Risk Factors for Lung Cancer in the Province of Lecce: Results from the PROTOS Case-Control Study in Salento (Southern Italy).

In the province of Lecce (southern Italy), a higher incidence of lung cancer (LC) among men compared to regional and national data was reported. In a sub-area in the center of the province (cluster area), the incidence and mortality for LC was even higher. PROTOS is a case-control study aimed at investigating possible risk factors for LC in the province area. A total of 442 patients with LC and 1326 controls matched by sex and age living in the province of Lecce for at least 10 years were enrolled and georeferenced; they filled in a questionnaire with their personal information and exposures. For each risk factor, an Odds Ratio adjusted for all the other variables was calculated. The risk of LC increased with excessive use of alcohol in women, for those subjects with a family cancer history, for each increase in pack/year of cigarettes, for men more exposed considering the industrial district in the cluster area, and for those using pesticides in agriculture without wearing personal protective equipment. The higher incidence of adenocarcinoma in both sexes suggests that, in addition to cigarette smoking, concurrent exposures to other environmental, occupational, and life-style factors may play a role in increased cancer risk and should be more deeply explored.

Buccal micronucleus cytome assay in children living in an area with low anthropogenic pressure: The EFFE.BI.P. study.

The study aimed to evaluate the micronucleus (MN) frequency in exfoliated buccal cells (EBCs) of 256 6-8-years-old schoolchildren living in a rural area of Salento peninsula (Southern Italy) with low anthropogenic pressure and with a normal rate of chronic diseases in order to determine the basal level of MN and identify which factors are able to influence it. Information about the personal data, lifestyles and dietary habits of the children were obtained by the administration of a questionnaire to their parents. The buccal micronucleus cytome assay was performed to evaluate the presence of early genotoxic effects among the children. In addition, the level of environmental exposure was assessed by sampling atmospheric particulate fractions near the schools attended by participants. The association between MN frequency and individual or environmental factors was also assessed. The children had a mean MN frequency of 0.27 ± 0.43‰ (95%CI = 0.22-0.33). This frequency was positively associated with vehicular traffic (OR = 2.99; 95%CI = 1.15-7.74) and negatively associated with a high educational level of the mother (OR = 0.41; 95%CI = 0.18-0.95) and physical exercise (OR = 0.56; 95%CI = 0.32-0.57). Data on genotoxic effects in buccal cells found in this study could be considered as the MN level in a pediatric population not exposed to environmental pollution.

The expression of microRNAs and exposure to environmental contaminants related to human health: a review.

Environmental contaminants exposure may lead to detrimental changes to the microRNAs (miRNAs) expression resulting in several health effects. miRNAs, small non-coding RNAs that regulate gene expression, have multiple transcript targets and thereby regulate several signalling molecules. Even a minor alteration in the abundance of one miRNA can have deep effects on global gene expression. Altered patterns of miRNAs can be responsible for changes linked to various health outcomes, suggesting that specific miRNAs are activated in pathophysiological processes. In this review, we provide an overview of studies investigating the impact of air pollution, organic chemicals, and heavy metals on miRNA expression and the potential biologic effects on humans.Abbreviations: AHRR, aryl-hydrocarbon receptor repressor; AHR, aryl-hydrocarbon receptor; As, arsenic; BCL2, B-cell lymphoma 2; BCL2L11, B-cell lymphoma 2 like 11; BCL6, B-cell lymphoma 6; BPA, bisphenol A; CVD, cardiovascular diseases; CD40, cluster of differentiation 40; CCND1, Cyclin D1; CDKN1A, cyclin-dependent kinase inhibitor 1A; Cr, chromium; CTBP1, C-terminal binding protein 1; CXCL12, C-X-C motif chemokine ligand 12; DAZAP1, deleted in azoospermia associated protein 1; DEP, diesel exhaust particles; EGFR, epidermal growth factor receptor; eNOS, endothelial nitric oxide synthase; EVs, extracellular vesicles; FAK, focal adhesion kinase; FAS, fas cell surface death receptor; FOXO, forkhead box O; HbA1c, glycated hemoglobin; Hg, mercury; HLA-A, human leukocyte antigen A; HMGB, high-mobility group protein B; IFNAR2, interferon alpha receptor subunit 2; IL-6, interleukin-6; IRAK1, interleukin 1 receptor associated kinase 1; JAK/STAT, janus kinase/signal transducers and activators of transcription; MAPK, mitogen-activated protein kinase; miRNAs, microRNAs; MVs, microvesicles; NCDs, noncommunicable diseases; NFAT, nuclear factor of activated T cells; NFkB, nuclear factor kappa B; NRF2, nuclear factor, erythroid-derived 2; NRG3, neuregulin 3; O3, ozone; OP, organophosphorus pesticides; PAHs, polycyclic aromatic hydrocarbons; Pb, lead; PCBs, polychlorinated biphenyls; PDCD4, programmed cell death 4; PDGFB, platelet derived growth factor subunit beta; PDGFR, platelet-derived growth factor receptor; PI3K/Akt, phosphoinositide-3-kinase/protein kinase B; PKA, protein kinase A; PM, particulate matter; PRKCQ, protein kinase C theta; PTEN, phosphatase and tensin homolog; SORT1, sortilin 1; TGFß, transforming growth factor-ß; TLR, toll-like receptor; TNF, tumor necrosis factors; TRAF1, tumor necrosis factors-receptor associated factors 1; TRAP, traffic-related air pollution; TREM1, triggering receptor expressed on myeloid cells 1; TRIAP1, TP53 regulated inhibitor of apoptosis 1; VCAM-1, vascular cell adhesion molecule 1; VEGFA, vascular endothelial growth factor A; XRCC2, X-ray repair cross complementing 2; YBX2, Y-box-binding protein 2; ZEB1, zinc finger E-box-binding homeobox 1; ZEB2, zinc finger E-box-binding homeobox 2; 8-OH-dG, 8-hydroxy-guanine.

The Water Safety Plan Approach: Application to Small Drinking-Water Systems-Case Studies in Salento (South Italy).

BACKGROUND: The quality of water for human consumption is an objective of fundamental importance for the defense of public health. Since the management of networks involves many problems of control and efficiency of distribution, the Water Safety Plan (WSP) was introduced to address these growing problems. METHODS: WSP was applied to three companies in which the water resource assumes central importance: five water kiosks, a third-range vegetable processing company, and a residence and care institution. In drafting the plan, the terms and procedures designed and tested for the management of urban distribution systems were applied to safeguard the resource over time. RESULTS: The case studies demonstrated the reliability of the application of the model even to small drinking-water systems, even though it involved a greater effort in analyzing the incoming water, the local intended use, and the possibilities for managing the containment of the dangers to which it is exposed. This approach demonstrates concrete effectiveness in identifying and mitigating the dangers of altering the quality of water. CONCLUSIONS: Thanks to the WSP applied to small drinking-water systems, we can move from management that is focused mainly on verifying the conformity of the finished product to the creation of a global risk assessment and management system that covers the entire water supply chain.

Early-Life Exposure to Environmental Air Pollution and Autism Spectrum Disorder: A Review of Available Evidence.

The number of children diagnosed with Autism Spectrum Disorder (ASD) has rapidly increased globally. Genetic and environmental factors both contribute to the development of ASD. Several studies showed linkage between prenatal, early postnatal air pollution exposure and the risk of developing ASD. We reviewed the available literature concerning the relationship between early-life exposure to air pollutants and ASD onset in childhood. We searched on Medline and Scopus for cohort or case-control studies published in English from 1977 to 2020. A total of 20 articles were selected for the review. We found a strong association between maternal exposure to particulate matter (PM) during pregnancy or in the first years of the children's life and the risk of the ASD. This association was found to be stronger with PM2.5 and less evident with the other pollutants. Current evidence suggest that pregnancy is the period in which exposure to environmental pollutants seems to be most impactful concerning the onset of ASD in children. Air pollution should be considered among the emerging risk factors for ASD. Further epidemiological and toxicological studies should address molecular pathways involved in the development of ASD and determine specific cause-effect associations.

The influence of lifestyle factors on miRNA expression and signal pathways: a review.

The term 'lifestyle' includes different factors that contribute to the maintenance of a good health status. Increasing evidences suggest that lifestyle factors may influence epigenetic mechanisms, such as miRNAs expression. The dysregulation of miRNAs can modify the expression of genes and molecular pathways that may lead to functional alterations. This review summarizes human studies highlighting that diet, physical activity, smoking and alcohol consumption may affect the miRNA machinery and several biological functions. Most miRNAs are involved in molecular pathways that influence inflammation, cell cycle regulation and carcinogenesis resulting in the onset or progression of pathological conditions. Investigating these interactions will be pivotal for understanding the etiology of pathologic processes, the potential new treatment strategies and for preventing diseases.

Water Quality Assessment: A Quali-Quantitative Method for Evaluation of Environmental Pressures Potentially Impacting on Groundwater, Developed under the M.I.N.O.Re. Project.

Background: At global level, the vulnerability of aquifers is deteriorating at an alarming rate due to environmental pollution and intensive human activities. In this context, Local Health Authority ASL Lecce has launched the M.I.N.O.Re. (Not Compulsory Water Monitoring Activities at Regional level) project, in order to assess the vulnerability of the aquifer in Salento area (Puglia Region) by performing several non-compulsory analyses on groundwater samples. This first paper describes the quali-quantitative approach adopted under the M.I.N.O.Re. project for the assessment of environmental pressures suffered by groundwater and determines the number of wells to be monitored in specific sampling areas on the basis of the local potential contamination and vulnerability of the aquifer. Methods: We created a map of the entire Lecce province, interpolating it with a grid that led to the subdivision of the study area in 32 quadrangular blocks measuring 10 km × 10 km. Based on current hydrogeological knowledge and institutional data, we used GIS techniques to represent on these 32 blocks the 12 different layers corresponding to the main anthropic or environmental type of pressures potentially impacting on the aquifer. To each kind of pressure, a score from 0 to 1 was attributed on the basis of the potential impact on groundwater. A total score was assigned to each of the 32 blocks. A higher number of wells was selected to be monitored in those blocks presenting higher risk scores for possible groundwater contamination due to anthropic/environmental pressures. Results: The range of total scores varied from 2.4 to 42.5. On the basis of total scores, the 10 km × 10 km blocks were divided into four classes of environmental pressure (1st class: from 0,1 to 10,00; 2nd class: from 10,01 to 20,00; 3rd class: from 20,1 to 30,00; 4th class: from 30,01 to 42,50). There were 11 areas in the 1st class, 9 areas in the 2nd class, 8 areas in the 3rd class and 4 areas in the 4th class. We assigned 1 monitoring well in 1st class areas, 2 monitoring wells in 2nd class areas, 3 monitoring wells in 3rd class areas and 4 monitoring wells in 4th class areas. Conclusion: The methodology developed under the M.I.N.O.Re. project could represent a useful model to be used in other areas to assess the environmental pressures suffered by aquifers and the quality of the groundwater.

Micronucleus Frequency in Exfoliated Buccal Cells of Children Living in an Industrialized Area of Apulia (Italy).

Micronuclei (MN) are biomarkers of early biological effect often used for detecting DNA damage in human population exposed to genotoxic agents. The aim of this study was to evaluate the frequency of MN in exfoliated buccal cells of children living in an industrialized (impacted) area compared with that found in children living in a control area without significant anthropogenic impacts. A total of 462 6-8-year-old children (206 in the impacted area, 256 in the control area) attending primary school were enrolled. A questionnaire was administered to the parents of the recruited children to obtain information about personal data, lifestyles, and food habits of their children. Atmospheric particulate fractions were collected near the involved schools to assess the level of environmental exposure of the children. The presence of MN was highlighted in 68.4% of children living in the impacted area with a mean MN frequency of 0.66‱ ± 0.61‱. MN positivity and frequency were significantly lower in the control area (37.1% and 0.27‱ ± 0.43‱, respectively). The frequency of MN was positively associated with quasi-ultrafine particulate matter (PM0.5), traffic near the home, and consuming barbecued food; while adherence to the Mediterranean diet and practicing sport were negatively associated.

Adherence to Mediterranean diet of children living in small Southern Italian villages.

A cross-sectional study was conducted in the primary schools of five small villages of Salento Peninsula to evaluate the adherence to the Mediterranean Diet (MD) of 282 6-8-years old children in relation to lifestyles and socio-economic factors. The parents of children completed a self-administered questionnaire to evaluate the prevalence of personal, behavioural and socio-economic factors of their sons. Children's anthropometric measurements were also taken. The adherence to the MD was assessed using the Mediterranean Diet Quality Index for Children and Adolescents (KIDMED). The chi-square test was used to detect any differences among groups of children. Overall, 27.0% of children showed low adherence to the MD (KIDMED ≤3), 59.6% medium adherence (KIDMED 4-7) and 13.5% high adherence (KIDMED ≥8). The adherence to the MD was associated with the educational level (whether graduated or not) of the mother and the occupational status (whether employed or not) of both parents.

Micronuclei in Exfoliated Buccal Cells of Children Living in a Cluster Area of Salento (Southern Italy) with a High Incidence of Lung Cancer: The IMP.AIR Study.

During the years 2014⁻2016 the University of Salento performed the "Impact of Air Quality on Health of Residents in the Municipalities of Cutrofiano, Galatina, Sogliano Cavour, Soleto and Sternatia" (IMP.AIR) study, an epidemiological-molecular research project aiming to evaluate early DNA damage in children living in an area of Salento with high incidence of lung cancer among the male population. One hundred and twenty-two children aged 6⁻8 years attending primary school were enrolled and the frequency of micronucleated cells (MNC) in oral mucosa was evaluated. In addition, a questionnaire was administered to parents to obtain information about personal data, anthropometric characteristics and lifestyles (physical activity, food habits, family context) of the children and perform a multivariate analysis to detect any factors associated with MNC occurrence. Data on airborne pollutants detected in the study area were acquired by the Regional Agency for the Environmental Protection. The presence of MNC was highlighted in about 42% of children with a mean MNC frequency of 0.49‰. The frequency of MNC was associated to obesity, consumption of red or processed meat and having a mother who smokes. Moreover, the prevalence of biomarkers was higher than in another area of Salento not included in the cluster area.

Health Risk Associated with Exposure to PM10 and Benzene in Three Italian Towns.

Air pollution in urban areas is a major concern as it negatively affects the health of a large number of people. The purpose of this study was to assess the inhalation health risk for exposure to PM10 and benzene of the populations living in three Italian cities. Data regarding PM10 and benzene daily measured by "traffic" stations and "background" stations in Torino, Perugia, and Lecce during 2014 and 2015 were compared to the limits indicated in the Directive 2008/50/EC. In addition, an inhalation risk analysis for exposure to benzene was performed for adults and children by applying the standard United States Environmental Protection Agency's (USEPA) methodology. The levels of PM10 detected in Torino exceeded the legal limits in both years with an increased mean concentration >10 µg/m³ comparing with background station. Benzene concentrations never exceeded the legislative target value. The increased cancer risk (ICR) for children exposed to benzene was greater than 1 × 10-6 only in the city of Torino, while for adults, the ICR was higher than 1 × 10-6 in all the cities. The results suggest the need for emission reduction policies to preserve human health from continuous and long exposure to air pollutants. A revision of legal limits would also be recommended.

Heavy Environmental Pressure in Campania and Other Italian Regions: A Short Review of Available Evidence.

The area of Naples and Campania region, in Italy, are experiencing the dramatic consequences of diffuse and illegal waste dumping, resulting in possible threats to human health. This area has been referred to as the "Land of Fires" because of the common practice of waste burning. International interest in the Campania "waste emergency" has triggered several epidemiological studies. This article is aimed at highlighting the body of evidence available concerning human and environmental contamination in the Campania region, and considers the possible lack of comparable knowledge about the situation in other areas suffering from high environmental pollution. We analyzed the results of studies addressing environmental pollution and population health in the Campania region, starting from the most recent reviews on this topic, and compared their findings with those concerning other regions. We reviewed 18 studies of epidemiological/cancer surveillance and human or animal biomonitoring. These studies show worrying results, which could be considered comparable to those available for other Italian areas impacted by heavy industrial activities. The release of environmental contaminants associated with waste incineration and waste disposal in landfills poses a risk to public health, as shown by a number of studies (although not conclusively). The current knowledge available for the Campania region is better than that available for other areas which are facing similar problems due to anthropic activities, including illegal waste trafficking. Thus, Naples and Campania could represent a valuable setting to develop general models for studies of environmental and human contamination.

Has VZV epidemiology changed in Italy? Results of a seroprevalence study.

The aim of the study was to evaluate if and how varicella prevalence has changed in Italy. In particular a seroprevalence study was performed, comparing it to similar surveys conducted in pre-immunization era. During 2013-2014, sera obtained from blood samples taken for diagnostic purposes or routine investigations were collected in collaboration with at least one laboratory/center for each region, following the approval of the Ethics Committee. Data were stratified by sex and age. All samples were processed in a national reference laboratory by an immunoassay with high sensitivity and specificity. Statutory notifications, national hospital discharge database and mortality data related to VZV infection were analyzed as well. A total of 3707 sera were collected and tested. In the studied period both incidence and hospitalization rates decreased and about 5 deaths per year have been registered. The seroprevalence decreased in the first year of life in subjects passively protected by their mother, followed by an increase in the following age classes. The overall antibody prevalence was 84%. The comparison with surveys conducted with the same methodology in 1996-1997 and 2003-2004 showed significant differences in age groups 1-19 y. The study confirms that in Italy VZV infection typically occurs in children. The impact of varicella on Italian population is changing. The comparison between studies performed in different periods shows a significant increase of seropositivity in age class 1 - 4 years, expression of vaccine interventions already adopted in some regions.

Dramatic recovery of steroid-refractory relapsed multiple sclerosis following Fingolimod discontinuation using selective immune adsorption.

BACKGROUND: Selective immune adsorption (SIA) is an emerging method for treating immune-mediated neurological diseases, given its superior safety profile compared to plasma exchange (PEX). However, the available literature concerning Multiple Sclerosis includes no cases of SIA applied to steroid-refractory rebound after Fingolimod discontinuation. CASE PRESENTATION: Here we report the case of a 32-year-old woman suffering from multiple sclerosis treated with Fingolimod and admitted to a Multiple Sclerosis Centre after drug discontinuation due to the occurrence of lymphopenia. During the few weeks preceding admission, the patient experienced progressive and severe neurological deterioration that did not respond to an initial cycle of pulsed high doses of intravenous 6-methyl prednisolone (IVMP). Given the ineffectiveness of a second cycle of IVMP, the patient was treated with plasma immunoadsorption, leading to dramatic functional recovery. The patient then started a neuro-rehabilitation program. About one month after the final SIA procedure the patient started Natalizumab-based therapy, while maintaining a stable neurological condition. We noted significant modification of C3/C4 complement components and total gamma globulin concentrations (IgG) during SIA. CONCLUSIONS: Our observations show that however serious, steroid-refractory neurological deterioration occurring after Fingolimod discontinuation in multiple sclerosis can be treated with selective immune-adsorption therapy which thus represents a good alternative in these cases. It could be speculated that this clinical condition was associated with pattern II of demyelination, given the good response to a form of treatment that acts on autoantibodies. Thus, SIA represented an effective therapeutic strategy for this case of relapsed MS as steroid-resistent rebound post Fingolimod cessation.

Impact of stinging jellyfish proliferations along south Italian coasts: human health hazards, treatment and social costs.

Stinging jellyfish outbreaks represent a health hazard, causing contact dermatitis and systemic reactions. This study investigated the epidemiology, severity, and treatment protocols of jellyfish stings in a coastal area with high tourist development and frequent stinging jellyfish outbreaks of the central Mediterranean (Salento, Southern Italy), and the associated costs for the Italian National Health Service. In 2007-2011, 1,733 bathers (mostly children and females) sought medical assistance following jellyfish stings, the main cause of human pathologies due to contact with marine organisms. The majority of events were reported in the years 2007-2009, whereas the occurrence of cnidarian jellyfish outbreaks has been increasingly reported in the same area since summer 2010. Most symptoms were limited to local and cutaneous reactions; conversely, 8.7% of cases evoked complications, mainly due to allergic reactions. The main drugs used were corticosteroids, locally applied and systemic (46% and 43%, respectively), and with ammonia (74%) as the main non-pharmacological treatment. The estimated cost of jellyfish-related first-aid services along the Salento coastline over the 5-year period was approximately 400,000 Euros. Therefore the management of jellyfish outbreak phenomena need coordinated research efforts towards a better understanding of underlying ecological mechanisms, together with the adoption of effective prevention policy, mitigation strategies, and appropriate planning of health services at tourist hot spots.

[Sero-epidemiology of VZV infection in Italy: impact evaluation of extensive vaccination]. / Sieroepidemiologia dell'infezione da VZV in Italia: valutazione dell'impatto della vaccinazione estensiva.

OBJECTIVE: Eight Italian Regions have begun offering vaccination against varicella to children aged 13-15 months, with a second dose at 5-6 years of age. A serosurvey was conducted to evaluate the impact of extensive vaccination in these "pilot" Regions. DESIGN: Samples were collected in compliance with current legislation and after the approval of the Ethics Committee. The qualitative and quantitative determination of antibodies (IgG) against varicella was performed using an ELISA assay with high sensitivity and specificity. RESULTS: 1,470 samples were analyzed. After the decrease of seropositivity associated with the decay of passively acquired antibody titer, there was a progressive increase of seroprevalence in the other age groups. The comparison with what was recorded in two studies conducted with similar assays in the periods 1996/97 and 2003/04, showed a significant increase in seroprevalence following extensive immunization, particularly in the age classes 1 year, 2-4 years and 5-9 years. The proportion of seropositivity in the age classes 1 year and 2-4 years has more than doubled. CONCLUSIONS: Seroepidemiological studies are a useful tool to evaluate the distribution of susceptible individuals in the population. This study highlighted the impact of extensive vaccination against varicella in eight "pilot" Regions.

Rotavirus Occurrence in Shellfish with Low Levels of E. coli.

The purpose of this study was to evaluate "in field" the accumulation of virus in shellfish and compare it with the concentration of bacterial indicators. Individuals of Mytilus galloprovincialis were placed in two sampling station located in a contaminated coastal bay and in one control station located one kilometer offshore. The presence of Rotavirus and E. coli was assessed weekly both in seawater and in shellfish samples. The Rotavirus genome was detected in water, preliminarily concentrated by tangential flow ultrafiltration method, and in hepatopancreas of mussels by Real-Time PCR. E. coli was enumerated in water matrices by a filtering method and in mussels by the MPN method. Rotaviruses were not recorded in seawater, while in mussels they were detected since third week after placement. E. coli in mussels were always below the limits set in the Regulation (EC) 854/2004. This study suggests the need for a viral indicator to insure the safety for consumption of shellfish.

The clinical potential of blood-proteomics in multiple sclerosis.

BACKGROUND: The aetiology of multiple sclerosis (MS) remains unknown. This hampers molecular diagnosis and the discovery of bio-molecular markers. Consequently, MS diagnostic procedures are complex and criteria for assessing therapeutic efficacy are controversial, suggesting that a pathophysiological rather than an aetiological approach to the disease would be more appropriate. In this regard, blood-proteomics represents a still-unexplored tool. We investigated the potential of proteomics as applied to peripheral blood mononuclear cells (PBMCs) for differentiating treatment-naive RR-MS patients from healthy controls and from IFN-treated RR-MS patients. METHODS: A comparative analysis of PBMC proteins isolated from 13 unselected IFN-treated RR-MS patients, 6 IFN-untreated RR-MS patients and 14 matched healthy controls was performed using two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry. We considered the volume of each spot, expressed as a percentage of the total volume of all spots in the gel. Heuristic clustering was applied to a composite population made up of a random sequence of gels from the different groups in comparison. For the differentially expressed proteins, we applied the Student's t-test to identify only those down- or up-regulated at least 2.5-fold [Ratio(R) ≥ 2.5] with respect to the homologous spots of the compared groups. RESULTS: Rho-GDI2, Rab2 and Cofilin1 were found to be associated with down-regulated and naïve group phenotypes; Cortactin and Fibrinogen beta-Chain Precursor were found to be associated with down-regulated and group-related IFN-treated RR-MS phenotypes. Thus, by means of similarity analysis, the proteomes were homogeneously segregated into three distinct groups corresponding to naive, IFN-treated and healthy control subjects. Interestingly, no separation was found between IFN-treated and healthy controls. Moreover, the molecular phenotypes were consistent with disease pathogenesis. CONCLUSIONS: We demonstrated for the first time, albeit only with preliminary data, the aprioristic possibility of distinguishing naive and IFN-treated MS groups from controls, and naive from IFN-treated MS patients using a blood sample-based methodology (i.e. proteomics) alone. The functional profile of the identified molecules provides new pathophysiological insight into MS. Future development of these techniques could open up novel applications in terms of molecular diagnosis and therapy monitoring in MS patients.

First-time comparison of the in vitro antimalarial activity of Artemisia annua herbal tea and artemisinin.

Artemisia annua tea has been proven to be a very effective treatment for malaria in various clinical trials, but to date its efficacy has not been investigated in vitro. A study was therefore performed to evaluate the effects of A. annua tea on Plasmodium falciparum cultures in vitro. The concentration of artemisinin in the herbal tea preparation was also determined. The herbal tea extract was tested against chloroquine (CQ)-sensitive D10 and CQ-resistant W2 strains of P. falciparum using the parasite lactate dehydrogenase assay. Quantification of artemisinin in the extract of leaves of A. annua was performed using proton nuclear magnetic resonance ((1)H-NMR). Results of the in vitro tests were consistent with the clinical efficacy of A. annua tea [50% inhibitory concentration (IC(50)) for strain D10=1.11±0.21 µg/ml; IC(50) for strain W2=0.88±0.35 µg/ml]. The concentration of artemisinin in A. annua tea (0.18±0.02% of dry weight) was far too low to be responsible for the antimalarial activity. The artemisinin present in the tea is probably co-solubilised with other ingredients, some of which also have antimalarial activity and act synergistically with it. These compounds also merit further research to determine whether their presence hinders the development of parasite resistance compared with pure artemisinin.

Detection of viruses in coastal seawater using Mytilus galloprovincialis as an accumulation matrix.

The purpose of this study was to demonstrate that shellfish can be used to detect enteric viruses in marine waters where they are present at very low concentrations. Aqua-cultured mussels were placed in the sea just off the mouth of a drainage channel affected by human and animal faecal contamination. Samples were taken from the channel, the sea and the mussels at intervals over two 4-week periods. The samples were tested to verify the presence of both rotaviruses and E. coli. Rotaviruses were detected by Real Time-PCR, typed by multiplex PCR and subsequently sequenced. E. coli was enumerated in water matrices by a filtering method and in mussels by the MPN method. The presence of E. coli in the examined matrices demonstrates contamination of faecal origin throughout the studied environments. Rotaviruses were recorded in channel waters, but not in sea water. In both experiments, rotaviruses were detected in mussels 21 and 28 days after being placed in the sea water off the channel mouth. The use of mussels thus enabled the detection of rotaviruses in waters where the high dilution rendered direct investigation impossible. This study indicates that mussels can be used in marine virological surveillance programs.