification of intracellular bacteria is fundamental in many areas of cellular and clinical microbiology to study acute and chronic infections. Therefore, rapid, accurate and low-cost methods represent valuable tools in determining bacterial ability to persist and proliferate within eukaryotic cells. Results: Herein, we present the first application of the immunofluorescence In-Cell Western (ICW) assay aimed at quantifying intracellular bacteria in in vitro infection models. The performance of this new approach was evaluated in cell culture infection models using three microorganisms with different lifestyles. Two facultative intracellular bacteria, the fast-growing Shigella flexneri and a persistent strain of Escherichia coli, as well as the obligate intracellular bacterium Chlamydia trachomatis were chosen as bacterial models. The ICW assay was performed in parallel with conventional quantification methods, i.e. colony forming units (CFUs) and inclusion forming units (IFUs). The fluorescence signal intensity values from the ICW assay were highly correlated to CFU/IFUs counting and showed coefficients of determination (R2), ranging from 0,92 to 0,99. Conclusions: The ICW assay offers several advantages including sensitivity, reproducibility, high speed, operatorindependent data acquisition and overtime stability of fluorescence signals. All these features, together with the simplicity in performance, make this assay particularly suitable for high-throughput screening and diagnostic approaches

Association of Variants in the SPTLC1 Gene with Juvenile Amyotrophic Lateral Sclerosis

Importance: Juvenile amyotrophic lateral sclerosis (ALS) is a rare form of ALS characterized by age of symptom onset less than 25 years and a variable presentation. Objective: To identify the genetic variants associated with juvenile ALS. Design, Setting, and Participants: In this multicenter family-based genetic study, trio whole-exome sequencing was performed to identify the disease-associated gene in a case series of unrelated patients diagnosed with juvenile ALS and severe growth retardation. The patients and their family members were enrolled at academic hospitals and a government research facility between March 1, 2016, and March 13, 2020, and were observed until October 1, 2020. Whole-exome sequencing was also performed in a series of patients with juvenile ALS. A total of 66 patients with juvenile ALS and 6258 adult patients with ALS participated in the study. Patients were selected for the study based on their diagnosis, and all eligible participants were enrolled in the study. None of the participants had a family history of neurological disorders, suggesting de novo variants as the underlying genetic mechanism. Main Outcomes and Measures: De novo variants present only in the index case and not in unaffected family members. Results: Trio whole-exome sequencing was performed in 3 patients diagnosed with juvenile ALS and their parents. An additional 63 patients with juvenile ALS and 6258 adult patients with ALS were subsequently screened for variants in the SPTLC1 gene. De novo variants in SPTLC1 (p.Ala20Ser in 2 patients and p.Ser331Tyr in 1 patient) were identified in 3 unrelated patients diagnosed with juvenile ALS and failure to thrive. A fourth variant (p.Leu39del) was identified in a patient with juvenile ALS where parental DNA was unavailable. Variants in this gene have been previously shown to be associated with autosomal-dominant hereditary sensory autonomic neuropathy, type 1A, by disrupting an essential enzyme complex in the sphingolipid synthesis pathway. Conclusions and Relevance: These data broaden the phenotype associated with SPTLC1 and suggest that patients presenting with juvenile ALS should be screened for variants in this gene.