Importance of oxygen tension in human ovarian tissue in vitro culture.

STUDY QUESTION: Is there any difference between 20% and 5% oxygen (O2) tension in vitro culture (IVC) on the viability and quality of human follicles contained in cultured ovarian cortex? SUMMARY ANSWER: An O2 tension of 5% yields higher follicle viability and quality than does 20% O2 tension after 6 days of IVC. WHAT IS KNOWN ALREADY: The primordial follicle (PMF) pool resides within the ovarian cortex, where the in vivo O2 tension ranges between 2% and 8%. Some studies suggest that lowering O2 tension to physiological levels may improve in vitro follicle quality rates. STUDY DESIGN, SIZE, DURATION: This prospective experimental study included frozen-thawed ovarian cortex from six adult patients (mean age: 28.5 years; age range: 26-31 years) who were undergoing laparoscopic surgery for non-ovarian diseases. Ovarian cortical fragments were cultured for 6 days at (i) 20% O2 with 5% CO2 and (ii) 5% O2 with 5% CO2. Non-cultured fragments served as controls. PARTICIPANTS/MATERIALS, SETTING, METHODS: Cortical fragments were used for the following analyses: hematoxylin and eosin staining for follicle count and classification; Ki67 staining to evaluate PMF proliferation; cleaved caspase-3 immunostaining to identify follicle apoptosis; 8-hydroxy-2-deoxyguanosine and gamma-H2AX (γH2AX) immunolabeling to detect oxidative stress damage and DNA double-strand breaks (DSBs) in oocytes and granulosa cells (GCs); and ß-galactosidase staining to assess follicle senescence. Droplet digital PCR was also performed to further explore the gene expression of superoxide dismutase 2 (SOD2) and glutathione peroxidase 4 (GPX4) from the antioxidant defense system and cyclin-dependent kinase inhibitors (p21 and p16) as tissue senescence-related genes. MAIN RESULTS AND THE ROLE OF CHANCE: Apoptosis (P = 0.002) and follicle senescence (P < 0.001) rates were significantly lower in the 5% O2 group than in the 20% O2 group. Moreover, GCs in follicles in the 20% O2 group exhibited significantly (P < 0.001) higher oxidative stress damage rates than those in the 5% O2 group. DNA DSB damage rates in GCs of follicles were also significantly higher (P = 0.001) in the 20% O2 group than in the 5% O2 group. SOD2 expression was significantly greater in the 5% O2 group compared to the 20% O2 group (P = 0.04) and the non-cultured group (P = 0.002). Expression of p21 was significantly increased in both the 20% O2 (P = 0.03) and 5% O2 (P = 0.008) groups compared to the non-cultured group. Moreover, the 20% O2 group showed significantly greater p16 expression (P = 0.04) than the non-cultured group, while no significant variation was observed between the 5% O2 and no culture groups. LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: This study focuses on improving follicle outcomes during the first step of ovarian tissue IVC, where follicles remain in situ within the tissue. The impact of O2 tension in further steps, such as secondary follicle isolation and maturation, was not investigated here. WIDER IMPLICATIONS OF THE FINDINGS: Our findings suggest that 5% O2 tension culture is a promising step toward potentially solving the problem of poor follicle viability after IVC. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by grants from the Fonds National de la Recherche Scientifique de Belgique (FNRS-PDR T.0064.22, CDR J.0063.20 and grant 5/4/150/5 awarded to M.M.D.). The authors have nothing to disclose.

A cross-sectional study on smartphone uses among pregnant women attending childbirth classes in the Metropolitan Area of Palermo, Italy: The Stop-Phone study.

Background: Prevalence of mobile device addiction has increased over the years; both women and men have assimilated the mobile phone as a central component of their personal existence: integrating it into their lifestyle or becoming so dependent on it that life without it has become unimaginable. Smartphones generate radio-frequency electromagnetic fields. While short-term exposure in adults was considered quite safe, effects of long-term exposure or exposure during pregnancy on fetuses or during breastfeeding on newborns are not well studied yet. The objective of the present study was to investigate the prevalence and usage characteristics of smartphones among a sample of pregnant women, and promote the correct and conscious use of the smartphone. Methods: A cross-sectional study was conducted, with a questionnaire administered during childbirth classes and - after the questionnaire administration - an educational intervention focused on promoting the correct and conscious use of smartphones was carried out by psychologists and psychotherapists. Results: The findings of our study suggest that a significant number of the participants suffered addiction to mobile phone usage, but were not aware of it. More than two third of the sample (67.2%) have not changed their smartphone use habits since the beginning of their pregnancy and even more significant data shows that almost all future moms (98.3%) never speak with their doctor about smartphone use during pregnancy. Conclusions: Data collected suggest a lack of attention to the proposed topic, especially in relation to pregnancy. It seems necessary to sensitize future mothers on this topic. The promotion of a more conscious and controlled use of electronic devices can help reduce the radiation to which the unborn child may be exposed, but has a fundamental role even after birth, to ensure an adequate psychomotor and relational development of the child and do not affect, due to uncontrolled use of smartphones, the mother-child relationship.

Knowledge, attitudes and adherence towards influenza and other vaccinations among healthcare workers at the University Hospital of Palermo, Italy, during the first COVID-19 pandemic season (2020/2021).

Background: Increasing adherence to influenza vaccination among healthcare workers is a public health priority, stated that actually remains far below than international recommendations. During the 2020/2021 pandemic season, COVID-19 vaccines were not yet available until the end of December 2020, and influenza vaccines were the only one available to protect against seasonal respiratory diseases. The main objective of the present study was to assess knowledge, attitudes and adherence to influenza and other vaccinations recommended by the National Immunization Plan 2017-2021 for healthcare workers. Methods: Enrollment lasted from October and December 2020 at the vaccination unit of the University Hospital of Palermo. Data were collected through an anonymous and self-administered questionnaire, divided into 5 sections and 31 items. Results: Among 734 healthcare professionals that completed the survey, a significantly higher adherence to influenza vaccination was observed among healthcare workers that were more prone to receive COVID-19 vaccination (OR=4.02; 95% CI: 1.63-9.91). Moreover, higher influenza vaccination rates were observed among healthcare professionals that received influenza vaccination during previous 2019/2020 season (OR=15.3; 95% CI: 5.17-45.1) and that were favorable to the possible impact on increasing adherence of influenza mandatory vaccination (OR=4.88; 95% CI: 2.43-9.80). Conclusions: Propensity of healthcare workers to undergo vaccinations recommended in the National Immunization Plan increased during the first pandemic season. At the end of the vaccination season, flu vaccination coverage reached highest rates ever at the University Hospital of Palermo (around 60%), remaining anyway below the recommended minimum value of 75%. During next seasonal flu vaccination campaigns, it becomes essential to promote communication and information strategies to increase flu vaccination among healthcare workers, also focusing on co-administration with the anti-COVID-19 booster/seasonal doses.

The Erice 58 Charter on 'The health of migrants. An equity challenge for the public health system'.

Abstract: The Erice 58 Charter titled "The Health of Migrants: a Challenge of Equity for the Public Health System", was unanimously approved at the conclusion of the 58th Residential Course of the School of Epidemiology and Preventive Medicine 'Giuseppe D'Alessandro' entitled "The Health of Migrants: a Challenge of Equity for the Public Health System. Epidemiological, clinical-relational, regulatory, organisational, training and public communication aspects at international, national and local level', which took place from 28 March to 2 April 2022 in Erice (Sicily, Italy), at the 'Ettore Majorana' Foundation and Centre for Scientific Culture. The Course was promoted by the Italian Society of Migration Medicine (S.I.M.M.) and the Italian Society of Hygiene, Preventive Medicine and Public Health (SItI), with the collaboration and patronage of the Istituto Superiore di Sanità (ISS). 72 learners participated (mainly resident doctors in 'Hygiene and Preventive Medicine' but also other health service professionals), whose average age was 37 years; on the basis of territorial origin, 13 of the 20 Italian regions were represented. During the intense learning experience, which consisted of 18 frontal lessons (with 20 lecturers from the bio-medical, socio-anthropological and journalistic fields) and 7 working group sessions (supported by 4 classroom tutors in addition to the lecturers) in 'blended learning' mode, the various dimensions and critical issues related to the possibility of guaranteeing truly inclusive health policies for foreigners/migrants, throughout the country, were identified and discussed from an 'Health Equity' perspective. This enabled a small editorial group to draw up the basic document that, in the last session of the Course, was discussed and modified by all participants into the version of the 'Erice 58 Charter' presented here.

Impact of the environment on the health: From theory to practice.

The Erice 56 Charter titled "Impact of the environment on the health: from theory to practice" was unanimously approved at the end of the 56th course of the "International School of Epidemiology and Preventive Medicine G. D'Alessandro" held from 3rd to November 7, 2019 in Erice - Sicily (Italy) and promoted by the Study Group of "Environment and Health" of the Italian Society of Hygiene, Preventive Medicine and Public Health. The course, that included lectures, open discussions and guided working groups, was aimed to provide a general training on epidemiological and toxicological aspects of the environmental health impact, to be used by public health professionals for risk assessment, without forgetting the risk communications. At the end of the course 12 key points were agreed among teachers and students: they underlined the need of specific training and research, in the perspective of "One Health" and "Global Health", also facing emerging scientific and methodological issues and focusing on communication towards stakeholders. This Discussion highlight the need to improve knowledge of Health and Environment topic in all sectors of health and environmental prevention and management.

Use of a comprehensive diagnostic algorithm for Anisakis allergy in a high seroprevalence Mediterranean setting.

Summary: Background.Diagnosis of anisakis allergy (AA) is based on the skin prick test (SPT) and specific IgE (sIgE) determination. Anyway, false positivity cases are due to cross reactivity with numerous allergens. The aim of the study was to evaluate the reliability of a comprehensive diagnostic algorithm for the AA. Methods.An observational study was conducted on a sample of consecutive subjects accessing the allergology outpatient ambulatories of two hospitals located in Western Sicily. All the recruited outpatients were tested by Skin Prick Test performed using anisakis extracts by ALK-Abellò (Madrid, Spain). Specific IgE dosage for anisakis extracts was then performed by using ImmunoCAP250 (Immunodiagnostics Uppsala, Sweden). Consequently, outpatients who tested positive to first line tests underwent sIgE testing for ascaris and tropomyosin. Lastly, outpatients positive to the first line were invited to be further tested by basophil activation test (BAT) by using Flow CAST kit and anisakis commercial extract (Bühlmann Laboratories AG, Schönenbuch, Switzerland), as confirmatory analysis. Results.One hundred and eleven outpatients with an anamnesis suggestive of sensitization to anisakis (AS) and 466 subjects with chronic urticaria (CU) were recruited in the study. Of these, 22 with AS and 41 with CU showed a sensitization to anisakis allergens. The diagnostic algorithm revealed that 8.8% of outpatients who tested positive to sIgE determination were affected by CU, while 82.5% of all the sIgE positivity was related to cross-reactivity. Overall, a genuine anisakis seroprevalence of 2.3% was documented. Within a sub-sample of 15 subjects with clinical symptoms related to AA, n. 8 showed a real positivity after BAT. A greater response to A. pegreffii allergens as compared to A. simplex was reported. Conclusions.Our preliminary findings support the high clinical specificity of BAT for AA diagnosis, suggesting implementing this method in a comprehensive diagnostic algorithm.

Neurophysiology of the pedunculopontine tegmental nucleus.

The interest in the pedunculopontine tegmental nucleus (PPTg), a structure located in the brainstem at the level of the pontomesencephalic junction, has greatly increased in recent years because it is involved in the regulation of physiological functions that fail in Parkinson's disease and because it is a promising target for deep brain stimulation in movement disorders. The PPTg is highly interconnected with the main basal ganglia nuclei and relays basal ganglia activity to thalamic and brainstem nuclei and to spinal effectors. In this review, we address the functional role of the main PPTg outputs directed to the basal ganglia, thalamus, cerebellum and spinal cord. Together, the data that we discuss show that the PPTg may influence thalamocortical activity and spinal motoneuron excitability through its ascending and descending output fibers, respectively. Cerebellar nuclei may also relay signals from the PPTg to thalamic and brainstem nuclei. In addition to participating in motor functions, the PPTg participates in arousal, attention, action selection and reward mechanisms. Finally, we discuss the possibility that the PPTg may be involved in excitotoxic degeneration of the dopaminergic neurons of the substantia nigra through the glutamatergic monosynaptic input that it provides to these neurons.

Funnel plots and choropleth maps in cancer risk communication: a Delphi study.

BACKGROUND: In the last decades, the issues related to health risk communication to stakeholders and citizens involving health care practitioners and local political authorities have been increasingly debated. The study evaluated an alternative strategy to communicate cancer risk to local communities, involving an expert panel of public health operators in comparing two different graphic tools, Funnel Plot and Choropleth map. STUDY DESIGN: A Delphi method process was implemented to achieve a unified consensus on an expert panel of public health operators with regard to weaknesses and strengths of the Funnel Plot and the Choropleth map as tools for cancer risk communication to local communities and other stakeholders. METHODS: Participants were asked to score the efficacy of the two tools using a scale. Six properties were explored through two consecutive consensus rounds. Scales were used to calculate frequencies and the content validity ratio for each domain within the consensus rounds. RESULTS: After the two consecutive rounds, participants expressed their preference in favour of the Choropleth map for its ability to define the spatial location of the risk and to locate any potential cluster, while reaching a consensus with regard to the Funnel Plot properties to identify hot spots, displaying the scope of the phenomenon under investigation, and to show the precision of estimates and communicating the significance of estimates. CONCLUSIONS: The Delphi process allowed us to conclude that Funnel Plot could be used as a complement to the current and commonly used graphical and visual formats to effectively communicate cancer epidemiological data to communities and local authorities, representing a useful tool for empowering the general population.

How should vaccination services be planned, organized, and managed? Results from a survey on the Italian vaccination services.

BACKGROUND: Quality improvement is an increasingly recognized approach to maximize service effectiveness and minimize costs in public health. However, the Italian law never provided for the institutional accreditation of vaccination services. Furthermore, a recently approved law added six more compulsory vaccinations to the original four, which has led to a considerable increase in vaccination efforts, without any previous resources evaluation. The aim of the study was to investigate structural, organizational and managerial characteristics of the Italian vaccination services, in order to suggest the adoption of adequate quality standards. STUDY DESIGN: A survey involving the representatives of the Italian Regions and Autonomous Provinces was performed between September 2017 and September 2018. METHODS: An online questionnaire, including 26 items, designed to evaluate the structural, organizational and managerial characteristics of vaccination services was administered. The correlation between the number of vaccination centres and the coverage for each region was used to evaluate the performance of the vaccination services. RESULTS: Respondents from seven Regions, totaling >15,000,000 inhabitants, answered the questionnaire. Overall, each vaccination service was potentially accessed by an average of 519 children aged zero to 24 months, with a ß-coefficient of -0.87 (p = 0.01) for infant vaccination coverage in 2016. Eighty-five percent of vaccination services were provided with architectural features to accommodate the disabled but only 49% provided reserved parking lots. An average of 0.4 physicians and 0.6 other healthcare workers per 10,000 inhabitants were employed in vaccination services, with complete computerization in 74% of them. CONCLUSION: The inverse relation between vaccination services' spatial accessibility and vaccination coverage suggests that distance and accessibility of vaccination services should be considered in planning. This survey constitutes a baseline data for Italian vaccination services that could be useful for decision makers in establishing minimum requirements to provide high-quality preventive healthcare service.

Effectiveness of an educational intervention on seasonal influenza vaccination campaign adherence among healthcare workers of the Palermo University Hospital, Italy.

INTRODUCTION: Healthcare workers are continuously exposed to the risk of being infected by influenza viruses during their work, thus representing a threat especially for fragile patients. Although the Italian Ministry of Health strongly recommends influenza vaccination for all HCWs, coverage levels in Italy are still far from the expected. Several studies report that one of the preferred strategies to improve vaccination coverage among Healthcare Workers is improving vaccination knowledge through specific multidisciplinary courses. To assess the effectiveness of an educational intervention on influenza vaccination coverage among Healthcare Workers a study was conducted at "Paolo Giaccone" University Hospital of Palermo, in the occasion of the 2016/2017 seasonal influenza vaccination campaign. MATERIAL AND METHODS: Educational interventions on influenza infection and vaccination were organized involving personnel of the hospital units in which patients were more fragile. The Healthcare Workers who volunteered attend the course were considered as the intervention group, while two controls for each case, composed by Healthcare Workers not attending it, were randomly selected from the same unit. For both groups, a questionnaire was used to investigate attitude and behaviors toward influenza vaccination, while vaccination coverage data were obtained throughout the Hospital informational data system. RESULTS: Overall, out of the 125 participants, 38 (30.4%) followed the course (intervention group) and 87 (69.6%), not attending the course, represented the control group; later, only 43 Healthcare Workers out of 125 (34.4%) underwent vaccination during the season considered. In particular, after the educational intervention, 42% of the attending Healthcare Workers got vaccinated, while vaccination prevalence in the control group was 31%. The Healthcare Workers who underwent vaccination reported, before the intervention, a higher risk perception for contracting (transmitting) influenza compared to those not vaccinated (p<0.05), while no significant difference in risk perception of transmitting influenza to their patients was reported between the two groups. DISCUSSION: Despite the training provided, and an improvement in vaccination adherence by the Healthcare Workers involved, coverage obtained was lower than recommended to reduce influenza spread in hospital contexts. In conclusion, our data suggest that specific training alone may play a role in the improvement of influenza vaccination adherence, but it should be integrated by a wider range of public health measures, including mandatory vaccination.

Effect of fish length and nutritional condition on the fecundity of distressed Atlantic cod Gadus morhua from the Baltic Sea.

The disappearance of larger individuals and the decrease in individual body condition suffered by Atlantic cod Gadus morhua in the eastern Baltic during the past two decades can be expected to affect the stock reproductive output. To investigate this, female G. morhua were collected during the spawning and pre-spawning period in 2015-2016. The current individual potential fecundity (FP ) of eastern Baltic G. morhua was estimated and analysed in relation to total length (LT ) and indices of nutritional status such as body condition (K) and hepato-somatic index (IH ) using generalized linear models. In addition, the current prevalence of atresia and its potential relation to K were investigated. Moreover, a calibration curve to estimate FP from oocyte diameter, based on the autodiametric oocyte counting method, was established for the first time for eastern Baltic G. morhua and can be used for future fecundity studies on this stock. The results showed that FP was mainly positively related to fish length, but K and IH also contributed significantly to the variation in FP . The model predicted that fish with K = 1·2 have a FP 51% higher than fish of the same LT with K = 0·8. The prevalence of fecundity regulation by atresia was 5·8%, but it was found only in fish in the pre-spawning maturity stage and with low K. Temporal changes in biological features such as the length composition and individual body condition of eastern Baltic G. morhua, should be accounted for when estimating stock reproductive potential.

Improvement in vaccination knowledge among health students following an integrated extra curricular intervention, an explorative study in the University of Palermo.

INTRODUCTION: Vaccination coverages threaten to decrease because of false beliefs in their unsafety and inefficacy. Therefore formation of future health-care workers on this topic is fundamental to deal with any doubt and to promote active immunization among general population. METHODS: In order to assess health-care students' knowledge about vaccination before an integrated seminar on this topic, and to evaluate their improvement after the educational intervention, an integrated educational intervention was held by a multidisciplinary team. Before and after the seminar, 118 students of medicine and biology schools at Palermo University were asked to answer 10 multiple-choice questions regarding vaccine history, mechanism of action, side effects, composition, use and nowadays issues (hesitancy). Two more questions investigating possible changes on students' attitudes towards vaccination and the usefulness of the formative intervention, were added at the post-test phase of the survey. RESULTS: Eighty-one out of 118 students (68.6%) answered to both pre- and post-test questions. 97.6% and 81.5% of the participating group also completed the two additional questions about their improvement in knowledge (question 11) and attitudes (question 12) towards vaccinations. The post-test results showed a significant improvement for all questions administered, except for number 3 (about a specific immunological content), with an overall percentage of correct answers increasing from 38.8% to 77.6% (p©< 0.001). CONCLUSIONS: The present explorative study put the basis for future studies, stronger in the methodology, and highlights the importance of educating health-care professions students by integrated extra-curricular intervention to be held early in their degree curricula and in order to improve knowledge and attitudes towards vaccinations and to prepare them to promote vaccines among the general population.

PKCε is a regulator of hypertrophic differentiation of chondrocytes in osteoarthritis.

OBJECTIVE: Osteoarthritis (OA) is a common and highly debilitating degenerative disease whose complex pathogenesis and the multiplicity of the molecular processes involved, hinder its complete understanding. Protein Kinase C (PKC) novel isozyme PKCε recently proved to be an interesting molecule for further investigations as it can represent an intriguing, new actor in the acquisition of a OA phenotype by the chondrocyte. DESIGN: PKCε was modulated in primary chondrocytes from human OA patient knee cartilage samples by means of short hairpin RNA (ShRNA) and the expression of cartilage specific markers observed at mRNA and protein level. The involvement of Histone deacetylases (HDACs) signaling pathway was also investigated through the use of specific inhibitors MS-275 and Inhibitor VIII. RESULTS: PKCε loss induces up-regulation of Runt-domain transcription factor (RUNX2), Metalloproteinase 13 (MMP13) and Collagen X (COL10) as well as an enhanced calcium deposition in OA chondrocyte cultures. In parallel, PKCε knock-down also leads to SOX9 and Collagen II (COL2) down-modulation and to a lower deposition of glycosaminoglycans (GAGs) in the extracellular matrix (ECM). This novel regulatory role of PKCε over cartilage hypertrophic phenotype is exerted via an HDAC-mediated pathway, as HDAC2 and HDAC4 expression is modulated by PKCε. HDAC2 and HDAC4, in turn, are at least in part responsible for the modulation of the master transcription factors RUNX2 and SOX9, key regulators of chondrocyte phenotype. CONCLUSIONS: PKCε prevents the phenotypic progression of the OA chondrocyte, acting on cartilage specific markers through the modulation of the transcription factors SOX9 and RUNX2. The loss of PKCε enhances, in fact, the OA hypertrophic phenotype, with clear implications in the pathophysiology of the disease.

Influenza vaccination in high-risk groups: a revision of existing guidelines and rationale for an evidence-based preventive strategy.

Influenza, an infectious respiratory disease, is one of the main causes of excess winter deaths (EWDs) in Europe. Annual flu epidemics are associated with high morbidity and mortality rates, especially among the elderly, those with underlying health conditions and pregnant women. Health Care Workers (HCWs) are also considered at high risk of both contracting influenza and spreading the virus to vulnerable patients. During the 2014/2015 season, the excess winter mortality rates observed in countries of the northern hemisphere (EuroMOMO network) and in Italy (+13%) were strongly related to the intensity of influenza circulation. Influenza vaccination is the most important public health intervention to prevent seasonal influenza transmission and infection. However, to date, influenza vaccination coverage reported in Europe (including high-risk groups) is still largely unsatisfactory. This study analyzes some international and European guidelines on influenza vaccination and the rationale that underlies evidence- based public health intervention for the prevention of influenza among the principal high-risk groups: a) the elderly (subjects aged 65 years or older); b) subjects with underlying health conditions; c) pregnant women; d) healthcare workers. Only by achievement recommended influenza vaccination coverage among high-risk groups in all European countries can we reduce the burden of disease.

Post-vaccine measles in a child with concomitant influenza, Sicily, Italy, March 2015.

We describe the occurrence of measles in an 18 month-old patient in Sicily, Italy, in March 2015, who received the first dose of a measles-containing vaccine seven days before onset of prodromal symptoms. Measles virus infection was confirmed by PCR and detection of specific immunoglobulin; viral genotyping permitted the confirmation of a vaccine-associated illness. The patient had a concurrent influenza virus infection, during a seasonal epidemic outbreak of influenza.

Allele frequencies of microsatellite loci for genetic characterization of a Sicilian bovine population.

Short tandem repeats are used as an effective method to trace DNA markers in genotyping. Using a standardized kit, we tested 11 microsatellite markers recommended by the International Society for Animal Genetics (ISAG) in a sample of 495 Sicilian cattle. The aim of this study was to investigate the allele frequencies in the Sicilian cattle population to provide a reference database and at the same time to assess the use of the ISAG microsatellite panel for pedigree analysis. DNA samples were collected from blood and amplified in an 11-plex polymerase chain reaction (PCR); PCR products were injected in a 3130 Genetic Analyzer. All loci showed high mean polymorphism information content (0.768), and the observed mean heterozygosity was less than the expected value (0.732 vs 0.794, respectively). The exact test for Hardy-Weinberg proportions, allele number, and inbreeding coefficient were calculated. Our results indicated that equilibrium was not always maintained. The observed mean homozygote value exceeded the expected value (132.81 vs 102.14), but no evidence for allele dropout was found. These results could be explained by a non-random mating; further studies using a larger number of animals could confirm or invalidate this hypothesis. The probability of identity and exclusion of a locus were also estimated and proved to be useful in paternity testing. The ISAG microsatellite panel is useful to screen the Sicilian bovine kinship. Currently, an allele frequency database is being constructed.

Genomic and epidemiological evidence for the emergence of a L. infantum/L. donovani hybrid with unusual epidemiology in northern Italy.

Leishmania (L.) infantum is one of the main causative agents of animal and human leishmaniasis across many endemic areas in South America, Europe, North Africa, and Asia. Despite its clinical significance, little is known about the genetic diversity of L. infantum circulating in a given endemic area. Here, we investigate this important open question by applying a comparative genomics approach to seven L. infantum isolates from different hosts and Italian regions, including the northern part of the country (Emilia-Romagna, RER), Sicily, and Sardinia, as an initial attempt to explore the breadth of parasite genetic heterogeneity in Italy. Additionally, microsatellite analysis was carried out to compare the isolates from RER with other 70 L. infantum strains from the same region as well as 65 strains belonging to the L. donovani complex from other countries. We revealed important karyotypic instability and identified strain-specific changes in gene dosage, which affected important virulence factors such as amastins and surface antigen-like proteins. Single nucleotide polymorphism-based clustering analysis of these genomes together with over 80 publicly available L. infantum and L. donovani genomes placed the Italian isolates into three geographically distinct clusters within the Mediterranean basin and uncovered three isolates clustering with putative L. infantum/L. donovani hybrids isolated in Cyprus. As judged by microsatellite profiling, these hybrid isolates are representative of a sub-population of parasites circulating in northern Italy that preferentially infect humans but not dogs. Our results place Italy at the crossroads of L. infantum infection in the Mediterranean and call attention to the public health risk represented by the introduction of non-European Leishmania species.IMPORTANCEThis study closes important knowledge gaps with respect to Leishmania (L.) infantum genetic heterogeneity in a given endemic country, as exemplified here for Italy, and reveals genetic hybridization as a main cause for re-emerging human leishmaniasis in northern Italy. The observed high diversity of Leishmania parasites on the Italian peninsula suggests different geographical origins, with genomic adaptation to various ecologies affecting both pathogenicity and transmission potential. This is documented by the discovery of a putative L. infantum/L. donovani hybrid strain, which has been shown to preferentially infect humans but not dogs. Our results provide important information to health authorities, which need to consider the public health risk represented by the introduction of new Leishmania species into EU countries due to population displacement or travel from countries where exotic/allochthonous parasite species are endemic.

[A full economic evaluation of extensive vaccination against rotavirus with RIX4414 vaccine at National and Regional level in Italy]. / Una valutazione economica completa della vaccinazione estensiva contro i rotavirus con il vaccino RIX4414 a livello nazione e regionale in Italia.

Vaccination of all healthy children against rotavirus (RV) has been recommended, since the availability of vaccines, both in Europe (PIDJ) and Italy (pediatricians). The aims of universal vaccination against RV include the protection of children against moderate/severe gastroenteritis forms by RV (GARV), prevent hospitalizations, reduce the severity and duration of the disease, and reduce morbidity and socioeconomic costs. Payers need to informed regarding the efficacy and the healthcare utilization related to RV vaccination in order to decide in favour of its extensive implementation. The aim of this paper is to assess the clinical and financial impact of the extensive vaccination aganist RV both at National and Regional level. Particular attention, compared to the previous analysis (Standaert et al, 2008) has been given to the influence of herd immunity (HI) on cost-utility results of vaccination against-RV. Methods. The analysis was conducted with the Markovian model previously used by Standaert B et al and updated for comparing costs and benefits associated with a situation of vaccination anti-RV that includes efficacy data due to HI, with a situation without vaccination. For the base case is assumed an annual coverage of 90%, where the effect of HI is present in the population at risk (0-5 years) and extended to children who have not been vaccinated, adding as conservative assumption, a further 10% to the efficacy of the vaccine, compared to 15% determined by several published studies. Two analysis have been made based on this model: a cost-utility analysis that compared vaccination with two doses of RIX441410 administered at 2 and 3 months after birth compared with no vaccination from National Health Service and Society perspective; a budget impact analysis at National and Regional level. The evaluation has as its main element the reduction of cases of infection through universal vaccination and consequent reduction of Garv events and nosocomial infections. Results. From the NHS perspective, in a cohort of 555,791 born in Italy in 2011, the annual number of hospitalizations due to RV infections in the absence of vaccination is estimated to be 14,550 units. Assuming that 90% of newborns receive two doses of the vaccine, and including an additional effect of HI to the efficacy of the vaccine, vaccination would lead to a reduction of 71% of cases of Garv (176,804 cases in less) and a 86% of hospitalizations due to Garv (12,913 fewer cases), with an impact on quality of life and mortality as a consequence of vaccination. The introduction of the vaccine would lead to a gain of 0.0014 QALYs and 0.0022 life-years gained per child compared to a situation without vaccination (assuming a discount rate of 3% on future benefits). The reduction of GARV also would lead to a strong economic impact. The introduction of the vaccine would lead to a saving of € 25.41 per child or a saving of more than € 14 million for the whole population included in the analysis. Cost reduction increase significantly from the perspective of society and introducing the indirect costs due to lost productivity. In this case, the savings due to the introduction of vaccination would increase to € 67,747,654 in the total cohort, or € 121.89 per child. In an alternative scenario, where HI is excluded, RIX4414 remains dominant (0.0013 QALYs gained and € 22.14 per child saved). The budget impact analysis shows that, as early as the second year, the additional cost of the vaccine is more than offset by a reduction in costs of the disease, which leads to savings for the NHS, which increases from year 3. In a time horizon of 5 years (without the discount rate), the savings for the NHS amount to € 34,440,314. These savings would amount to a cost reduction of € 4.64 per child over 5 years (€ 0.93 per year). The savings due to the introduction of the vaccine were mainly due to a reduction in costs associated with hospitalizations. The budget impact analysis at regional level, has taken a vaccine cost of € 30.00 per dose. Cases of diarrhoea before after vaccination are reduced in each region, based on the number of births, ranging from a minimum of 399 cases avoided for Valle d'Aosta to a maximum of 31,116 cases avoided in Lombardy. In a similar way, the number of hospitalizations due to GARV are reduced considerably, from a minimum of 36 cases in Valle d'Aosta to a maximum of 3,096 in Lombardy. Obviously, these reductions are greater in regions with 30,000 or more births per year. Conclusions. This study suggests that a universal vaccination anti-RV with 2 doses of RIX4414 brings significant clinical and economic benefits both at National and Regional level. The indirect effects of the vaccine (HI) could generate protection even in unvaccinated children with health gain and a number of cases by GARV much less than those that would vaccinating small groups of children and with a cost of illness, for NHS, which would be reduced significantly, despite the additional costs of the vaccine as early as the second year of vaccination. Productivity losses due to absence from work of a parent, as well as all other costs included in the model, show that is precisely the society to pay the consequences, from economic and social point of view. Considering the citizen in the role of private payer, we must stress as for him, the savings generated by vaccination, whether universal or with demand for cost-sharing by the health service, prove significant with a major health gain for the population under study.

Risk assessment in ginecology and obstetrics in Sicily: an approach based on Wolff's Criteria.

OBJECTIVES: To apply Wolff's Criteria to hospital discharge records (HDR) in order to detect adverse events worthy of further study. METHODS: Gynecology and Obstetrics Units of three Sicilian hospitals were considered and HDR regarding ordinary and day hospital admissions in 2008 were collected. A matched case-control study was designed, by random selection of 10 controls at maximum for each case. Matching was performed on the variables age and speciality of admission (gynecology or obstetrics). RESULTS: Out of a total of 7011 HDR examined, 114 cases were identified with Wolff's Criteria. Multivariate analysis confirmed a statistically significant association with the origin of admission, diagnosis at the acceptance and length of stay: there was a decreased risk of Wolff's event in patients having urgent admission compared to elective (OR = 0.47, 95% CI = [0.28-0.78]), an increased risk in patients reporting tumor (OR = 5:41, 95 % CI [1.89-15.47]) and other causes (OR = 2.16, 95% CI [1.10-4.24]) compared to delivery diagnosis at acceptance and in patients whose length of stay was more than 6 days (OR = 23.17, 95% CI = [12.56-42.7]) compared to less or equal than 3 days CONCLUSION: Wolff's Criteria can be applied for the analysis of clinical risk in hospitals with different structural characteristics, on condition that the HDR database is complete and good quality.

Use of mesenchymal stem cells to enhance or restore fertility potential: a systematic review of available experimental strategies.

STUDY QUESTION: To what extent does regenerative medicine with stem cell therapy help to address infertility issues for future clinical application? SUMMARY ANSWER: Regenerative medicine using different stem cell sources is yielding promising results in terms of protecting the ovarian reserve from damage and senescence, and improving fertility potential in various preclinical settings. WHAT IS KNOWN ALREADY: Regenerative medicine using stem cell therapy is emerging as a potential strategy to address a number of issues in the field of human reproduction. Indeed, different types of adult and fetal mesenchymal stem cells (MSCs) have been tested with promising results, owing to their ability to differentiate into different tissue lineages, move toward specific injured sites (homing), and generate a secretome with wound-healing, proangiogenic, and antioxidant capacities. STUDY DESIGN SIZE DURATION: Guided by the checklist for preferred reporting items for systematic reviews and meta-analyses, we retrieved relevant studies from PubMed, Medline, and Embase databases until June 2023 using the following keywords: 'mesenchymal stem cells' AND 'ovarian follicles' OR 'ovarian tissue culture' OR 'ovarian follicle culture' OR 'cumulus oocyte complex'. Only peer-reviewed published articles written in English were included. PARTICIPANTS/MATERIALS SETTING METHODS: The primary outcome for the experimental strategies was evaluation of the ovarian reserve, with a focus on follicle survival, number, and growth. Secondary outcomes involved analyses of other parameters associated with the follicle pool, such as hormones and growth factors, ovarian tissue viability markers including oxidative stress levels, oocyte growth and maturation rates, and of course pregnancy outcomes. MAIN RESULTS AND THE ROLE OF CHANCE: Preclinical studies exploring MSCs from different animal origins and tissue sources in specific conditions were selected (n = 112), including: in vitro culture of granulosa cells, ovarian tissue and isolated ovarian follicles; ovarian tissue transplantation; and systemic or intraovarian injection after gonadotoxic or age-related follicle pool decline. Protecting the ovarian reserve from aging and gonadotoxic damage has been widely tested in vitro and in vivo using murine models and is now yielding initial data in the first ever case series of patients with premature ovarian insufficiency. Use of MSCs as feeder cells in ovarian tissue culture was found to improve follicle outcomes and oocyte competence, bringing us one step closer to future clinical application. MSCs also have proved effective at boosting revascularization in the transplantation site when grafting ovarian tissue in experimental animal models. LIMITATIONS REASONS FOR CAUTION: While preclinical results look promising in terms of protecting the ovarian reserve in different experimental models (especially those in vitro using various mammal experimental models and in vivo using murine models), there is still a lot of work to do before this approach can be considered safe and successfully implemented in a clinical setting. WIDER IMPLICATIONS OF THE FINDINGS: All gathered data on the one hand show that regenerative medicine techniques are quickly gaining ground among innovative techniques being developed for future clinical application in the field of reproductive medicine. After proving MSC effectiveness in preclinical settings, there is still a lot of work to do before MSCs can be safely and effectively used in different clinical applications. STUDY FUNDING/COMPETING INTERESTS: This study was supported by grants from the Fonds National de la Recherche Scientifique de Belgique (FNRS-PDR T.0077.14, FNRS-CDR J.0063.20, and grant 5/4/150/5 awarded to Marie-Madeleine Dolmans), Fonds Spéciaux de Recherche, and the Fondation St Luc. None of the authors have any competing interest to disclose. REGISTRATION NUMBER: N/A.