Epidemiology of the new HIV infections in a large university hospital of Southern Italy: the importance of increasing the culture of the HIV screening test in the prevention of late diagnosis.

BACKGROUND: the delay of HIV diagnosis represents an important risk of spreading HIV infection within the community and, at the same time, a loss of opportunity for undiagnosed subjects to start the antiretroviral therapy. OBJECTIVES: to evaluate the difference, over time, between early and late HIV diagnosis in a large University Hospital of Southern Italy and to emphasize the importance of spreading the culture of prevention, based on the improvement of the HIV screening test adherence, in order to reduce the incidence of late HIV diagnosis. DESIGN: retrospective cross-sectional study. SETTING AND PARTICIPANTS: all the HIV screening tests performed in the six-year period 2013-2018 by the HIV laboratory of a third-level University hospital of Sicily (Southern Italy) were considered. The tests were performed on four categories of patients: voluntary HIV screening participants, inpatients, outpatients, and healthcare workers. MAIN OUTCOME MEASURES: number of performed HIV tests and frequency of early and late HIV diagnosis in the studied categories of subjects. RESULTS: in the considered period, 16,290 HIV tests were performed and the new diagnosis, considering all the four categories of patients, were in total 72, of which the highest percentage (45.8%) concerned voluntary HIV screening participants showing a mean CD4+ level >350/µL (threshold to discriminate early or late infection), followed by inpatients (27.8%) and outpatients (26.4%) with a mean CD4+ levels <350/µL. Moreover, from 2013 to 2018, the detection of serological positivity on voluntary HIV screening participants showed a decrease of 12.5%, while there was a parallel increase of 18.2% in the inpatients group. In the outpatients, the serological positivity remains quite stable. Concerning sexual habits, in the voluntary HIV screening participants, more than half (55.5%) of the HIV positive subjects were homo-bisexuals, while in the inpatients and outpatients' groups the highest percentage (83.3%) were heterosexuals. CONCLUSIONS: in this study, the majority (45.8%) of the new HIV diagnosis were detected on voluntary HIV screening participants in an earlier phase of infection. However, adding the percentages of inpatients and outpatients, it results that more than half (54.2%) of the new diagnosis occurred in a more advanced phase of infection. For these reasons, it appears necessary to stress the importance of an early diagnosis, reachable only by the spread of an HIV screening culture through health education campaigns addressed to the entire population and, especially, to heterosexual category that was the most interested group in the late diagnosis.

Ex vivo evaluation of the effects of a white grape juice extract on lymphocytic mitochondrial functions.

The physio-pathological role of mitochondria in aging and age-related diseases has stimulated the search for compounds able to promote mitochondrial functionality. Our study was designed to evaluate the effect of a white grape juice extract (WGJe) on mitochondrial activity, in an ex vivo experimental model consisting of activated lymphocytes obtained from a younger age group and an older age group of subjects. WGJe steadily decreased the lymphocytic mitochondrial mass in the older subjects, without a relevant effect in their younger counterpart, and significantly enhanced Δψm in both groups investigated. Finally, WGJe reduced the endogenous mitochondrial production of H2O2 in all subjects. The results support the potential use of WGJe to improve mitochondrial functionality, thus maintaining human health and slowing down aging.

The contribution of professors of Hygiene to the progress and development of public health in Italy: one hundred years of history.

In 1917 Achille Sclavo, a distinguished researcher and founder of the Italian Society of Hygiene, ended up the first term as Rector of the University of Siena. Since then, the contribution of professors of hygiene in Italian universities has ranged over several relevant topics including vaccinations, environmental hygiene, hospital hygiene, healthcare organization and management, with an important contribution to the health reform of 1978 by Augusto Giovanardi and Alessandro Seppilli. Several Academic Schools (Roman, Neapolitan, Genoese, Sicilian, Venetian, Lombard, etc.) have produced excellent researchers, teachers and mentors who have also occupied important positions in the panorama of the Italian health system. This note analyzes the main research topics, the most famous institutes and departments of hygiene and public health and the contributions of the most famous professors for the development of the discipline, the management of the post- graduated Schools in hygiene and preventive medicine, the commitment in scientific associations and the role of some of them in important institutional positions. Even through its contribution and constant commitment to the institutions, Italian public health has achieved the reputation of being one of the best known and appreciated in the international scientific community.

Vaccine hesitancy: An overview on parents' opinions about vaccination and possible reasons of vaccine refusal.

Background. Vaccine hesitancy has increased worldwide with a subsequent decreasing of vaccination rates and outbreaks of vaccine-preventable diseases (i.e. measles, poliomyelitis and pertussis) in several developed countries, including Italy. Design and Methods. We conducted a survey to investigate the attitudes of a parents' sample about vaccinations by the distribution of questionnaires in six lower secondary schools of the Italian city of Messina. Results. Regarding vaccinations carried out on children, the declared vaccination coverage rates ranged widely between good coverage percentages for some vaccinations (Measles-Mumps-Rubella, Diphtheria-Tetanus-Pertussis), and very low coverage rates for others, especially for "new" vaccinations (HPV, meningococcal, pneumococcal). The vaccinations carried out correlated negatively with both parents' age and their level of education. Moreover, a favourable parents' opinion was strongly influenced by a favourable opinion of the physician, while an unfavourable parents' opinion seemed conditioned by a direct or indirect knowledge of people harmed by vaccines. In addition, our data show that parents do not often know or partially know the real composition of the vaccines and the diseases prevented by vaccinations. Conclusions. Data analysis shows that parents are, theoretically, favourable towards vaccinations but have little knowledge of such practices, sometimes not being unaware of the types of vaccines administrated to their children. Health education and communication of correct information are certainly the cornerstones to improve the situation and to fight the widespread and non-grounded fears about vaccines.

Brucellosis is a public health problem in southern Italy: Burden and epidemiological trend of human and animal disease.

BACKGROUND: Brucellosis is the most common global bacterial zoonotic disease. Italian annual 2015 report on animal brucellosis control that was submitted to the Integrated National Plan of the Italian Health Ministry showed that the Italian region with the highest prevalence and incidence of brucellosis was Sicily (3.3%). This study aims to demonstrate the burden of disease and the epidemiological trend of human and animal brucellosis in Messina, Sicily, from 1997 to 2016. METHODS: The analysis was conducted in the twenty-years 1997-2016. We examined the computerised and paper registers of the Messina Provincial Health Agency n.5 to evaluate human and animal brucellosis reports. RESULTS: 1462 cases of human brucellosis were reported with an important outbreak in 2016 in which were reported 137 cases while the prevalence of infected cattle and sheep/goats decreased from 3.8% and 8%, respectively, in 1997 to 1.7% for both in 2016. A statistically significant correlation was observed between the decrease of both animal and human cases during the considered period. CONCLUSIONS: Our study demonstrate that brucellosis is still present in Sicily with a number of cases identified in both animals and humans and it hypothesises a large number of probable underreported cases. Our findings confirm the need to improve knowledge of the risks associated with consuming raw milk and its derivatives, mainly from sheep and goats, and demonstrates that public health would benefit from cooperation between human and veterinary health services.

Head and neck squamous cell carcinoma and its correlation with human papillomavirus in people living with HIV: a systematic review.

Over the last 20 years we assisted to an increase in the mean age of People Living with HIV and their comorbidities. Especially, there was an increase in Human Papillomavirus-related head and neck squamous cell carcinomas. Despite their increasing incidence in HIV-positive people, mechanisms that lead to their development and progression are only partially understood. The aim of this review is to identify key data and factors about HPV-related head and neck squamous cell carcinoma in HIV-seropositive patients. Systematic search and review of the relevant literature-peer-reviewed and grey-was conducted using the Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA) guidelines. We included in our review only the 35 full-text articles we considered the most substantial. It is mandatory to improve our knowledge about the interactions existing between HPV and HIV, and about their actions on oral mucosa immune system.

In vitro assessment of the indirect antioxidant activity of Sulforaphane in redox imbalance vanadium-induced.

Owing to sulforaphane presence, a dietary consumption of Brassicaceae prevents chronic diseases. This hormetic compound induces adaptive stress response at subtoxic doses, while doses that exceed the cellular defence are toxic. In HepG2, Caco-2 and Vero cells, we investigated the sulforaphane (SFN) (5 µM) role in counteracting redox imbalance induced by VOSO4 [V(IV)]. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test showed a dose-dependent viability reduction (r < -0.95; p < 0.01) (range 5-80 µM). At 5 µM, SFN enhancement of mitochondrial activity was confirmed by Δψm (p < 0.05) both in basal condition and in redox-stressed cells. Intracellular ROS, DNA and lysosomal oxidative damages underlined the indirect antioxidant SFN activity, confirmed by the increase of GSH. The SFN empowering effects on mitochondrial function were imputable to the presence of mitochondrial proteins among the Nrf2-responsive phase II proteins. Considering the link between oxidative stress and chronic diseases, a long-term dietary intake of Brassicaceae could be strongly advisable.

Prevalence of human papillomavirus in saliva of women with HPV genital lesions.

BACKGROUND: The human papilloma viruses (HPVs) are DNA viruses associated with benign and malignant lesions of skin and mucous membranes. The HPVs has been implicated as the cause of virtually all cervical cancers worldwide but studies showed that these viruses can cause numerous cancers in several tissues including Oral Squamous Cell Carcinoma (OSCC). At least 90 % of HPV-positive OSCCs are associated with high-risk (or oncogenic) HPV-16 and oral infection confers an approximate 50-fold increase in risk for HPV-positive OSCC. HPV-positive OSCCs are associated with sexual behaviors in contrast to HPV-negative OSCCs that are associated with chronic tobacco and alcohol use. The aim of this study was to estimate the prevalence of HPV-DNA in saliva samples collected from women in which it has been previously established the HPV infection of the cervix with relative genotyping and, then, to study the possible correlation. METHODS: Saliva samples were collected from 100 women with HPV cervical lesions, aged between 22 and 52 years old, and 25 healthy women with normal cytology (control group), aged between 20 and 49 years old. PCR assay was used to detect HPV DNA. RESULTS: The prevalence of oral HPV infection in saliva samples was 24 % in women with HPV cervical lesions while in the control group was 8 %. It has been demonstrated a strong association between high grade squamous intraepithelial lesion and oral infection due to HPV16 and 18, that are the most frequently detected HPV genotypes. CONCLUSION: This study shows that patients with genital HPV infection are at risk for oral infection and, consequently, for the development of OSCC.

Higher levels of oxidative DNA damage in cervical cells are correlated with the grade of dysplasia and HPV infection.

The Human papillomavirus is responsible for the most common sexually transmitted infection and is also known to be an oncogenic virus that is associated with cervical, anogenital, and head-neck cancers. The present study aims to assess whether oxidative DNA damage is correlated with the grade of HPV-related lesions. Moreover, we evaluated clinical data and unhealthy lifestyles to verify their possible influence on the genesis of oxidative DNA damage in cervical cells. We quantified the amount of 8-Oxo-2'-deoxyguanosine in DNA as a biomarker of oxidative damage in women with and without HPV infection. We also correlated oxidative damage with different stages of cervical lesions and available clinical data (e.g., HPV genotypes). To identify HPV infections, in which proteins with a transforming potential are produced, we performed a qualitative detection of HPV E6/E7 mRNA. Our results showed greater oxidative damage in HPV-related dysplastic cervical lesions compared to samples with normal cytology, especially in women with high-grade squamous intraepithelial lesions. The latter showed a closed link with high-risk HPV genotypes. Reactive oxygen species can induce DNA double-strand breaks in both the host DNA and in the circular viral episome; this could facilitate the integration of the virus, promoting HPV carcinogenesis. Therefore, in HPV-infected women, it could be useful to reduce additional resources of reactive oxygen/nitrogen species (RONS) with a healthy lifestyle.

Mitochondrial-mediated apoptosis pathway in alveolar epithelial cells exposed to the metals in combustion-generated particulate matter.

Previously a significant mitochondrial impairment was identified in alveolar epithelial cells exposed to metals adsorbed to combustion-generated particulate matter (PM). Due to the critical role of mitochondria in apoptosis, the aim of this study was to investigate the pro-apoptotic potential of metals present in oil fly ash (OFA). A549 cells were exposed to water-soluble components of an OFA sample, containing vanadium [V(IV)], iron [Fe(III)], and nickel [Ni(II)] (68.8, 110.4, and 18 µM, respectively). Experiments were also performed using individual metal solutions. Apoptosis was detected and the mitochondrial role was assessed by a caspase-9 inhibitor (Z-LEHD-FMK). To determine whether the presence of impaired mitochondria in unexposed daughter cells increased apoptosis, an in vitro model was developed that allowed determination of effects until the third cell generation. To specifically examine the toxicity of vanadium (V), that characterize the airborne pollutant examined in this study, p53involvement and metabolic impairment through changes in HIF-1α and Glut-1 expression were determined. OFA and individual metal solutions produced significant apoptosis in the progeny of exposed cells, triggering the intrinsic apoptosis pathway. In apoptosis induced by poorly genotoxic metal V, p53 did not play a significant role. However, V exposure increased nuclear translocation of HIF-1α and expression of the Glut-1 receptor, indicating metabolic impairment due to metal-induced mitochondrial dysfunction. Overall, these results improve our knowledge of the pathogenic role that airborne metals and in particular V exerted in respiratory epithelium.

Mitochondrial dysfunction by pro-oxidant vanadium: ex vivo assessment of individual susceptibility.

The aim was to assess the individual susceptibility to mitochondrial impairment induced by ex vivo exposure to vanadium, an airborne pro-oxidant pollutant. In lymphocyte cultures V(IV)-treated of forty-five healthy subjects, we evaluated the mitochondrial transmembrane potential (Δψm) and the H2O2 in comparison to background values. As variables, we included both lifestyle factors and genetic polymorphisms (GSTM1 and GSTT1 variants, and C677T and A1298C variants of methylenetetrahydrofolate reductase MTHFR). H2O2 mitochondrial content increased significantly (P<0.05) after metal exposure while, in comparison to basal Δψm, both depolarisation and hyperpolarisation were recorded. This underlined the mitochondrial dysfunction vanadium-induced that worsens the redox imbalance by endogenous ROS overproduction. Only age was found to contribute significantly to the high inter-individual variability, as assessed by multivariate analysis. In older subjects, the H2O2/Δψm values underline the organelle impairment and, under V-exposure, Δψm values were inversely related to age (R=-0.591; P=0.012).

HIV genomic mutations causing resistance to antiretroviral drugs in seropositive Sicilians.

Through the use of highly active antiretroviral therapy a significant reduction occurred in mortality and morbidity caused by Human Immunodeficiency Virus. The use of antiretroviral drugs resulted in the emergence of resistant viral strains due to mutations that cause a selective advantage to the virus. The aim of our study is to monitor the HIV-1 infection in Sicilians patients evaluating the presence of mutations that make the virus resistant to the therapy. The QIAGEN QIAamp Viral RNA Mini Kit was used to extract HIV-1 viral RNA from 300 patients while the TRUGENE HIV-1 Genotyping Kit and the OpenGene DNA Sequencing System determined viral mutations in the RNA samples. The analysis showed that from 300 subjects, 116 developed Antiretroviral Drug Resistance. The percentage of patients with resistance to nucleoside reverse transcriptase inhibitor (NRTI), non nucleoside reverse transcriptase inhibitor (NNRTI) and protease inhibitor was 26%, 23% and 20%, respectively. Comparison between drug resistances and mutations showed that 134 individuals had mutations in genes codifying for reverse transcriptase but a little more than 50% were associated with resistance to reverse transcriptase inhibitors, in particular 78 and 68 subjects developed drug resistances to NRTI and NNRTI classes respectively. Subjects that showed mutations in genes codifying for protease were 216 but only 59 of these were associated with resistance to protease inhibitors. Our findings emphasize the importance of continued resistance surveillance. Monitoring of transmitted resistance continues to be needed among treatment-exposed patients because of the benefit it provides for the development of drugs effective against the most frequently found drug-resistant viruses.

Bioenergetics of T cell activation and death in HIV type 1 infection.

Regressive morphological lesions, found in peripheral lymphocytes from HIV(+) patients, clearly conflict with normal cycle progression and with the execution of basic housekeeping and immune functions. With these lesions, circulating lymphocytes are destined to spontaneous and energy-independent cell lysis. By means of confocal microscopy and morphometry, we have quantified the rate of circulating T cells that are probably destined to emocatheresis in vivo. This rate includes lymphocytes in which nucleolin fragments have been scattered out of the nuclear region as a result of prelethal alterations in the nuclear membrane permeability. In terms of bioenergetics, these cells show evident anomalies in the energy production machinery that make them unable to carry out ATP-requiring functions. The extent of damaged cell fraction in peripheral blood reflects the frequency with which T lymphocytes leave lymphoid tissue to be cleared in hemocatheretic processes.

Intracellular accumulation of cell cycle regulatory proteins and nucleolin re-localization are associated with pre-lethal ultrastructural lesions in circulating T lymphocytes: the HIV-induced cell cycle dysregulation revisited.

The HIV-induced demise of CD4-T cells is thought to be a result of the execution of genetically programmed cell death that occurs in lymphoid tissue, where many resident T cells are chronically hyperactivated. Since HIV-induced alterations of cell cycle control has been often indicated as prominent mechanism of immune hyper activation and cause of apoptotic death, the signal pathway involved in cell cycle dysregulation of T lymphocytes from HIV infected patients was extensively studied. Here, we also demonstrate that circulating T lymphocytes leave lymphoid tissues with diffused regressive lesions (vacuolization, blebbing, nuclear evanescence and organelle swelling). Equally diffused are biochemical anomalies that accompany the overall disarrangement of cell structure, particularly the fragmentation and diffusion into the cytoplasm of C23/nucleolin, the intracellular accumulation of short lived regulatory proteins and the decrease in expression of membrane proteins. All this is something more than a cell cycle-related remodelling of cell morphology and biochemical mechanisms, and rather recalls a necrotic/oncotic cell damage. Since these changes are associated with adaptive mechanisms to hypoxia, we give evidence for alteration of cell cycle control developing in conditions of scarce energy supply.

Homocysteine induces DNA damage and alterations in proliferative capacity of T-lymphocytes: a model for immunosenescence?

Homocysteine (Hcy) appears to exert different effects on immune functions possibly contributing to age-related pathological states, including vascular diseases, immune dysfunction, and Alzheimer's disease. However, molecular mechanisms underlying Hcy toxicity need to be better characterized. Since T cells are a suitable model to address the possible role of replicative senescence during the in vivo aging, we investigated the effects of high Hcy concentrations on mitogen-activated lymphocytes, with regard to evaluation of DNA damage and cell cycle alterations. Cultured human peripheral blood lymphocytes were stimulated with mitogenic concanavalin A (5 microg/ml) for 48 h in the presence or absence of Hcy (1 mM). Both flow cytometric analysis and caspase-3 activity assay showed an increased rate of apoptosis in Hcy-treated lymphocyte cultures compared to controls. Further, Hcy exposure caused DNA fragmentation as evaluated by single cell gel electrophoresis showing the occurrence of comets. Cytokinesis-block micronucleus assay, performed after addition of cytochalasin B (5 microg/ml) and incubation up to 72 h, revealed a significantly higher frequency of micronucleated/binucleated cells in Hcy-treated cultures compared to controls (P < 0.001). Hcy also reduced cyclin B expression in comparison to control cultures, while cyclin D levels were not significantly affected. Cell cycle alterations, such as the inability of cells to enter into mitosis, could be related with DNA damage. These findings provided a link between perturbation of lymphocyte proliferation homeostasis and commitment towards apoptosis. Our results suggest the involvement of Hcy in the altered immune function associated with age and disease pathology.

Prevalence of SENV-H and SENV-D virus: epidemiological study in blood donors and dialysis patients.

INTRODUCTION: Recently, the identification of the SEN virus as a possible etiological agent of parental transmission hepatitis led to the study of the prevalence of such pathogen agents, particularly SENV-H, in our population. This paper compares the rate prevalence in high-risk subjects, such as dialysis patients, and low-risk subjects, such as blood donors. MATERIAL AND METHODS: The study was carried out on SEN virus DNA extracted from serum of dialysis patients and blood donors, and the presence of viral genomes was performed by the nested PCR method. RESULTS: The results showed a higher prevalence in male blood donors, supporting the hypothesis of an epidemiological role for sexual and also parental transmission, as is clearly demonstrated by the high prevalence in dialysis patients. The result reduced the importance of the possible etiological role of the SEN virus due to the high percentage of positivity in healthy population, and it induces one to consider poorly significant the pathogenicity of such viral agents. CONCLUSION: For this instance, the authors, in agreement with the phylogenically related TT virus, described SEN viruses as absolutely not pathogens and considered them as "simple guests."

Aeromonas hydrophila exotoxin induces cytoplasmic vacuolation and cell death in VERO cells.

Many organisms are able to cause cell vacuolation, but it is unclear if this can be considered a step of apoptosis or necrosis, or a distinct form of cell death. In this study VERO cells were used to evaluate the relationship between vacuolation and cell death pattern caused by exotoxins produced by environmental strains of A. hydrophila. Cell damage has been evaluated morphologically as well as biochemically. Cytotoxic and vacuolating titres were strictly correlated and the vacuolation has to be considered an early indicator of cytotoxicity that causes cell apoptosis or necrosis in relation to the dose. Signs of apoptosis (chromatin condensation and blebbing) were observed at low concentration and TGase activity, referable to apoptosis induction, confirms morphological observations. In fact, putrescine incorporation was related both to cytotoxin concentration and time of incubation. Moreover, the observed doubling cells with necrotic features permit us to suppose that cell sensitivity and death pattern could change during the different phases of cellular cycle.