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OBJECTIVES: The quality control of serological assays remains controversial. The aim of this project was to describe the problems associated with a working model for controlling these assays and solutions, including using a source of well-defined targets and acceptable limits, a process to identify lot-to-lot reagent variation and an interpretation of the result that accounted for the clinical situation. False-negative results are problematic but can be reduced by identifying and comparing reagent lot variation with previous results. METHODS: The components of the Quality Assurance strategy are the following: Lot-to-lot reagent and calibrator variation assessment; dynamic, big-data approach to determine accurate targets and acceptable limits for manufacturer-provided QC material; negative QC monitoring process; use of commutable EQA with a sufficient method subgroup size to assess bias; clinical assessment of any statistically flagged error; and provision of support to the clinician for the interpretation of results. RESULTS: The model described has been used for twelve months, and acceptable variation has been maintained. CONCLUSIONS: The paper presents a solution that emphasizes the early detection of reagent lot variation and patient risk rather than instrument control. Reducing the risk of a false result to patients requires optimal assay quality control and an effective mechanism to support the clinician's use of these results in diagnosis and monitoring. The problems of serological assays are well-known, but there remain few integrated solutions in the literature.
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Aphanomyces euteiches is the most damaging soilborne pea pathogen in France. Breeding of pea resistant varieties combining a diversity of quantitative trait loci (QTL) is a promising strategy considering previous research achievements in dissecting polygenic resistance to A. euteiches. The objective of this study was to provide an overview of the diversity of QTL and marker haplotypes for resistance to A. euteiches, by integrating a novel QTL mapping study in advanced backcross (AB) populations with previous QTL analyses and genome-wide association study (GWAS) using common markers. QTL analysis was performed in two AB populations derived from the cross between the susceptible spring pea variety "Eden" and the two new sources of partial resistance "E11" and "LISA". The two AB populations were genotyped using 993 and 478 single nucleotide polymorphism (SNP) markers, respectively, and phenotyped for resistance to A. euteiches in controlled conditions and in infested fields at two locations. GWAS and QTL mapping previously reported in the pea-Aphanomyces collection and from four recombinant inbred line (RIL) populations, respectively, were updated using a total of 1,850 additional markers, including the markers used in the Eden x E11 and Eden x LISA populations analysis. A total of 29 resistance-associated SNPs and 171 resistance QTL were identified by GWAS and RIL or AB QTL analyses, respectively, which highlighted 10 consistent genetic regions confirming the previously reported QTL. No new consistent resistance QTL was detected from both Eden x E11 and Eden x LISA AB populations. However, a high diversity of resistance haplotypes was identified at 11 linkage disequilibrium (LD) blocks underlying consistent genetic regions, especially in 14 new sources of resistance from the pea-Aphanomyces collection. An accumulation of favorable haplotypes at these 11 blocks was confirmed in the most resistant pea lines of the collection. This study provides new SNP markers and rare haplotypes associated with the diversity of Aphanomyces root rot resistance QTL investigated, which will be useful for QTL pyramiding strategies to increase resistance levels in future pea varieties.
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ELISA methods are the diagnostic tools recommended for the serological diagnosis of Coxiella burnetii infection in ruminants but their respective diagnostic performances are difficult to assess because of the absence of a gold standard. This study focused on three commercial ELISA tests with the following objectives (1) assess their sensitivity and specificity in sheep, goats and cattle, (2) assess the between- and within-herd seroprevalence distribution in these species, accounting for diagnostic errors, and (3) estimate optimal sample sizes considering sensitivity and specificity at herd level. We comparatively tested 1413 cattle, 1474 goat and 1432 sheep serum samples collected in France. We analyzed the cross-classified test results with a hierarchical zero-inflated beta-binomial latent class model considering each herd as a population and conditional dependence as a fixed effect. Potential biases and coverage probabilities of the model were assessed by simulation. Conditional dependence for truly seropositive animals was high in all species for two of the three ELISA methods. Specificity estimates were high, ranging from 94.8% [92.1; 97.8] to 99.2% [98.5; 99.7], whereas sensitivity estimates were generally low, ranging from 39.3 [30.7; 47.0] to 90.5% [83.3; 93.8]. Between- and within-herd seroprevalence estimates varied greatly among geographic areas and herds. Overall, goats showed higher within-herd seroprevalence levels than sheep and cattle. The optimal sample size maximizing both herd sensitivity and herd specificity varied from 3 to at least 20 animals depending on the test and ruminant species. This study provides better interpretation of three widely used commercial ELISA tests and will make it possible to optimize their implementation in future studies. The methodology developed may likewise be applied to other human or animal diseases.
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Doenças dos Bovinos/diagnóstico , Coxiella burnetii/isolamento & purificação , Ensaio de Imunoadsorção Enzimática/veterinária , Doenças das Cabras/diagnóstico , Febre Q/veterinária , Doenças dos Ovinos/diagnóstico , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/microbiologia , Feminino , França/epidemiologia , Doenças das Cabras/epidemiologia , Doenças das Cabras/microbiologia , Cabras , Análise de Classes Latentes , Prevalência , Febre Q/diagnóstico , Febre Q/epidemiologia , Febre Q/microbiologia , Estudos Soroepidemiológicos , Ovinos , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/microbiologia , Carneiro DomésticoRESUMO
BACKGROUND: Among the strategies to encourage pregnant women to be vaccinated against pertussis in the postpartum period, that of giving them a prescription has been evaluated only sparsely. OBJECTIVE: To measure the effect of giving women who are not immunized against pertussis a prescription for the vaccine at discharge from the maternity unit. MATERIAL AND METHODS: Single-center before-and-after study (2011: before; 2015: after). All women received both oral and written information about vaccination against pertussis. During the after period, they were also specifically asked their immunization status during pregnancy. Those currently unimmunized received a written prescription for it at discharge. RESULTS: Among the women unimmunized at delivery, the percentage who were vaccinated postpartum climbed from 17 to 42% between 2011 and 2015 (p < 0.001), while the percentage of their unimmunized partners who were vaccinated remained stable (27 and 29%, p = 0.74). During this time, the percentage of women immunized against pertussis at the beginning of pregnancy rose from 32 to 52% (p < 0.001). Finally, the percentage of all women protected against this disease postpartum climbed from 44 to 72% between these two periods (p < 0.001). CONCLUSIONS: In the postpartum period, giving a prescription for pertussis vaccine to women unimmunized is accompanied by a significant elevation in their vaccination rate. Nevertheless, this rate remains low and better strategies have to be implemented.
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Estudos Controlados Antes e Depois , Vacina contra Coqueluche/administração & dosagem , Período Pós-Parto , Prescrições , Vacinação/estatística & dados numéricos , Coqueluche/prevenção & controle , Adulto , Saúde da Família , Feminino , Conhecimentos, Atitudes e Prática em Saúde , Humanos , Paridade , Alta do Paciente , Gravidez , Cônjuges/estatística & dados numéricos , Inquéritos e QuestionáriosRESUMO
BACKGROUND: Although its role in the prognosis for delivery remains controversial, the type of breech is sometimes taken into account in the decision about mode of delivery. Objective of our study was to compare maternal and neonatal morbidity for trial of vaginal delivery according to the type of breech (complete or frank). MATERIAL AND METHOD: Single-center retrospective study of women with trials of vaginal delivery of a singleton fetus in breech presentation at of after 37 weeks of gestation. Neonatal status was assessed by the composite variable of the Term Breech Trial, first considered alone, and then with the addition of a 5-min Apgar score < 7 or a neonatal arterial pH<7.0. RESULTS: Of the 495 trials of vaginal delivery during the study period, approximately one third of them were complete breech (35.8 %) and two thirds frank (64.2 %). The frequency of cesareans during labor was similar regardless of the type of breech (16.4 for complete vs 12.6 % for frank, p=0.24), nor did neonatal morbidity differ (1.7 for complete vs 4.1 % for frank, p=0.15). On the other hand, cord prolapse occurred almost exclusively in complete breech presentations (4.5 vs 0.3 %, p<0.01), and prognosis was good in all cases. Complete breech presentations were also associated with more frequent use of forceps to the after-coming head (16.2 vs 9.7 %, p<0.05). Finally, there were non significant difference between the two types of breech concerning severe acidosis but it seemed to have it more in frank breech (2.3 vs 1.2 %, p=0.34). CONCLUSION: Among women eligible for vaginal delivery, the type of breech presentation (complete or frank) has little influence on delivery maternal and neonatal morbidity. The type of breech need not be taken into account in deciding the route of delivery.
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Apresentação Pélvica/classificação , Apresentação Pélvica/epidemiologia , Parto Obstétrico/métodos , Resultado da Gravidez , Adulto , Índice de Massa Corporal , Apresentação Pélvica/terapia , Cesárea/estatística & dados numéricos , Feminino , Idade Gestacional , Humanos , Recém-Nascido , Gravidez , Prognóstico , Estudos RetrospectivosRESUMO
BACKGROUND: Although vaccination of pregnant women against influenza is recommended, the vaccination rate remains low. We conducted a study to identify determinants of influenza vaccination uptake in pregnancy in order to identify strategies to improve seasonal influenza vaccination rates. METHODS: Prospective observational hospital-based study in the French hospital performing the highest number of deliveries, located in the city of Lille, among all women who had given birth during the 2014-2015 influenza season. Data were collected through a self-completed questionnaire and from medical files. The vaccination uptake was self-reported. Determinants of vaccination uptake were identified using logistic regression analysis. RESULTS: Of the 2045 women included in the study, 35.5% reported that they had been vaccinated against influenza during their pregnancy. The principal factors significantly associated with greater vaccination uptake were previous influenza vaccination (50.9% vs 20.2%, OR 4.1, 95% CI 3.1-5.5), nulliparity (41.0% vs 31.3%, OR 2.5, 95% CI 1.7-3.7), history of preterm delivery < 34 weeks (43.4% vs 30.3%, OR 2.3, 95% CI 1.1-4.9), the mother's perception that the frequency of vaccine complications for babies is very low (54.6% vs 20.6%, OR 1.1, 95% CI 0.5-2.2), the mother's good knowledge of influenza and its vaccine (61.7% vs 24.4%, OR 3.1, 95% CI 2.2-4.4), hospital-based prenatal care in their first trimester of pregnancy (55.0% vs 30.2%, OR 2.1, 95% CI 1.2-3.7), vaccination recommendations during pregnancy by a healthcare worker (47.0% vs 2.7%, OR 18.8, 95% CI 10.0-35.8), receipt of a vaccine reimbursement form (52.4% vs 18.6%, OR 2.0, 95% CI 1.5-2.7), and information from at least one healthcare worker about the vaccine (43.8% vs 19.1%, OR 1.8, 95% CI 1.3-2.6). CONCLUSIONS: Our findings suggest that in order to increase flu vaccination compliance among pregnant women, future public health programmes must ensure cost-free access to vaccination, and incorporate education about the risks of influenza and the efficacy/safety of vaccination and clear recommendations from healthcare professionals into routine antenatal care.
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Conhecimentos, Atitudes e Prática em Saúde , Vacinas contra Influenza/uso terapêutico , Influenza Humana/prevenção & controle , Cooperação do Paciente , Gestantes , Adulto , Feminino , França , Gastos em Saúde , Humanos , Modelos Logísticos , Paridade , Educação de Pacientes como Assunto , Gravidez , Cuidado Pré-Natal , Estudos Prospectivos , Adulto JovemRESUMO
Coxiella burnetii can infect many animal species, but its circulation dynamics in and through horses is still unclear. This study evaluated horse exposure in an area known to be endemic for ruminants and humans. We assessed antibody prevalence in horse serum by ELISA, and screened by qPCR horse blood, ticks found on horses and dust from stables. Horse seroprevalence was 4% (nâ¯=â¯335, 37 stables) in 2015 and 12% (nâ¯=â¯294, 39 stables) in 2016. Of 199 horses sampled both years, 13 seroconverted, eight remained seropositive, and one seroreverted. Seropositive horses were located close to reported human cases, yet none displayed Q fever-compatible syndromes. Coxiella DNA was detected in almost 40% of collected ticks (nâ¯=â¯59/148 in 2015; nâ¯=â¯103/305 in 2016), occasionally in dust (nâ¯=â¯3/46 in 2015; nâ¯=â¯1/14 in 2016) but never in horse blood. Further studies should be implemented to evaluate if horses may be relevant indicators of zoonotic risk in urban and suburban endemic areas.
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Anticorpos Antibacterianos/sangue , Coxiella burnetii/fisiologia , Doenças dos Cavalos/epidemiologia , Febre Q/veterinária , Animais , Coxiella burnetii/genética , DNA Bacteriano/genética , Ensaio de Imunoadsorção Enzimática , Doenças dos Cavalos/sangue , Cavalos , Reação em Cadeia da Polimerase , Febre Q/sangue , Febre Q/epidemiologia , Estudos Soroepidemiológicos , Carrapatos/microbiologiaRESUMO
RASA1-related disease is a rare autosomal dominant disease characterized by capillary malformations, arteriovenous malformations (AVMs), and/or arteriovenous fistulas (AFVs). Penetrance is nearly complete and vascular malformations may cause serious complications such as organ injury due to oxygenation disorder, brain abscess, hemorrhage, and stroke. Early diagnosis is useful in order to discuss optimal management, including AVMs/AVFs embolization or surgical procedures, and try to prevent some of the complications. In this context, molecular testing of RASA1 gene mutation in relatives may help to better manage the family. All arteriovenous malformations are however not accessible to such procedures. In addition, these therapeutic procedures may result in potential side effects and complications. A couple was referred to our genetics unit and asked us for prenatal genetic testing about a RASA1 mutation. Here, we discuss about arguments that led our team to accept prenatal testing. To the best of our knowledge, no molecular prenatal diagnosis was reported until now in RASA1-related diseases. This first report of prenatal diagnosis in RASA1-related diseases may also offer perspectives for a more general discussion in the field of inherited arteriovenous malformations.
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Testes Genéticos , Diagnóstico Pré-Natal , Proteína p120 Ativadora de GTPase/genética , Adulto , Feminino , Humanos , Masculino , GravidezRESUMO
Mass spectrometry (MS) in Selected Reaction Monitoring (SRM) mode is proposed for in-depth characterisation of microorganisms in a multiplexed analysis. Within 60-80 minutes, the SRM method performs microbial identification (I), antibiotic-resistance detection (R), virulence assessment (V) and it provides epidemiological typing information (T). This SRM application is illustrated by the analysis of the human pathogen Staphylococcus aureus, demonstrating its promise for rapid characterisation of bacteria from positive blood cultures of sepsis patients.
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Bactérias/classificação , Bactérias/efeitos dos fármacos , Técnicas de Tipagem Bacteriana , Espectrometria de Massas/métodos , Bactérias/patogenicidade , Técnicas de Tipagem Bacteriana/métodos , Farmacorresistência Bacteriana , Humanos , Reprodutibilidade dos Testes , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/classificação , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/patogenicidade , Virulência/genéticaRESUMO
In recent decades, concern has been growing about decreasing fecundity and fertility in the human population. Exposure to non-ionizing electromagnetic fields (EMF), especially radiofrequency (RF) fields used in wireless communications has been suggested as a potential risk factor. For the first time, we evaluated the effects of exposure to the 2450MHz Wi-Fi signal (1h/day, 6days/week) on the reproductive system of male and female Wistar rats, pre-exposed to Wi-Fi during sexual maturation. Exposure lasted 3 weeks (males) or 2 weeks (females), then animals were mated and couples exposed for 3 more weeks. On the day before delivery, the fetuses were observed for lethality, abnormalities, and clinical signs. In our experiment, no deleterious effects of Wi-Fi exposure on rat male and female reproductive organs and fertility were observed for 1h per days. No macroscopic abnormalities in fetuses were noted, even at the critical level of 4W/kg.
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Desenvolvimento Embrionário/efeitos da radiação , Desenvolvimento Fetal/efeitos da radiação , Infertilidade Feminina/etiologia , Infertilidade Masculina/etiologia , Ondas de Rádio/efeitos adversos , Maturidade Sexual/efeitos da radiação , Tecnologia sem Fio , Animais , Relação Dose-Resposta à Radiação , Implantação do Embrião/efeitos da radiação , Perda do Embrião/etiologia , Ingestão de Energia/efeitos da radiação , Feminino , Genitália Masculina/crescimento & desenvolvimento , Genitália Masculina/imunologia , Genitália Masculina/efeitos da radiação , Masculino , Exposição Materna/efeitos adversos , Tamanho do Órgão/efeitos da radiação , Ovário/crescimento & desenvolvimento , Ovário/imunologia , Ovário/efeitos da radiação , Exposição Paterna/efeitos adversos , Distribuição Aleatória , Ratos , Ratos WistarRESUMO
We report the synthesis and characterization of tantalum-boronate trimetallic clusters of general formula {[Cp*Ta](3)(µ(2)-RB(O)(2))(3)(µ(2)-OH)(µ(2)-O)(2)(µ(3)-OH)} (R= 4-(C(6)H(5))(C(6)H(4)) (Ta(3)-4Ph), 4-(C(6)H(5)O)(C(6)H(4)) (Ta(3)-4OPh), 4-(C(7)H(7)O)(C(6)H(4)) (Ta(3)-4OBn), 4-(C(8)H(5))(C(6)H(4)) (Ta(3)-4PhEt), and 4-(C(12)H(7))(C(6)H(4)) (Ta(3)-4Napht)). All complexes have been characterized by NMR spectroscopy and X-ray diffraction. The trimetallic species feature a large Lewis acid type cavity allowing for substrate binding in both the solid and the liquid state using a unique electrostatic interaction and a hydrogen bond. ΔH° and ΔS° values for association of acetone with the complexes vary between -2.0 and -4.1 kcal·mol(-1) and -3 and 2 cal·mol(-1)·K(-1), respectively, showing weaker binding than smaller cavitands of the same type. The barrier for acetone exchange at equilibrium is similar for all complexes, and ΔH() values vary between 8.2 and 11.4 kcal·mol(-1).
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The steroid nucleus is an interesting scaffold for the development of new therapeutic agents. Within the goal of identifying anticancer agents, new pregnane derivatives were prepared by using a sequence of liquid and solid-phase reactions. After we dehydrated epi-allopregnanolone in one step with diethylaminosulfur trifluoride and generated a 2,3α-epoxide, the regio- and stereo-selective aminolysis of this epoxide enabled us to obtain a 2ß-piperazino-pregnane, whose secondary amine was protected as N-Fmoc-derivative. Using the difference in reactivity between OHs 3 and 20, we linked the pregnane nucleus-selectively on the polystyrene diethylbutylsilane resin via the OH in position 20. We next achieved in parallel the coupling of an amino acid (1st level of diversity) and the coupling of a carboxylic acid (2nd level of diversity) to generate two libraries of pregnane derivatives. The compounds inhibited the HL-60 leukemia cell growth and the most potent were three compounds (PD, LPC-37 and LPC-48) with a l-proline as first level of diversity and a cyclohexyl-carbonyl, a naphthalene-2-carbonyl or a 3-acetylbenzoyl as second level of diversity. LPC-48 efficiently inhibited HL-60 cell proliferation with IC(50) value of 1.9 µM and exhibited a low toxicity on normal peripheral blood lymphocytes (IC(50)=31 µM). These results encouraged us to further evaluate the biological activity of these new aminosteroids by investigating their preliminary mechanism of action.
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Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Leucemia/patologia , Pregnanos/síntese química , Pregnanos/farmacologia , Antineoplásicos/química , Antineoplásicos/uso terapêutico , Morte Celular/efeitos dos fármacos , Núcleo Celular/efeitos dos fármacos , Fragmentação do DNA/efeitos dos fármacos , Dietilaminas/química , Flúor/química , Pontos de Checagem da Fase G1 do Ciclo Celular/efeitos dos fármacos , Células HL-60 , Humanos , Concentração Inibidora 50 , Leucemia/tratamento farmacológico , Pregnanos/química , Pregnanos/uso terapêutico , Fase de Repouso do Ciclo Celular/efeitos dos fármacos , Bibliotecas de Moléculas Pequenas/síntese química , Bibliotecas de Moléculas Pequenas/química , Bibliotecas de Moléculas Pequenas/farmacologia , Bibliotecas de Moléculas Pequenas/uso terapêutico , Técnicas de Síntese em Fase SólidaRESUMO
BACKGROUND: The increase in exposure to the Wireless Fidelity (Wi-Fi) wireless communication signal has raised public health concerns especially for young people. Animal studies looking at the effects of early life and prenatal exposure to this source of electromagnetic fields, in the radiofrequency (RF) range, on development and behavior have been considered as high priority research needs by the World Health Organization. METHODS: For the first time, our study assessed the effects of in utero exposure to a 2450 MHz Wi-Fi signal (2 hr/day, 6 days/week for 18 days) on pregnant rats and their pups. Three levels in terms of whole-body specific absorption rate were used: 0.08, 0.4, and 4 W/kg. The prenatal study on fetuses delivered by caesarean (P20) concerned five females/group. The dams and their offspring were observed for 28 days after delivery (15 females/group). RESULTS: For all test conditions, no abnormalities were noted in the pregnant rats and no significant signs of toxicity were observed in the pre- and postnatal development of the pups, even at the highest level of 4 W/kg. CONCLUSIONS: In the present study, no teratogenic effect of repeated exposures to the Wi-Fi wireless communication signal was demonstrated even at the highest level of 4 W/kg. The results from this screening study aimed at investigating Wi-Fi effects, strengthen the previous conclusions that teratology and development studies have not detected any noxious effects of exposures to mobile telephony-related RF fields at exposure levels below standard limits.
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Campos Eletromagnéticos/efeitos adversos , Efeitos Tardios da Exposição Pré-Natal/patologia , Monitoramento de Radiação/métodos , Ondas de Rádio/efeitos adversos , Animais , Animais Recém-Nascidos/crescimento & desenvolvimento , Feminino , Gravidez , Ratos , Ratos Wistar , Reprodução , Testes de Toxicidade , Tecnologia sem FioRESUMO
UNLABELLED: Uterine artery Doppler anomalies are associated with unfavorable outcomes in women with preeclampsia. OBJECTIVES: To examine the association between abnormal uterine artery Doppler and severity of preeclampsia. STUDY DESIGN: Retrospective analysis of a population of 287 patients with preeclampsia who underwent a uterine artery Doppler velocimetric examination at the onset of preeclampsia, between 1996 and 2002. The population was classified into three groups: Group I had normal uterine artery Doppler findings; Group II had a high uterine artery resistance index; and, Group III had both high uterine artery resistance index and bilateral notches. RESULTS: Compared to Group I, preeclampsia occurred earlier in Group II (76.3%) (p < 0.001), and HELLP syndrome was most frequent in Group III. The frequency of fetal growth restriction (pound 3(rd) percentile) was different between groups (19.1, 32.4 and 49.7% in groups I, II and III, respectively). The perinatal death rate was significantly higher in group III than the other two groups (12.8% vs 2.9%, p < 0.01). CONCLUSIONS: In patients with preeclampsia, the presence of uterine artery Doppler anomalies (high resistance index with or without bilateral notches) was associated with unfavorable pregnancy outcomes.
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Pré-Eclâmpsia/diagnóstico por imagem , Útero/irrigação sanguínea , Adulto , Artérias/diagnóstico por imagem , Artérias/fisiopatologia , Feminino , Humanos , Fluxometria por Laser-Doppler , Pré-Eclâmpsia/fisiopatologia , Gravidez , Resultado da Gravidez , Estudos Retrospectivos , Ultrassonografia Doppler em Cores , Útero/diagnóstico por imagem , Adulto JovemRESUMO
Q fever is a zoonosis caused by Coxiella burnetii, a bacterium largely carried by ruminants and shed into milk, vaginal mucus, and feces. The main potential hazard to humans and animals is due to shedding of bacteria that can then persist in the environment and be aerosolized. The purpose of this study was to evaluate shedding after an outbreak of Q fever abortion in goat herds and to assess the relationship with the occurrence of abortions and antibody responses. Aborting and nonaborting goats were monitored by PCR for C. burnetii shedding 15 and 30 days after the abortion episodes. PCR analysis of all samples showed that 70% (n = 50) of the aborting and 53% (n = 70) of the nonaborting goats were positive. C. burnetii was shed into vaginal mucus, feces, and milk of 44%, 21%, and 38%, respectively, of goats that aborted and 27%, 20%, and 31%, respectively, of goats that delivered normally. Statistical comparison of these shedding results did not reveal any difference between these two groups. PCR results obtained for the vaginal and fecal routes were concordant in 81% of cases, whereas those for milk correlated with only 49% of cases with either vaginal or fecal shedding status. Serological analysis, using enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA), and complement fixation tests, showed that at least 24% of the seronegative goats shed bacteria. Positive vaginal and fecal shedding, unlike positive milk shedding, was observed more often in animals that were weakly positive or negative by ELISA or IFA. Two opposite shedding trends were thus apparent for the milk and vaginal-fecal routes. Moreover, this study showed that a nonnegligible proportion of seronegative animals that delivered normally could excrete C. burnetii.
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Coxiella burnetii/imunologia , Coxiella burnetii/isolamento & purificação , Surtos de Doenças , Doenças das Cabras/epidemiologia , Doenças das Cabras/microbiologia , Febre Q/veterinária , Animais , Anticorpos Antibacterianos/sangue , Testes de Fixação de Complemento , Ensaio de Imunoadsorção Enzimática , Fezes/microbiologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Cabras , Leite/microbiologia , Reação em Cadeia da Polimerase/métodos , Febre Q/epidemiologia , Febre Q/microbiologia , Vagina/microbiologiaRESUMO
Coxiella burnetii is an intracellular bacterium that causes a worldwide zoonosis, the Q fever. Currently, to diagnose the infection in ruminants, whole cell antigens-based ELISAs are used. In this study a heat shock protein, the HspB, was evaluated for its ability to be recognized by the goat immune system and its capacity to sign a stage of infection. The htpB gene of C. burnetii was cloned and sequenced. A high identity (>90%) was observed among the htpB genes of four ruminant strains tested. A recombinant protein was expressed in a prokaryotic expression system. The rHspB protein was used to determine the IgG reactivity by ELISA. Sera from experimentally and naturally infected goats were tested. The rHspB is recognized early during the infection course, at 18 days post-infection. Moreover, 80-90% of the animals tested were positive at 39-60dpi. In addition, animals presenting a reactivation of the infection displayed a higher reactivity, statistically significant (p<0.05), than that of the animals in latent infection. These findings suggest that the rHspB could be a good candidate for the development of an ELISA test making possible the detection of recent C. burnetii infection in goats as well as reactivation in those with latent infection.
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Antígenos de Bactérias/imunologia , Doenças das Cabras/diagnóstico , Proteínas de Choque Térmico/imunologia , Testes Imunológicos/veterinária , Febre Q/diagnóstico , Animais , Anticorpos Antibacterianos/sangue , Proteínas de Bactérias/imunologia , Doenças das Cabras/sangue , Doenças das Cabras/microbiologia , Cabras , Testes Imunológicos/métodos , Febre Q/sangue , Febre Q/microbiologia , Fatores de TempoRESUMO
Eutrophication, and the resulting proliferation of phytoplankton can affect water quality. Although environmental and ecological processes that lead to phytoplankton proliferations in temperate areas have received considerable attention, scientific background regarding this topic in tropical areas, especially West Africa, are scarce. In this study, bioassays in batch cultures were carried out in order to identify factors that may be stimulating or limiting the biomass of phytoplankton at various sites in the lower Senegal River delta region. Complete factorial designs were used to test several factors both alone and combined (nitrogen, phosphorus, sulfate, EDTA and trace elements, stirring). The effects of these factors on the growth of the whole phytoplankton communities were followed by the daily measurement of in vivo fluorescence. At all but two of the sites, enrichment with certain nutrients significantly stimulated phytoplankton biomass growth, with nitrogen and/or phosphorus appearing to be the primary limiting factor(s). Nitrogen was the primary limiting factor in coastal ocean waters and in the Senegal River estuary. In the freshwater ecosystems studied, nitrogen and phosphorus acted as alternating or concomitant limiting factors. We assessed the changes in the phytoplankton composition induced by nutrient enrichments in Lake Guiers, a drinking water reservoir for the population of Dakar. Our findings lead us to define the conditions of a possible proliferation of the potentially toxic cyanobacterium Cylindrospermopsis raciborskii mainly in response to phosphorus inputs.
Assuntos
Cianobactérias/crescimento & desenvolvimento , Eutrofização , Nitrogênio/farmacologia , Fósforo/farmacologia , Fitoplâncton/crescimento & desenvolvimento , Biomassa , Quelantes/farmacologia , Cianobactérias/efeitos dos fármacos , Ácido Edético/farmacologia , Água Doce , Metais/farmacologia , Fitoplâncton/efeitos dos fármacos , Água do Mar , SenegalRESUMO
Germanicane-type triterpenes allobetulin (3) and 28-oxoallobetulin (4) can be obtained by the Wagner-Meerwein rearrangement of the more available lupane-type triterpenes betulin (1) and betulinic acid (2), respectively. The medical uses of betulinic acid (2) and its derivatives are limited because of their poor hydrosolubility and pharmacokinetics properties. In order to overcome this major problem, we synthesized and studied the in vitro anticancer activity of a series of 3beta-O-monodesmosidic saponins derived from betulin (14-16), betulinic acid (20-22), allobetulin (23-28) and 28-oxoallobetulin (29-34) based on six different natural sugar residues (d-glucose, l-rhamnose, d-arabinose, d-galactose, d-mannose and d-xylose). This structure-activity relationship study confirmed that betulinic acid saponins are generally better in vitro anticancer agents than those derived from betulin with the exception of betulin 3beta-O-alpha-d-mannopyranoside (15) which exerted a potent cytotoxic activity against lung carcinoma (A-549) and colorectal adenocarcinoma (DLD-1) human cell lines with IC(50) ranging from 7.3 to 10.1mumol/L. Furthermore, although the synthesis of novel germanicane-type saponins was carried out with success, the bioactivity measured for these glycosides was not as high as we anticipated since only the 3beta-O-beta-d-glucopyranoside and 3beta-O-beta-d-galactopyranoside of allobetulin (23,24) showed moderate anticancer activity (IC(50) 30-40 micromol/L).
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Pulmonares/tratamento farmacológico , Saponinas/síntese química , Triterpenos/química , Adenocarcinoma/tratamento farmacológico , Adenocarcinoma/patologia , Antineoplásicos Fitogênicos/síntese química , Antineoplásicos Fitogênicos/química , Betula/química , Células Cultivadas/efeitos dos fármacos , Neoplasias Colorretais/patologia , Ensaios de Seleção de Medicamentos Antitumorais , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Humanos , Neoplasias Pulmonares/patologia , Estrutura Molecular , Saponinas/química , Saponinas/farmacologia , Relação Estrutura-AtividadeRESUMO
Performances of an ELISA, an immunofluorescence assay (IFA) and a complement fixation test (CFT) were assessed for detecting antibodies against Coxiella burnetii after Q fever abortions in naturally infected goats. The goal of the study was to provide information useful for veterinary serodiagnosis in regard to categories of goats either experiencing Q fever abortion or not, blood sampling times and recommended cut-offs. The study was conducted on eight goat herds with evidence of C. burnetii abortions. In each herd, at least 5 goats that had aborted and 10 goats prior to parturition or at term were monitored 15, 30 and 60 days (D15, D30, D60) after the onset of Q fever abortion. The overall CFT results distribution did not differ between the two groups of goats and showed poor agreement with the ELISA results. In contrast, the ELISA and IFA results revealed comparable significant differences, but overall the ELISA test was slightly more sensitive than the IFA test. Seroprevalence, according to ELISA and IFA respectively, was higher in the aborting (88% and 82%) than in the non-aborting group (60% and 50%). High levels of serum antibodies were detected in goats post-abortion with an average of 114 %OD using ELISA and a log10(titer) of 2.4 using IFA. Strongly positive ELISA (%OD>80) and positive IFA results (log10(titers)>1.9) were significantly associated with abortion. Sampling on D15 gave the best association with ORs of 10 for ELISA and 6 for IFA. The practical interest of these results is discussed.
Assuntos
Anticorpos Antibacterianos/sangue , Coxiella burnetii/imunologia , Doenças das Cabras/diagnóstico , Febre Q/veterinária , Aborto Animal/diagnóstico , Aborto Animal/microbiologia , Animais , Antígenos de Bactérias/imunologia , Testes de Fixação de Complemento/métodos , Testes de Fixação de Complemento/normas , Testes de Fixação de Complemento/veterinária , Coxiella burnetii/isolamento & purificação , Diagnóstico Diferencial , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática/normas , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Técnica Indireta de Fluorescência para Anticorpo/métodos , Técnica Indireta de Fluorescência para Anticorpo/normas , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Doenças das Cabras/epidemiologia , Cabras , Gravidez , Febre Q/diagnóstico , Febre Q/epidemiologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Estudos SoroepidemiológicosRESUMO
Information on single-nucleotide polymorphisms (SNPs) in hexaploid bread wheat is still scarce. The goal of this study was to detect SNPs in wheat and examine their frequency. Twenty-six bread wheat lines from different origins worldwide were used. Specific PCR-products were obtained from 21 genes and directly sequenced. SNPs were discovered from the alignment of these sequences. The overall sequence polymorphism observed in this sample appears to be low; 64 single-base polymorphisms were detected in approximately 21.5 kb (i.e., 1 SNP every 335 bp). The level of polymorphism is highly variable among the different genes studied. Fifty percent of the genes studied contained no sequence polymorphism, whereas most SNPs detected were located in only 2 genes. As expected, taking into account a synthetic line created with a wild Triticum tauschii parent increases the level of polymorphism (101 SNPs; 1 SNP every 212 bp). The detected SNPs are available at http://urgi.versailles.inra.fr/GnpSNP">http://urgi.versailles.inra.fr/GnpSNP. Data on linkage disequilibrium (LD) are still preliminary. They showed a significant level of LD in the 2 most polymorphic genes. To conclude, the genome size of hexaploid wheat and its low level of polymorphism complicate SNP discovery in this species.