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ABSTRACTIntratumoral bacteria flexibly contribute to cellular and molecular tumor heterogeneity for supporting cancer recurrence through poorly understood mechanisms. Using spatial metabolomic profiling technologies and 16SrRNA sequencing, we herein report that right-sided colorectal tumors are predominantly populated with Colibactin-producing Escherichia coli (CoPEC) that are locally establishing a high-glycerophospholipid microenvironment with lowered immunogenicity. It coincided with a reduced infiltration of CD8+ T lymphocytes that produce the cytotoxic cytokines IFN-γ where invading bacteria have been geolocated. Mechanistically, the accumulation of lipid droplets in infected cancer cells relied on the production of colibactin as a measure to limit genotoxic stress to some extent. Such heightened phosphatidylcholine remodeling by the enzyme of the Land’s cycle supplied CoPEC-infected cancer cells with sufficient energy for sustaining cell survival in response to chemotherapies. This accords with the lowered overall survival of colorectal patients at stage III-IV who were colonized by CoPEC when compared to patients at stage I-II. Accordingly, the sensitivity of CoPEC-infected cancer cells to chemotherapies was restored upon treatment with an acyl-CoA synthetase inhibitor. By contrast, such metabolic dysregulation leading to chemoresistance was not observed in human colon cancer cells that were infected with the mutant strain that did not produce colibactin (11G5∆ClbQ). This work revealed that CoPEC locally supports an energy trade-off lipid overload within tumors for lowering tumor immunogenicity. This may pave the way for improving chemoresistance and subsequently outcome of CRC patients who are colonized by CoPEC.

Folere (Hibiscus sabdariffa L.) is a traditional plant cultivated in the north of Cameroon near the city of Yagoua. Calyces are used to prepare beverages while the leaves are incorporated into meals. This Cameroonian roselle is characterized by a higher intensity of red color than that found in conspecific samples from Egypt, and it was of interest to us to evaluate its beneficial activities in comparison with European pharmacopoeia standards. The samples were first subjected to phylogenetic analyses. Five samples including samples from Egypt and a white flower variety were subsequently examined according to the European pharmacopeia criteria i.e. (i) taxonomic identification, (ii) sample preparation and characterization (desiccation, TLC, coloring power, presence of foreign matter), and (iii) determination of the acidity index. Anthocyanins and anthocyanidins were obtained using three extracts from methanolic maceration, infusion, and boiling in water. Taken together, the results confirmed that the Cameroonian variety indeed belongs to H. sabdariffa and meets European Pharmacopoeia standards. All extracts contained common anthocyanins (delphinidin-3-sambioside, delphinidin-3-glucoside, cyanidin-3-sambioside, cyanidin-3-glucoside), and delphinidin and cyanidin aglycones were also found in the organic extracts. As a result of determining the amount of anthocyanins by HPLC in Cameroon samples, it was determined that they showed a high concentration of delphinidin type anthocyanins and a concentration 10 times higher than the Egyptian variety. This variety appears to be a very promising candidate for inclusion in industrial preparations, thanks to its high anthocyanin qualitative and quantitative properties, and also maintains economic interest for local populations.

This multicenter study evaluated the IMMY Aspergillus Galactomannan Lateral Flow Assay (LFA) with automated reader for diagnosis of pulmonary aspergillosis in patients with COVID-19-associated acute respiratory failure (ARF) requiring intensive care unit (ICU) admission between 03/2020 and 04/2021. A total of 196 respiratory samples and 148 serum samples (n = 344) from 238 patients were retrospectively included, with a maximum of one of each sample type per patient. Cases were retrospectively classified for COVID-19-associated pulmonary aspergillosis (CAPA) status following the 2020 consensus criteria, with the exclusion of LFA results as a mycological criterion. At the 1.0 cutoff, sensitivity of LFA for CAPA (proven/probable/possible) was 52%, 80% and 81%, and specificity was 98%, 88% and 67%, for bronchoalveolar lavage fluid (BALF), nondirected bronchoalveolar lavage (NBL), and tracheal aspiration (TA), respectively. At the 0.5 manufacturer's cutoff, sensitivity was 72%, 90% and 100%, and specificity was 79%, 83% and 44%, for BALF, NBL and TA, respectively. When combining all respiratory samples, the receiver operating characteristic (ROC) area under the curve (AUC) was 0.823, versus 0.754, 0.890 and 0.814 for BALF, NBL and TA, respectively. Sensitivity and specificity of serum LFA were 20% and 93%, respectively, at the 0.5 ODI cutoff. Overall, the Aspergillus Galactomannan LFA showed good performances for CAPA diagnosis, when used from respiratory samples at the 1.0 cutoff, while sensitivity from serum was limited, linked to weak invasiveness during CAPA. As some false-positive results can occur, isolated results slightly above the recommended cutoff should lead to further mycological investigations.

Alveolar echinococcosis (AE) is a severe liver disease due to infection with the Echinococcus multilocularis larval stage, called the metacestode. Management of AE is based on benzimidazole chemotherapy (albendazole or mebendazole), associated with surgery when possible. Benzimidazoles are the only compounds recommended for the treatment of AE; however, these are parasitostatic, which means that the parasite can resume growth when treatment is interrupted. Also, benzimidazoles can cause liver dysfunction which may prevent their use. Numerous drugs have been reported to have in vitro activity against E. multilocularis, but few had satisfactory in vivo activity, and none were clearly more effective than benzimidazoles. These drugs belong to various therapeutic categories including anti-infective agents (e.g. amphotericin B, mefloquine, pentamidine derivatives), anti-neoplastic compounds (e.g. imatinib, nilotinib, bortezomib), plant-extracted compounds (e.g. thymol, crocin, carvacrol) and others (e.g. metformin, verapamil, thiaclopride). These treatments are generally of limited interest due to their toxicity, their unfavorable pharmacokinetics, or the scarcity of studies involving humans. Apart from benzimidazoles, only amphotericin B, mefloquine and nitazoxanide have been reported to be used for human AE treatment, with unsatisfactory results. Few studies have aimed at developing innovative strategies for AE drug therapy, such as vectorization of drugs using nanoparticles. Altogether, this review emphasizes the urgent need for new therapeutic strategies in AE management, for which there is currently no curative chemotherapy.

Schizophrenia (SCZ) and bipolar disorder (BD) are associated with immunological dysfunctions that have been hypothesized to lead to clinical symptomatology in particular through kynurenine pathway abnormalities. The aim of this study was thus to investigate the impact of serum kynurenine metabolite levels on diagnosis, clinical state, symptom severity and clinical course in a large French transdiagnostic cohort of SCZ and BD patients. Four patient groups (total n = 507) were included in a cross-sectional observational study: 1) hospitalized acute bipolar patients (n = 205); 2) stable bipolar outpatients (n = 116); 3) hospitalized acute schizophrenia patients (n = 111) and 4) stable schizophrenia outpatients (n = 75), in addition to healthy controls (HC) (n = 185). The quantitative determination of serum kynurenine metabolites was performed using liquid chromatography–tandem mass spectrometry.Kynurenine levels were lower in all patients combined compared to HC while ANCOVA analyses did not reveal inter-diagnostic difference between SCZ and BD. Interestingly, hospitalized patients of both diagnostic groups combined displayed significantly lower kynurenine levels than stabilized outpatients. Psychotic symptoms were associated with lower quinaldic acid (F = 9.18, p=

Irradiating normal-superconducting junctions with microwave photons produce spectacular effects, such as Shapiro steps and photoinduced modifications of the dc supercurrent. Moreover, microwave irradiation can also have other, hitherto unexplored consequences, such as a photoassisted dissipation which is phase dependent. Here we present a finite-frequency measurement of both the dissipation and the supercurrent of a phase-biased graphene-superconductor junction in response to microwave photons. We find that, while the supercurrent response is well described by existing theory, the dissipation exhibits unexpected effects which need new theoretical elucidation. Especially with high frequency photons, the dissipation is enhanced at phase zero, where it is minimum without irradiation. We attribute this enhancement to Andreev level transitions, made possible by microwave-induced nonequilibrium population of Andreev bound states. Our results demonstrate that dissipation is a more sensitive probe of microwave photons than is the supercurrent, and reveal the potential of measuring dissipation to improve superconducting photodetectors and investigate photoassisted physics in hybrid superconducting systems.

Background: Causes of variations in outcomes from cancer care in developed countries are often unclear. Australia has developed health system pathways describing consensus standards of optimal cancer care across the phases of prevention through to follow-up or end-of-life. These Optimal Care Pathways (OCP) were introduced from 2013 to 14. We investigated whether care consistent with the OCP improved outcomes for colon cancer patients. Methods: Colon patients diagnosed from 2008 to 2014 were identified from the Australian State of Victoria Cancer Registry (VCR) and cases linked with State and Federal health datasets. Surrogate variables describe OCP alignment in our cohort, across three phases of the pathway; prevention, diagnosis and initial treatment and end-of-life. We assessed the impact of alignment on (1) stage of disease at diagnosis and (2) overall survival. Findings: Alignment with the prevention phase of the OCP occurred for 88% of 13,539 individuals and was associated with lower disease stage at diagnosis (OR = 0.33, 95% confidence interval 0.24 to 0.42), improved crude three-year survival (69.2% versus 62.2%; p

Abstract Background Diphyllobothriosis is an intestinal cestodosis caused by tapeworms of the family Diphyllobothriidae. In France, endemic cases are limited to south-east and due to Dibothriocephalus latus. In this paper, we investigate a series of seven cases of diphyllobothriosis in the non-endemic French region of Brittany. All have been diagnosed between 2016 and 2018 at the University Hospital of Rennes. Methods Parasites were identified by their morphological features and by phylogenetic analysis of the cox1 gene. Phylogenetic tree was built using maximum likelihood criterion under the GTR+G+I model and 2000 bootstrap replicates. A form was sent to all patients to collect data concerning clinical signs and possible sources of infection. Results All cases were due to Dibothriocephalus nihonkaiensis, a species strictly distributed in the North Pacific. Epidemiological investigation showed that the parasite was probably acquired in France, after consumption of Japanese food containing raw salmon. All patients presented with at least abdominal pain and fatigue except for one patient who had no symptoms. Conclusions To our knowledge, this case series is the most important cohort of allochthonous diphyllobothriosis described in Europe. This sudden emergence raises concern about foodborne infections, highlighting (i) risky food habits in absence of adequate sanitary control; and (ii) the breaking of the rule of geographical restriction due to globalization and worldwide trades.

The diagnosis of intestinal parasitic infections still widely relies on microscopic examination of stools and requires reliable reagents and staff expertise. The ParaFlo® assays (Eurobio Ingen) are ready-to-use concentration methods for parasite egg detection, and they could improve reagent traceability and ease of manipulation. Ninety-three stool samples were analyzed with the ParaFlo® concentration methods and then compared with routine microscopic methods for protozoa and helminth detection: seventy-eight were analyzed with ParaFlo® Bailenger and in-house Thebault or Bailenger concentrations, and fifty-five were analyzed with ParaFlo®DC and the in-house merthiolate-formalin diphasic concentration (DC) method. Fully concordant results were obtained for 75%, 70%, and 69% of samples when comparing ParaFlo® DC and in-house DC, ParaFlo® Bailenger and in-house Bailenger, and ParaFlo® Bailenger and Thebault, respectively. The performances of the ParaFlo® assays did not differ statistically from that obtained with their in-house counterparts (Bailenger and DC) for the detection of protozoa, but ParaFlo® Bailenger performed significantly poorer than the Thebault method (p < 0.001). No statistical differences were observed between the commercial and in-house methods for helminth detection. These marketed concentration methods could be used in routine if combined with other techniques for protozoa detection.

Abstract Background In prospective studies, sugar-sweetened beverages (SSB) have been associated with weight increase. However, most prospective studies examine changes in body weight over time according to amounts of SSB intake at baseline, generally without considering changes over time in SSB, energy intake and physical activity. The objective of the present study was to examine how SSB intakes influence changes in weight, according to the way SSB intakes are analysed. Methods For a prospective cohort study with two nutritional assessments in time, 46 Flemish municipalities were selected by clustered random sampling. Within these municipalities, a random sample of men and women between 18 and 75 years of age was selected and invited to participate. In total, 562 middle-aged Belgian adults were tested in 2002 and 2012 for the same anthropometric, lifestyle, nutrition and physical activity parameters. The main outcome measured were weight changes from 2002 to 2012 were analysed according to various ways to parametrise SSB intakes in 2002 and changes in SSB, energy intake, and physical activity from 2002 to 2012. Results In a multivariable model including age, sex, the best predictors of weight changes were the weight at baseline (P

Abstract Background Observational studies and meta-analyses relating milk consumption by adults to all-cause mortality, coronary heart disease and stroke have obtained contradictory results. Some studies found a protective effect of milk consumption, whilst other found an increased risk. Methods We performed a systematic literature search until June 2015 on prospective studies that looked at milk consumption, all-cause mortality, coronary heart disease and stroke. Random-effect meta-analyses were performed with dose-response. Results Twenty-one studies involving 19 cohorts were included in this meta-analysis, 11 on all-cause mortality, 9 on coronary heart disease, and 10 on stroke. Milk intake ranged from 0 to 850 mL/d. The summary relative risk (SRR) for 200 mL/d milk consumption was 1.01 (95% CI: 0.96–1.06) for all-cause mortality, 1.01 (95% CI: 0.98–1.05) for fatal and non fatal coronary heart disease, and 0.91 (95% CI: 0.82–1.02) for fatal and non fatal stroke. Stratified analyses by age, Body Mass Index, total energy intake and physical acitivity did not alter the SRR estimates. The possibility of publication bias was found for all cause mortality and for stroke, indicating a gap in data that could have suggested a higher risk of these conditions with increased milk consumption. Conclusions We found no evidence for a decreased or increased risk of all-cause mortality, coronary heart disease, and stroke associated with adult milk consumption. However, the possibility cannot be dismissed that risks associated with milk consumption could be underestimated because of publication bias.

Invasive pulmonary aspergillosis (IPA) in intensive care unit patients is a major concern. Influenza-associated acute respiratory distress syndrome (ARDS) and severe COVID-19 patients are both at risk of developing invasive fungal diseases. We used the new international definitions of influenza-associated pulmonary aspergillosis (IAPA) and COVID-19-associated pulmonary aspergillosis (CAPA) to compare the demographic, clinical, biological, and radiological aspects of IAPA and CAPA in a monocentric retrospective study. A total of 120 patients were included, 71 with influenza and 49 with COVID-19-associated ARDS. Among them, 27 fulfilled the newly published criteria of IPA: 17/71 IAPA (23.9%) and 10/49 CAPA (20.4%). Kaplan–Meier curves showed significantly higher 90-day mortality for IPA patients overall (p = 0.032), whereas mortality did not differ between CAPA and IAPA patients. Radiological findings showed differences between IAPA and CAPA, with a higher proportion of features suggestive of IPA during IAPA. Lastly, a wide proportion of IPA patients had low plasma voriconazole concentrations with a higher delay to reach concentrations > 2 mg/L in CAPA vs. IAPA patients (p = 0.045). Severe COVID-19 and influenza patients appeared very similar in terms of prevalence of IPA and outcome. The dramatic consequences on the patients’ prognosis emphasize the need for a better awareness in these particular populations.

Molecular biology has been gaining more importance in parasitology. Recently, a commercial multiplex PCR assay detecting helminths was marketed: the Allplex™ GI-Helminth(I) Assay. It targets Ancylostoma spp., Ascaris spp., Enterobius vermicularis, Hymenolepis spp., Necator americanus, Strongyloides spp., Taenia spp. and Trichuris trichiura, but also the two most common microsporidia genera in human health, i.e. Enterocytozoon spp. and Encephalitozoon spp. This study aimed to evaluate and compare the Allplex™ GI-Helminth(I) Assay to classical diagnostic methods, based on a cohort of 110 stool samples positive for helminths (microscopy) or for microsporidia (PCR). Samples were stored at −80 °C until analysis by the Allplex™ GI-Helminth(I) Assay. False-negatives were re-tested with bead-beating pretreatment. Without mechanical lysis, concordance and agreement between microscopy and Allplex™ GI-Helminth(I) Assay ranged from 91% to 100% and from 0.15 to 1.00, respectively depending on the target. Concordance was perfect for Taenia spp. (n = 5) and microsporidia (n = 10). False-negative results were observed in 54% (6/13), 34% (4/11) and 20% (7/35) of cases, for hookworms, E. vermicularis and Strongyloides spp. detection, respectively. For these targets, pretreatment improved the results, but only slightly. Trichuris trichiura detection was critically low without pretreatment, as only 9% (1/11) of the samples were positive, but detection reached 91% (10/11) with bead-beating pretreatment. Mechanical lysis was also needed for Ascaris spp. and Hymenolepis spp. to reduce false-negative results from 1/8 to 1/21, respectively, to none for both. Overall, with an optimized extraction process, the Allplex™ GI-Helminth(I) Assay allows the detection of numerous parasites with roughly equivalent performance to that of microscopy, except for hookworms.

Strongyloides stercoralis serology is a sensitive method for strongyloidiasis diagnosis, but it is prone to cross-reactions with other helminthiases. This four-year retrospective study aimed at estimating the performance of the Bordier IVD® Strongyloides ratti ELISA assay in a non-endemic country (France). The study included all patients tested for strongyloidiasis in our center between 2015 and 2019, by both serology and stool examination. Cases were defined using an algorithm considering serological results, microscopic examination of stools, and other biological, clinical or epidemiological data. The study included 805 stools from 341 patients (70% migrants, 20% travelers, 10% without travel to a highly endemic area). Thirty patients (8.8%) had positive serology, 9 had microscopically proven strongyloidiasis, and 11 and 10 were classified as probable and possible strongyloidiasis, respectively. Performances of microscopy and serology were compared, considering proven and probable strongyloidiasis as true infections. The sensitivity, specificity, positive predictive value and negative predictive value of serology were 100%, 97%, 67% and 100%, respectively, and those of microscopic examination of stools were 45% (p

This study aims at evaluating the performances of the multiplex PCR AllplexTM Gastrointestinal Panel-Parasite Assay (GIPPA), which detects G. duodenalis, Cryptosporidium spp., E. histolytica, D. fragilis, B. hominis, and C. cayetanensis, by comparison to microscopy. A retrospective evaluation was conducted on a series of positive clinical samples (n = 99) stored at −80 °C or at +4 °C. A five-month prospective study was then conducted on all samples sent to our lab for parasite detection (n = 586). In the retrospective cohort, sensitivity was 81% for both G. duodenalis (26/32) and D. fragilis (21/26) and 100% for Cryptosporidium spp. (26/26, including 6 different species), B. hominis (26/26), and C. cayetanensis (4/4). During the prospective study, 95 samples were positive by microscopy and 207 by multiplex PCR assay. The molecular assay showed a significantly higher sensitivity of PCR, especially for G. duodenalis (100% vs. 60.7%, p < 0.01), D. fragilis (97.2% vs. 14.1%, p < 0.001), and B. hominis (99.4% vs. 44.2%, p < 0.001) but also for E. histolytica (100% vs. 50.0%). The sensitivity of the AllplexTM GIPPA on the first stool sample was equivalent to the sensitivity of microscopy on multiple stool samples but inferior to multiplex PCR on multiple stool samples. Taken together, the AllplexTM GIPPA is suitable for the routine detection of protozoa in fecal samples.

Hyperhomocysteinemia results from hepatic metabolism dysfunction and is characterized by a high plasma homocysteine level, which is also an independent risk factor for cardiovascular disease. Elevated levels of homocysteine in plasma lead to hepatic lesions and abnormal lipid metabolism. Therefore, lowering homocysteine levels might offer therapeutic benefits. Recently, we were able to lower plasma homocysteine levels in mice with moderate hyperhomocysteinemia using an adenoviral construct designed to restrict the expression of DYRK1A, a serine/threonine kinase involved in methionine metabolism (and therefore homocysteine production), to hepatocytes. Here, we aimed to extend our previous findings by analyzing the effect of hepatocyte-specific Dyrk1a gene transfer on intermediate hyperhomocysteinemia and its associated hepatic toxicity and liver dysfunction. Commensurate with decreased plasma homocysteine and alanine aminotransferase levels, targeted hepatic expression of DYRK1A in mice with intermediate hyperhomocysteinemia resulted in elevated plasma paraoxonase-1 and lecithin:cholesterol acyltransferase activities and apolipoprotein A–I levels. It also rescued hepatic apolipoprotein E, J, and D levels. Further, Akt/GSK3/cyclin D1 signaling pathways in the liver of treated mice were altered, which may help prevent homocysteine-induced cell cycle dysfunction. DYRK1A gene therapy could be useful in the treatment of hyperhomocysteinemia in populations, such as end-stage renal disease patients, who are unresponsive to B-complex vitamin therapy.

Summary: Background: Explosive remnants of war (ERW)—landmines, unexploded ordnance (UXO), and abandoned explosive ordnance (AXO)—have been recognised as a threat to health since the 1990s. We aimed to study the effect of ERW on global public health. Methods: In this systematic mixed-studies review, we searched the Web of Science, Scopus, PubMed, and ProQuest databases, and hand searched relevant websites, for articles published between Jan 1, 1990, and Aug 31, 2015. We used keywords and Medical Subject Headings related to ERW, landmines, UXO, and AXO to locate original peer-reviewed quantitative, qualitative, or mixed-methods studies in English of the direct physical or psychological effects of ERW on direct victims of the explosive device or reverberating social and economic effects on direct victims and indirect victims (their families and the wider at-risk community). We excluded studies if more than 20% of participants were military, if they were of deminers, if they were from high-income countries, or if they were of chemical weapons. We identified no peer-reviewed studies of AXO effects, so we extended the search to include grey literature. We critically appraised study quality using a mixed methods appraisal tool. We used a narrative synthesis approach to categorise and synthesise the literature. We extracted quantitative data and calculated means and percentages. Findings: The initial search identified 10 226 studies, leaving 8378 (82%) after removal of duplicates, of which we reviewed 54 (26 [48%] were quantitative descriptive studies, 20 [37%] were quantitative non-randomised studies, four [7%] were mixed-methods studies, and four [7%] were grey literature). The direct psychological effects of landmines or UXO appear high. We identified comorbidity of anxiety and depression in landmine or UXO victims in four studies, more women presented with post-traumatic stress disorder than did men in two studies, and landmine or UXO victims reported a greater prevalence of post-traumatic stress disorder, anxiety, or depression than did control groups in two studies. Overall injury and mortality rates caused by landmines or UXO decreased over time across five studies and increased in one. More men were injured or killed by landmines or UXO than were women (0–30·6% of women), the mean ages of casualties ranged from 18·5 years to 38·1 years, and victims were likely to be doing an activity of economic necessity at the time of injury. The proportion of casualties of landmines or UXO younger than 18 years ranged from 22% to 55% across twelve studies. Landmine or UXO victims who had one or more limbs amputated ranged from 19·5% to 82·6%. Landmines and UXO had a negative effect on internally displaced populations and returning refugees, physical security, economic productivity, child health and educational attainment, food security, and agriculture in studies from seven countries. We could not establish the proportion of casualties caused by AXO from unplanned explosions at munitions sites, although the grey literature suggests that AXO is a substantial problem. Interpretation: Individually, these landmine and UXO results are not new and substantiate findings from existing research. Taken together, however, these findings provide a picture of the effect of landmines and UXO that stretches far beyond injury and mortality prevalence, making landmine and UXO clearance a more favourable option for funders. AXO effects are understudied and warrant further research. Funding: King's College London.

BACKGROUND:The role of breast screening in breast cancer mortality declines is debated. Screening impacts cancer mortality through decreasing the number of advanced cancers with poor diagnosis, while cancer treatment works through decreasing the case-fatality rate. Hence, reductions in cancer death rates thanks to screening should directly reflect reductions in advanced cancer rates. We verified whether in breast screening trials, the observed reductions in the risk of breast cancer death could be predicted from reductions of advanced breast cancer rates. PATIENTS AND METHODS:The Greater New York Health Insurance Plan trial (HIP) is the only breast screening trial that reported stage-specific cancer fatality for the screening and for the control group separately. The Swedish Two-County trial (TCT)) reported size-specific fatalities for cancer patients in both screening and control groups. We computed predicted numbers of breast cancer deaths, from which we calculated predicted relative risks (RR) and (95% confidence intervals). The Age trial in England performed its own calculations of predicted relative risk. RESULTS:The observed and predicted RR of breast cancer death were 0.72 (0.56-0.94) and 0.98 (0.77-1.24) in the HIP trial, and 0.79 (0.78-1.01) and 0.90 (0.80-1.01) in the Age trial. In the TCT, the observed RR was 0.73 (0.62-0.87), while the predicted RR was 0.89 (0.75-1.05) if overdiagnosis was assumed to be negligible and 0.83 (0.70-0.97) if extra cancers were excluded. CONCLUSIONS:In breast screening trials, factors other than screening have contributed to reductions in the risk of breast cancer death most probably by reducing the fatality of advanced cancers in screening groups. These factors were the better management of breast cancer patients and the underreporting of breast cancer as the underlying cause of death. Breast screening trials should publish stage-specific fatalities observed in each group.

Although microscopic examination of stool samples remains the reference method for the diagnosis of intestinal protozoal infections, these techniques are time-consuming and require operators who are experienced and well trained. Molecular biology seems to offer performances at least equivalent in terms of sensitivity and specificity for certain parasites. This study aimed to compare three multiplex PCR assays on 93 prospectively collected positive stools (prospective cohort) and a panel of 12 more Cryptosporidium-positive samples (Cryptosporidium panel). On the prospective cohort, the sensitivity was 89%, 64% and 41% for Giardia sp. detection for BD MaxTM, G-DiaParaTM and RIDA®GENE, respectively and 75%, 100% and 100% for C. parvum/hominis detection. The sensitivity of the RIDA®GENE assay for all Cryptosporidium species was 100%, and for D. fragilis 71%. All the techniques obtained the same results for E. histolytica detection, with one positive sample. All species in the Cryptosporidium panel were identified by the RIDA®GENE PCR. The BD MaxTM and G-DiaParaTM assays detected only C. parvum/hominis with the exception of one positive sample for C. meleagridis. No assay showed satisfactory results for all parasites simultaneously, and the DNA extraction seems to be the critical step. More studies are needed to standardize this procedure.