Overview of the Clinical and Molecular Features of Legionella Pneumophila: Focus on Novel Surveillance and Diagnostic Strategies.

Legionella pneumophila (L. pneumophila) is one of the most threatening nosocomial pathogens. The implementation of novel and more effective surveillance and diagnostic strategies is mandatory to prevent the occurrence of legionellosis outbreaks in hospital environments. On these bases, the present review is aimed to describe the main clinical and molecular features of L. pneumophila focusing attention on the latest findings on drug resistance mechanisms. In addition, a detailed description of the current guidelines for the disinfection and surveillance of the water systems is also provided. Finally, the diagnostic strategies available for the detection of Legionella spp. were critically reviewed, paying the attention to the description of the culture, serological and molecular methods as well as on the novel high-sensitive nucleic acid amplification systems, such as droplet digital PCR.

Reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) as a user-friendly system to detect SARS-CoV-2 infection: a multicentric study.

Although reverse transcriptase quantitative PCR remains the gold standard to perform viral detection, reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) is already used to perform diagnosis of various infections. This work reports the results of a multicentric study performed in Sicily to evaluate the diagnostic power of an RT-LAMP kit for the diagnosis of SARS-CoV-2 infection on a total of 551 samples collected in January and February 2021, revealing sensitivity, specificity, accuracy, positive and negative predictive values ≥95%. Our results suggest the potential employment of this kit as a screening test to be used where fast and reliable results are demanded without the need for expensive instruments and highly-skilled personnel.

Genetic and Epigenetic Alterations Induced by Pesticide Exposure: Integrated Analysis of Gene Expression, microRNA Expression, and DNA Methylation Datasets.

Environmental or occupational exposure to pesticides is considered one of the main risk factors for the development of various diseases. Behind the development of pesticide-associated pathologies, there are both genetic and epigenetic alterations, where these latter are mainly represented by the alteration in the expression levels of microRNAs and by the change in the methylation status of the DNA. At present, no studies have comprehensively evaluated the genetic and epigenetic alterations induced by pesticides; therefore, the aim of the present study was to identify modifications in gene miRNA expression and DNA methylation useful for the prediction of pesticide exposure. For this purpose, an integrated analysis of gene expression, microRNA expression, and DNA methylation datasets obtained from the GEO DataSets database was performed to identify putative genes, microRNAs, and DNA methylation hotspots associated with pesticide exposure and responsible for the development of different diseases. In addition, DIANA-miRPath, STRING, and GO Panther prediction tools were used to establish the functional role of the putative biomarkers identified. The results obtained demonstrated that pesticides can modulate the expression levels of different genes and induce different epigenetic alterations in the expression levels of miRNAs and in the modulation of DNA methylation status.

A Combined Multidisciplinary Intervention for Health Promotion in the Workplace: A Pilot Study.

The aim of this study was to assess the effects of a joint health promotion intervention on a cohort of healthcare workers (HCWs) who had at least one cardiovascular risk factor. The HCWs were assessed at three different times, i.e., time zero (T0), after 6 months (T6), and after 12 months (T12). The following parameters were measured at a medical examination: physical activity, blood pressure, waist circumference, body mass index (BMI), routine laboratory tests, plicometric analysis, work ability index (WAI), and body image dissatisfaction (BID). Among the 447 HCWs, 38 HCWs were included in the study; 45% (n = 17) were male. At T12, the average blood pressure, waist/hip ratio (WHR) index, BMI, total cholesterol, triglyceride level, and blood glucose values were reduced. The levels of physical activity and adherence to the Mediterranean diet had progressively increased. The WAI showed a significant shift from low to good work performance at T12, as well as BID score. This is the first study that has analyzed work performance in relation to a workplace health promotion through a multidisciplinary approach. This health promotion intervention that combined diet and sport activity has led to a significant change in HCWs' lifestyles and body perceptions, as well as their ability to work. This project highlights the importance of using a multidisciplinary approach and the workplace setting in health promotion programs.

Oxidative stress and DNA damage in agricultural workers after exposure to pesticides.

BACKGROUND: Recent epidemiological studies on workers describe that exposure to pesticides can induce oxidative stress by increased production of free radicals that can accumulate in the cell and damage biological macromolecules, for example, RNA, DNA, DNA repair proteins and other proteins and/or modify antioxidant defense mechanisms, as well as detoxification and scavenger enzymes. This study aimed to assess oxidative stress and DNA damage among workers exposed to pesticides. METHODS: For this purpose, 52 pesticide exposed workers and 52 organic farmers were enrolled. They were assessed: the pesticide exposure, thiobarbituric acid reactive substances (TBARS), total glutathione (TG), oxidized glutathione levels (GSSG), and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), levels. RESULTS: Correlation between pesticide exposure was positively associated with high TBARS and 8-oxodG levels (p <  0.001). A negative association was founded with TG and GSSG and pesticide exposure. CONCLUSIONS: The present investigation results seem to indicate a mild augment in oxidative stress associated with pesticide exposure, followed by an adaptive response to increase the antioxidant defenses to prevent sustained oxidative adverse effects stress.

Vaccine-preventable disease in healthcare workers in Sicily (Italy): seroprevalence against measles.

Aim: Measles is one of the most infectious communicable diseases. The objective of this study was to determine the immunity to measles of healthcare workers (HCWs) operating in three hospitals of Catania. Methods: A total of 549 HCWs underwent measles screening. A 5 ml blood sample was taken from each worker to measure IgG antibody levels. Results: Overall seroprotection was 86%. Unvaccinated HCWs agreed to undergo the vaccination offered by the hospital. Furthermore, it was found that younger workers are less seroprotected than older ones. Conclusion: Model legislation may be helpful to countries wishing to implement immunization requirements in healthcare settings in order to virtually eliminate the risk of acquiring and spreading measles in healthcare settings.

Tuberculosis screening among healthcare workers in Sicily, Italy.

Tuberculosis (TB) is a diffuse communicable disease; and healthcare workers (HCWs) are among the at-risk populations for the disease. This study reports the result of TB surveillance in HCWs operating in three hospitals: one classified as 'low' risk for TB and two as 'medium' risk. A total of 2228 HCWs underwent TB screening using the standard Mantoux technique (also known as the tuberculin sensitivity test [TST]). Interferon-γ release assay (IGRA) was carried out in TST-positive subjects. Results showed an overall positivity of 21% to TST and 2% to IGRA test. TST and IGRA results were statistically associated with HCWs born after 1981. This result is due to university policies that highly recommend TB immunization. Data support the use of IGRA assay in TST-positive HCWs, in order to identify latent TB infection from vaccination.

High HOMA-IR Index in Healthcare Shift Workers.

Background and objectives: Evidence shows that shift work may be correlated with insulin resistance (IR). Therefore its estimation in clinical and prevention practice is of great significance. A cross-sectional study was performed to examine the Homeostasis Model Assessment-Insulin Resistance (HOMA-IR) Index among healthcare shift workers (HCSW). Materials and Methods: A total of 272 healthcare workers (HCWs) were invited to participate in the study within an occupational surveillance framework, 137 were HCSW while 135 were healthcare non-shift workers (HCNSW). Fasting glucose, insulin, and HOMA-IR Index were evaluated in each participant and correlated with shift workers. Results: Indicators of glucose metabolism were significantly higher in HCSW p < 0.001, and logistic regression analysis confirmed a significant positive association between increased values of HOMA-IR Index and shift workers (p < 0.05). Conclusions: Shift work could be a risk factor in developing insulin resistance and metabolic syndrome.

Serum concentrations of perfluorinated compounds among children living in Sicily (Italy).

Perfluorinated compounds (PFCs) are a large group of man-made chemicals characterized by a completely fluorinated hydrophobic linear carbon chain attached to a hydrophilic head. The presence of PFCs has been related with various adverse effects on several organs and systems. 61 children (6-11 years of age) living in Sicily underwent medical examination and blood and urine routinary tests, as well as the serum concentration of 16 PFCs. These compounds (∑PFCs = 5.888 ng/mL, range 1.035-12.94 ng/mL) were widely detected in the samples of all children, showing that they had been extensively exposed to PFCs, especially perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA), perfluorohexanesulfonate (PFHxs) and perfluorodecanoic acid (PFDA). In addition, long-chained PFCs were found in children's blood correlated with short-chained ones. Perfluorodecanoic acid (PFDA) (beta=-0.114) and PFHxS (beta = 0.003) levels were correlated with maternal parity; while the "months of breastfeeding" variable is correlated with perfluoroundecanoic acid (PFUnDA) concentrations (beta = 0.008). Female gender is negatively associated with PFOS (beta = 0.008). The adverse health effects of PFCs on young children should be noted, due to their growing phase. Therefore, further studies are needed to monitor and address adverse health outcomes of PFCs on children, especially those living in industrial regions.

Mutagenic and DNA repair activity in traffic policemen: a case-crossover study.

BACKGROUND: Emissions from vehicles are composed of heterogeneous mixtures of hazardous substances; several pollutants such as Polycyclic Aromatic Hydrocarbons (PAHs) are amongst the most dangerous substances detected in urban monitoring. A cohort of traffic policemen usually occupationally exposed to PAHs present in the urban environment were examined in order to assess the mutagenicity and DNA capacity repair. METHODS: Seventy-two urban traffic policemen working in Catania's metropolitan area were enrolled in the study. Two spot urine samples were collected from each subject during the whole working cycle as follows: sample 1 (S1), pre-shift on day 1; sample 2 (S2) post-shift on day 6. 1-hydroxypyrene (1-OHP) was measured to serve as an indirect exposure indicator. Urinary mutagenic activity was assessed through the plate incorporation pre-incubation technique with S9, using YG1024 Salmonella typhimurium strain over-sensitive to PAH metabolite. Concentrations of urinary 8-oxodG were measured using liquid chromatography tandem mass spectrometry. RESULTS: As regards the exposure to PAHs, results highlighted a statistically significant difference (p < 0.001) between pre-shift on day 1 and post-shift on day 6 levels. Mutagenic activity was detected in 38 (66%) workers on S1 and in 47 (81%) on S2. Also 8-oxodG analysis showed a statistically significant difference between S1 and S2 sampling. CONCLUSIONS: This study demonstrated that occupational exposure to pollutants from traffic emission, assessed via 1-OHP measurements in urine, may lead to DNA repair and mutagenic activity, in line with other studies.

MMP-9 as a Candidate Marker of Response to BRAF Inhibitors in Melanoma Patients With BRAFV600E Mutation Detected in Circulating-Free DNA.

The BRAFV600E mutation is associated with melanoma development and its detection in circulating-free DNA cannot be observed in all melanoma patients harboring this mutation in tumor specimens. Beside the circulating-free DNA BRAFV600E mutation, other markers of therapeutic response should be identified. Matrix metalloproteinase-9 (MMP-9) could be one of them as its role as indicator of invasiveness in melanoma have been explored. In this study, MMP-9 was evaluated in melanoma cells after treatment with dabrafenib. In vitro data were validated in 26 melanoma patients, of which 14 treated with BRAF inhibitor alone and 12 treated with both BRAF and MEK inhibitors, by ELISA assay and droplet digital PCR for measuring MMP-9 serum levels and circulating-free DNA BRAFV600E mutation, respectively. Statistical analyses were performed to evaluate the prognostic significance of MMP-9, progression-free survival (PFS) and overall survival (OS) according to the BRAFV600E mutation and MMP-9 levels. The performed analyses showed that MMP-9 and pEKR1-2 were statistically down-regulated in melanoma cells after treatment with dabrafenib. Circulating-free DNA BRAFV600E mutation was detected in 11 out of 26 melanoma patients showing higher levels of MMP-9 compared to those with undetectable BRAFV600E mutation. Furthermore, higher levels of MMP-9 and circulating-free DNA BRAFV600E mutation were associated with lower PFS and OS. Finally, the monitoring of therapy showed that MMP-9 significantly decreased at T1 and T2, but not at T-last, for the patients with detectable circulating-free DNA BRAFV600E mutation. In conclusion, high levels of MMP-9 and circulating-free DNA BRAFV600E mutation are associated with poor PFS and OS. MMP-9 may represent a promising indicator of response to BRAF inhibitors in combination with the detection of BRAFV600E mutation.

Occupational exposure to fluoro-edenite and prevalence of anti-nuclear autoantibodies.

An environmental contamination due to an asbestiform mineral fiber, fluoro-edenite (FE), caused a significantly increased mortality rate for malignant mesothelioma in Biancavilla, Italy. Exposure to fluoro-edenite has been associated with inflammatory processes as an early response to inhaled fibers. The aim was to explore prevalence of anti-nuclear autoantibodies (ANA) in a group of construction workers residing and working in the contaminated area. Prevalences for samples positive to ANA were 60% (n = 9) and 13% (n = 2), for exposed and nonexposed, respectively (p-value <0.05), the odds ratio was 9.75 (95% CI: 1.59-59.69). The significance of elevated ANAs in subjects exposed to fibers is unknown; additional studies may provide a better opportunity to establish a correlation between autoimmunity and environmental exposure.

Prevalence of anti-nuclear autoantibodies in subjects exposed to natural asbestiform fibers: a cross-sectional study.

Fluoro-edenite (FE) is an asbestiform mineral fiber spotted in the lava rocks excavated from a stone quarry in Biancavilla (Italy). The derived material had been employed locally for building purposes. Previous studies found evidence that exposure to asbestos may induce autoimmunity, with frequency of anti-nuclear autoantibodies (ANA). The aim of this study was to explore the relationship between FE exposure and autoimmune responses in an exposed population. For the study, 60 subjects living in the area of Biancavilla and 60 subjects as control group were randomly invited to participate. A free medical check, including spirometry and a high-resolution computer tomography chest scan, was given to all participants. ANA were determined by indirect immunofluorescence. On medical check, no subject showed any sign and/or symptoms of illness. Prevalence for samples positive to ANA were 70% (n = 42) and 25% (n = 15), respectively, for exposed and non-exposed subjects (p < 0.05). The presence of pleural plaques (PP) was found in 21 (30%) of the exposed subjects and in 2 (3%) of the non-exposed participants. PP subjects were always ANAs positive. In conclusion, as already it was observed with exposure to asbestos fibers, levels of ANA seemed to significantly increase in subjects who had been exposed to FE. Furthermore, all subjects showing PP were also ANA-positive. This first finding in subjects exposed to FE should encourage researchers to further investigate associations between autoimmune unbalance and environmental exposure to asbestiform fibers.

Silibinin Restores NAD⁺ Levels and Induces the SIRT1/AMPK Pathway in Non-Alcoholic Fatty Liver.

Nicotinamide adenine dinucleotide (NAD⁺) homeostasis is emerging as a key player in the pathogenesis of non-alcoholic fatty liver disease (NAFLD) and is tightly linked to the SIRT1/5'-AMP-activated protein kinase (AMPK) pathway. Silibinin, the main component of silymarin, has been proposed as a nutraceutical for the treatment of NAFLD. In this study, we aimed to identify whether silibinin may influence the NAD⁺/SIRT1 axis. To this end, C57BL/6 mice were fed a high fat diet (HFD) for 16 weeks, and were treated with silibinin or vehicle during the last 8 weeks. HepG2 cells were treated with 0.25 mM palmitate for 24 h with silibinin 25 µM or vehicle. HFD and palmitate administration led to oxidative stress, poly-(ADP-ribose)-polymerase (PARP) activation, NAD⁺ consumption, and lower SIRT1 activity. In mice fed the HFD, and in HepG2 treated with palmitate, we consistently observed lower levels of phospho-AMPKThr172 and phospho-acetyl-CoA carboxylaseSer79 and higher levels of nuclear sterol regulatory element-binding protein 1 activity, indicating de novo lipogenesis. Treatment of mice and HepG2 with silibinin abolished oxidative stress, and inhibited PARP activation thus restoring the NAD⁺ pool. In agreement with preserved NAD⁺ levels, SIRT1 activity and AMPK phosphorylation returned to control levels in mice and HepG2. Our results further indicate silibinin as a promising molecule for the treatment of NAFLD.

The risk of HCV infection among health-care workers and its association with extrahepatic manifestations (Review).

Health care workers (HCWs) are frequently exposed to different biological agents during their activities and are frequently monitored. Among these infectious agents, human hepatitis C (HCV) can infect HCWs. In this review article, the risk of HCV infection among HCWs is discussed along with extrahepatic HCV-related malignancies, such as non­Hodgkin lymphoma. Accidental contamination, represented by percutaneous and mucocutaneous infections is the main risk factor for such infection. The compliance of the protection procedures, included in the current regulation for HCWs, is the most important issue to reduce the risk of pathogen infections that in turn may produce reduction of infection­associated malignancies.

BRAF mutations in papillary thyroid carcinoma and emerging targeted therapies (review).

Papillary thyroid carcinoma (PTC) is the most common histotype among the thyroid cancer types. Although PTC is a curable malignancy, many patients relapse after treatment. Thus, there is a need to identify novel factors involved in the pathogenesis of PTC that may be used as targets for new therapies. The MAPK pathway has been implicated in the pathogenesis of PTC. Therefore, in this review, we summarize the role of the BRAF V600E mutation in the development and progression of thyroid cancer. The cinical implication of this molecular abnormality is also discussed. It is evident that the detection of the BRAF V600E mutation is crucial in order to identify novel avenues for thyroid cancer treatment.

microRNAs and thyroid cancer: biological and clinical significance (Review).

Despite recent advances in the management of thyroid cancer, the survival rate of this tumor may still be improved. Therefore, the identification of biological and molecular features of indolent and aggressive disease would be critical to define clinically useful predictors of high-risk lesions. microRNAs (miRNAs) are small RNA molecules with regulatory function and marked tissue specificity that modulate multiple targets belonging to several pathways. They are frequently deregulated in cancer and constitute a new class of blood-based biomarkers useful for cancer detection and prognosis definition, including thyroid cancer. In this review, the role of miRNAs in thyroid cancer development is described. The most common miRNAs detected in thyroid cancer along with their clinical significance are also discussed. Further studies aimed to detect plasma-based miRNA biomarkers in thyroid cancer patients may provide further insight into the management of thyroid cancer.

Extrahepatic disorders of HCV infection: a distinct entity of B-cell neoplasia?

Some infectious agents have been associated with B-cell lymphoma development. In the last decades, it has been demonstrated that patients infected by hepatitis C virus (HCV) are more likely to develop B-cell non-Hodgkin's lymphoma (NHL) than those uninfected. The prevalence of HCV-infection among NHL patients is reported in this review of all Italian studies on NHL and HCV infection, both case-control and case series. From 18 studies, the prevalence of anti-HCV antibodies in 2736 NHL patients was 19.7% (range: 8.3-37.1%). The association of HCV-infection with each NHL histotype in case-control studies is discussed. Molecular mechanisms by which HCV infection promotes B-cell NHL development is also explored and indicate that HCV-associated lymphomas may be a distinct entity. Clarification of these mechanisms may improve diagnosis, classification and therapy of this subset of NHL. Finally, treatment of HCV-positive patients with lymphoproliferative disorders are herein summarized and further support the notion that HCV infection contributes to the development of these pathologic conditions.