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1.
J Neurosci ; 44(2)2024 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-37968116

RESUMO

Reversal learning measures the ability to form flexible associations between choice outcomes with stimuli and actions that precede them. This type of learning is thought to rely on several cortical and subcortical areas, including the highly interconnected orbitofrontal cortex (OFC) and basolateral amygdala (BLA), and is often impaired in various neuropsychiatric and substance use disorders. However, the unique contributions of these regions to stimulus- and action-based reversal learning have not been systematically compared using a chemogenetic approach particularly before and after the first reversal that introduces new uncertainty. Here, we examined the roles of ventrolateral OFC (vlOFC) and BLA during reversal learning. Male and female rats were prepared with inhibitory designer receptors exclusively activated by designer drugs targeting projection neurons in these regions and tested on a series of deterministic and probabilistic reversals during which they learned about stimulus identity or side (left or right) associated with different reward probabilities. Using a counterbalanced within-subject design, we inhibited these regions prior to reversal sessions. We assessed initial and pre-/post-reversal changes in performance to measure learning and adjustments to reversals, respectively. We found that inhibition of the ventrolateral orbitofrontal cortex (vlOFC), but not BLA, eliminated adjustments to stimulus-based reversals. Inhibition of BLA, but not vlOFC, selectively impaired action-based probabilistic reversal learning, leaving deterministic reversal learning intact. vlOFC exhibited a sex-dependent role in early adjustment to action-based reversals, but not in overall learning. These results reveal dissociable roles for BLA and vlOFC in flexible learning and highlight a more crucial role for BLA in learning meaningful changes in the reward environment.


Assuntos
Complexo Nuclear Basolateral da Amígdala , Ratos , Masculino , Feminino , Animais , Incerteza , Complexo Nuclear Basolateral da Amígdala/fisiologia , Ratos Long-Evans , Córtex Pré-Frontal/fisiologia , Reversão de Aprendizagem/fisiologia
2.
Nanotechnology ; 32(41)2021 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-34285145

RESUMO

This study focuses on the development of a new hybrid biological material to be applied in the production of electrical energy. These organo-metallic cells are constituted by cyanobacteria (Fischerella muscicola) and silver nanoparticles (AgNPs). AgNPs were obtained by green synthesis using the extract of the fruit of theBerberis halliiplant as reducing agent with two different concentrations of silver nitrate (AgNO3), 1 and 10 mM. The morphology, physicochemical and electrical properties of the cyanobacteria with and without AgNPs were evaluated. To verify the efficacy of this new material, and the effect of the medium used, Nitrofoska or BG-11, the growth kinetics was evaluated by UV-vis up tot= 63 d with and without renewal of the culture medium and O2/CO2exchange. Through morphological characterizations ofFischerella muscicolait was possible to identify the presence of an associated bacterium identified using molecular techniques asPseudomona guguanensithat could act as a supporting organism in the growth of this cyanobacteria. The studies carried out did not shown cell toxicity for the cultures that have AgNPs and on the other hand, it was observed that the hybrid cells (Cy-AgNPs) are electron carriers recording an increase of up to 57% and 18% in their electrical potential with BG-11 and Nitrofoska culture media, respectively and an increase in the anodic current peak of 6.5% of Cy-AgNPs respect to onlyF. musicola.

3.
J Helminthol ; 95: e46, 2021 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-34412711

RESUMO

The objective of this study is to determine the prevalence of Fasciola hepatica infection in cattle slaughterhouses, as well as its association with climatic/environmental factors (derived from satellite data), seasonality and climate regions in two states in Mexico. Condemned livers from slaughtered animals were obtained from three abattoirs in the states of Puebla and Veracruz. The overall prevalence of the parasite in cattle between January and December of 2017 was 20.6% (1407 out of 6834); the highest rate of condemnation was observed in Veracruz (26.3%; tropical climate), and the lowest rate was found in Puebla (15.5%; temperate climate). The seasonal prevalence of fluke infection was 18.6%, 14.8% and 28.4% during the wet season, and 17.1%, 12.4% and 22.8% during the dry season in the three abattoir sites, located in the districts of Zacatlán, Teziutlán and Ciudad Alemán, respectively. Liver condemnations due to bovine fasciolosis were prevalent in the Zacatlán, Teziutlán and Ciudad Alemán districts during summer, autumn and summer, respectively. Using generalized estimating equations analysis, we determined six variables - rainfall (wet/dry), land surface temperature day, land surface temperature night, normalized difference vegetation index, seasonality and climate regions (temperate/tropical) - to be significantly associated with the prevalence of condemned livers. Climate region was the variable most strongly associated with F. hepatica infection (odds ratio (OR) 266.59; 95% confidence interval (CI): 241.90-353.34), followed by wet and dry seasons (OR 25.56; 95% CI: 20.56-55.67).


Assuntos
Doenças dos Bovinos , Fasciola hepatica , Fasciolíase , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Fasciolíase/epidemiologia , Fasciolíase/veterinária , México/epidemiologia , Prevalência , Estações do Ano , Clima Tropical
4.
Eur J Pediatr ; 179(8): 1279-1285, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32504135

RESUMO

Since March 2020, the world is involved in the COVID-19 pandemic, a disease caused by a novel virus called SARS-CoV-2. Some authors have described the ultrasonographic findings of COVID-19 pneumonia in adults and children, but data on neonates are lacking. Our objective was to describe the ultrasonographic lung pattern on newborns with SARS-CoV-2 infection during the COVID-19 pandemic. Newborns who tested positive for SARS-CoV-2 PCR in respiratory samples and were evaluated with point-of-care lung ultrasound (LU) from March to April 2020 were included. LU was performed bedside by a single investigator at the time of diagnosis and every 48 h during the first week following diagnosis. Six areas were studied. Three neonates were included. Infants' comorbidities included meconium aspiration syndrome, bronchopulmonary dysplasia, and Hirschsprung's disease. One required mechanical ventilation. No deaths occurred. LU showed B-lines, consolidation, and spared areas. No pneumothorax or pleural effusion was observedConclusions: LU could be of value when managing COVID-19 neonates. We describe the findings of lung ultrasound monitoring during the first week following diagnosis in three neonates with SARS-CoV-2 infection. What is known: • Lung ultrasound (LU) is a useful tool in COVID-19 management in adults. To date, no report on LU and neonates with SARS-CoV-2 infection has been published. What is new: • This study adds evidence about LU findings in neonates with SARS-CoV-2 infection.


Assuntos
Betacoronavirus , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/diagnóstico por imagem , Pulmão/diagnóstico por imagem , Pneumonia Viral/diagnóstico por imagem , Sistemas Automatizados de Assistência Junto ao Leito , COVID-19 , Teste para COVID-19 , Infecções por Coronavirus/complicações , Infecções por Coronavirus/diagnóstico , Humanos , Recém-Nascido , Masculino , Pandemias , Pneumonia Viral/complicações , SARS-CoV-2 , Ultrassonografia
5.
Persoonia ; 44: 67-97, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33116336

RESUMO

Mucor species are common soil fungi but also known as agents of human infections (mucormycosis) and used in food production and biotechnology. Mucor circinelloides is the Mucor species that is most frequently isolated from clinical sources. The taxonomy of Mucor circinelloides and its close relatives (Mucor circinelloides complex - MCC) is still based on morphology and mating behaviour. The aim of the present study was a revised taxonomy of the MCC using a polyphasic approach. Using a set of 100 strains molecular phylogenetic analysis of five markers (ITS, rpb1, tsr1, mcm7, and cfs, introduced here) were performed, combined with phenotypic studies, mating tests and the determination of the maximum growth temperatures. The multi-locus analyses revealed 16 phylogenetic species of which 14 showed distinct phenotypical traits and were recognised as discrete species. Five of these species are introduced as novel taxa: M. amethystinus sp. nov., M. atramentarius sp. nov., M. variicolumellatus sp. nov., M. pseudocircinelloides sp. nov., and M. pseudolusitanicus sp. nov. The former formae of M. circinelloides represent one or two separate species. In the MCC, the simple presence of well-shaped zygospores only indicates a close relation of both strains, but not necessarily conspecificity. Seven species of the MCC have been implemented in human infection: M. circinelloides, M. griseocyanus, M. janssenii, M. lusitanicus, M. ramosissimus, M. variicolumellatus, and M. velutinosus.

6.
Artigo em Inglês | MEDLINE | ID: mdl-30323038

RESUMO

Although the Sensititre Yeast-One (SYO) and Etest methods are widely utilized, interpretive criteria are not available for triazole susceptibility testing of Candida or Aspergillus species. We collected fluconazole, itraconazole, posaconazole, and voriconazole SYO and Etest MICs from 39 laboratories representing all continents for (method/agent-dependent) 11,171 Candida albicans, 215 C. dubliniensis, 4,418 C. glabrata species complex, 157 C.guilliermondii (Meyerozyma guilliermondii), 676 C. krusei (Pichia kudriavzevii), 298 C.lusitaniae (Clavispora lusitaniae), 911 C.parapsilosissensu stricto, 3,691 C.parapsilosis species complex, 36 C.metapsilosis, 110 C.orthopsilosis, 1,854 C.tropicalis, 244 Saccharomyces cerevisiae, 1,409 Aspergillus fumigatus, 389 A.flavus, 130 A.nidulans, 233 A.niger, and 302 A.terreus complex isolates. SYO/Etest MICs for 282 confirmed non-wild-type (non-WT) isolates were included: ERG11 (C. albicans), ERG11 and MRR1 (C. parapsilosis), cyp51A (A. fumigatus), and CDR2 and CDR1 overexpression (C. albicans and C. glabrata, respectively). Interlaboratory modal agreement was superior by SYO for yeast species and by the Etest for Aspergillus spp. Distributions fulfilling CLSI criteria for epidemiological cutoff value (ECV) definition were pooled, and we proposed SYO ECVs for S. cerevisiae and 9 yeast and 3 Aspergillus species and Etest ECVs for 5 yeast and 4 Aspergillus species. The posaconazole SYO ECV of 0.06 µg/ml for C. albicans and the Etest itraconazole ECV of 2 µg/ml for A. fumigatus were the best predictors of non-WT isolates. These findings support the need for method-dependent ECVs, as, overall, the SYO appears to perform better for susceptibility testing of yeast species and the Etest appears to perform better for susceptibility testing of Aspergillus spp. Further evaluations should be conducted with more Candida mutants.


Assuntos
Antifúngicos/farmacologia , Aspergillus/efeitos dos fármacos , Candida/efeitos dos fármacos , Triazóis/farmacologia , Aspergilose/tratamento farmacológico , Aspergilose/epidemiologia , Aspergilose/microbiologia , Aspergillus/classificação , Aspergillus/isolamento & purificação , Candida/classificação , Candida/isolamento & purificação , Candidíase/tratamento farmacológico , Candidíase/epidemiologia , Candidíase/microbiologia , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Farmacorresistência Fúngica , Fluconazol/farmacologia , Humanos , Hospedeiro Imunocomprometido , Itraconazol/farmacologia , Voriconazol/farmacologia
7.
Eur Arch Otorhinolaryngol ; 276(11): 3095-3104, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31363901

RESUMO

PURPOSE: Intense bleeding of the surgical field is a potential factor influencing success of functional endoscopic sinus surgery (FESS). Hypotensive anesthesia with α2 intravenous agonists reduces intraoperative bleeding, but which is the best agent is unknown. The main objective of this trial was to compare the current standard adjuvant drug for hypotensive anesthesia, clonidine, with the recently available alternative dexmedetomidine. METHODS: A randomized clinical trial compared the efficacy of clonidine and dexmedetomidine during FESS. Treatment was open label for the anesthesiologist and operating surgeon, but blind for an external evaluator who evaluated video-recorded surgeries. A Boezaart scale was assessed every 30 min during FESS until surgery completion. Main end-point was the proportion of patients with mean Boezaart scores > 2 (heavy bleeding) by external blinded evaluator. Secondary end-points included other bleeding parameters, surgery duration, hemodynamic measures and surgical complications. RESULTS: 94 patients were randomized. There were no significant differences in the proportion of patients with mean Boezaart scores > 2 in clonidine (42.6%) and dexmedetomidine (42.6%). Consistently, no differences were observed in secondary variables of bleeding, duration or complications. Small differences in mean heart rate were observed that might reflect different pharmacological profiles of the products, but are of uncertain clinical relevance. CONCLUSIONS: No significant differences were observed between clonidine and dexmedetomidine when used as anesthetic adjuvants in the reduction of surgical bleeding in FESS. A longer experience with clonidine and its lower costs suggest it may be a preferable option as an adjuvant for hypotensive anesthesia.


Assuntos
Anestesia/métodos , Perda Sanguínea Cirúrgica/prevenção & controle , Clonidina , Dexmedetomidina , Endoscopia , Seios Paranasais/cirurgia , Agonistas de Receptores Adrenérgicos alfa 2/administração & dosagem , Agonistas de Receptores Adrenérgicos alfa 2/efeitos adversos , Doença Crônica , Clonidina/administração & dosagem , Clonidina/efeitos adversos , Dexmedetomidina/administração & dosagem , Dexmedetomidina/efeitos adversos , Endoscopia/efeitos adversos , Endoscopia/métodos , Feminino , Hemodinâmica/efeitos dos fármacos , Humanos , Masculino , Pessoa de Meia-Idade , Pólipos Nasais/cirurgia , Duração da Cirurgia , Rinite/cirurgia , Sinusite/cirurgia , Resultado do Tratamento
8.
Rhinology ; 57(4): 252-260, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-30928998

RESUMO

BACKGROUND: Macrolide antibiotics have demonstrated important anti-inflammatory and immunomodulatory properties in chronic rhinosinusitis (CRS) patients. However, reports of increased risks of cardiovascular events have led to safety concerns. We investigated the risk of all-cause and cardiac death, and cardiovascular outcomes, associated with macrolide use. METHODOLOGY: Observational cohort (1997-2016) using linked data from the Clinical Practice Research Datalink, Hospital Episodes Statistics, and the Office for National Statistics. Patients aged 16-80 years with CRS prescribed a macrolide antibiotic or penicillin were included, comparing prescriptions for macrolide antibiotics to penicillin. Outcomes were all-cause mortality, cardiac death, myocardial infarction, stroke, diagnosis of peripheral vascular disease, and cardiac arrhythmia. RESULTS: Analysis included 320,798 prescriptions received by 66,331 patients. There were 3,251 deaths, 815 due to cardiovascular causes, 925 incident myocardial infarctions, 859 strokes, 637 diagnoses of peripheral vascular disease, and 1,436 cardiac arrhythmias. A non-statistically significant trend towards increased risk of myocardial infarction during the first 30 days following macrolide prescription was observed. No statistically significant short- or long-term risks were observed for macrolide prescription. No significant risks were identified for clarithromycin in particular. CONCLUSIONS: Although not statistically significant, our best estimates suggest an increased short-term risk of myocardial infarction in patients with CRS following macrolide prescription, supporting previous observational evidence. However, confounding by indication remains a possible explanation for this apparent increased risk. We found no evidence of longer term increased risks.


Assuntos
Antibacterianos , Doenças Cardiovasculares , Macrolídeos , Rinite , Sinusite , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/efeitos adversos , Antibacterianos/uso terapêutico , Doenças Cardiovasculares/epidemiologia , Estudos de Coortes , Registros Eletrônicos de Saúde , Humanos , Macrolídeos/efeitos adversos , Macrolídeos/uso terapêutico , Pessoa de Meia-Idade , Atenção Primária à Saúde , Rinite/tratamento farmacológico , Sinusite/tratamento farmacológico , Adulto Jovem
9.
Rev Med Chil ; 147(1): 34-40, 2019.
Artigo em Espanhol | MEDLINE | ID: mdl-30848762

RESUMO

BACKGROUND: Perioperative cardiac arrest (PCA) is a rare but important event in the operating room. AIM: To describe PCA events at a Clinical Hospital in Santiago, Chile. MATERIAL AND METHODS: Registry of PCA that occurred in the operating room (OR) and during procedures not carried out in the OR between September 2006 and November 2017. Precipitating events, type of anesthesia and results of resuscitation maneuvers were described. RESULTS: Eighty events (five outside of the OR) during 170,431 surgical procedures were recorded, resulting in an incidence of 4.4 events per 10,000 interventions. Hypotension/hypoperfusion was the most frequently found preexisting condition (42.5%). The main cause was the presence of preoperative complications (57.5%). Nineteen cases (23.8%) were attributable to anesthesia, with an incidence of 1.11 per 10,000 anesthetic procedures. Survival rate at hospital discharge was 52.5%. The figure for PCA caused by anesthesia was 84.2%. CONCLUSIONS: The incidence of PCA and its survival is similar to that reported abroad. In general, PCA has a better prognosis than other types of cardiac arrest, especially if it has an anesthetic cause.


Assuntos
Parada Cardíaca/epidemiologia , Hospitais Universitários/estatística & dados numéricos , Complicações Intraoperatórias/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anestesia/efeitos adversos , Anestesia/estatística & dados numéricos , Criança , Pré-Escolar , Chile/epidemiologia , Feminino , Parada Cardíaca/etiologia , Mortalidade Hospitalar , Humanos , Incidência , Lactente , Complicações Intraoperatórias/etiologia , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Taxa de Sobrevida , Fatores de Tempo , Adulto Jovem
10.
Artigo em Inglês | MEDLINE | ID: mdl-29437624

RESUMO

Estimating epidemiological cutoff endpoints (ECVs/ECOFFS) may be hindered by the overlap of MICs for mutant and nonmutant strains (strains harboring or not harboring mutations, respectively). Posaconazole MIC distributions for the Aspergillus fumigatus species complex were collected from 26 laboratories (in Australia, Canada, Europe, India, South and North America, and Taiwan) and published studies. Distributions that fulfilled CLSI criteria were pooled and ECVs were estimated. The sensitivity of three ECV analytical techniques (the ECOFFinder, normalized resistance interpretation [NRI], derivatization methods) to the inclusion of MICs for mutants was examined for three susceptibility testing methods (the CLSI, EUCAST, and Etest methods). The totals of posaconazole MICs for nonmutant isolates (isolates with no known cyp51A mutations) and mutant A. fumigatus isolates were as follows: by the CLSI method, 2,223 and 274, respectively; by the EUCAST method, 556 and 52, respectively; and by Etest, 1,365 and 29, respectively. MICs for 381 isolates with unknown mutational status were also evaluated with the Sensititre YeastOne system (SYO). We observed an overlap in posaconazole MICs among nonmutants and cyp51A mutants. At the commonly chosen percentage of the modeled wild-type population (97.5%), almost all ECVs remained the same when the MICs for nonmutant and mutant distributions were merged: ECOFFinder ECVs, 0.5 µg/ml for the CLSI method and 0.25 µg/ml for the EUCAST method and Etest; NRI ECVs, 0.5 µg/ml for all three methods. However, the ECOFFinder ECV for 95% of the nonmutant population by the CLSI method was 0.25 µg/ml. The tentative ECOFFinder ECV with SYO was 0.06 µg/ml (data from 3/8 laboratories). Derivatization ECVs with or without mutant inclusion were either 0.25 µg/ml (CLSI, EUCAST, Etest) or 0.06 µg/ml (SYO). It appears that ECV analytical techniques may not be vulnerable to overlap between presumptive wild-type isolates and cyp51A mutants when up to 11.6% of the estimated wild-type population includes mutants.


Assuntos
Antifúngicos/farmacologia , Aspergillus fumigatus/efeitos dos fármacos , Aspergillus fumigatus/genética , Mutação/genética , Triazóis/farmacologia , Farmacorresistência Fúngica/genética , Testes de Sensibilidade Microbiana , Voriconazol/farmacologia
11.
Med Mycol ; 56(1): 51-59, 2018 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-28431110

RESUMO

African histoplasmosis is defined as the fungal infection caused by Histoplasma capsulatum var. duboisii (Hcd). Studies focused on distinguishing Hcd and H. capsulatum var. capsulatum (Hcc), which coexist in Africa, are scarce or outdated, and African strains are continuously underrepresented. In this work, 13 cases of African patients with histoplasmosis diagnosed in the Spanish Mycology Reference Laboratory have been reviewed showing that 77% had disseminated disease and AIDS as underlying disease although Hcd infection has been classically considered a rare presentation in AIDS patients. Strains isolated from these patients and other clinical and reference strains were studied by assessing classical identification methods and performing a three-loci multi-locus sequence analysis (MLSA). Classical identification methods based on biochemical tests and measurement of yeast size proved to be useless in distinguishing both varieties. The MLSA defined an African cluster, with a strong statistical support, that included all strains with African origin. Finally, mating type was also determined by using molecular methods revealing an unequal mating type distribution in African strains. In conclusion, historical statements and classical identification methods were useless to distinguish between varieties, whereas molecular analyses revealed that all strains with African origin grouped together suggesting that traditional classification should be revised. Further investigation is required in order to unravel traditional concepts about Hcd infection and support results obtained in this work.


Assuntos
Histoplasma/classificação , Histoplasma/isolamento & purificação , Histoplasmose/microbiologia , Histoplasmose/patologia , Infecções Oportunistas Relacionadas com a AIDS/microbiologia , Infecções Oportunistas Relacionadas com a AIDS/patologia , Adulto , Idoso , Feminino , Genes Fúngicos Tipo Acasalamento , Genótipo , Histoplasma/genética , Histoplasma/fisiologia , Humanos , Infecções Fúngicas Invasivas/microbiologia , Infecções Fúngicas Invasivas/patologia , Masculino , Pessoa de Meia-Idade , Tipagem de Sequências Multilocus , Técnicas de Tipagem Micológica , Espanha , Adulto Jovem
12.
Mycopathologia ; 183(1): 185-200, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28929280

RESUMO

The genus Scedosporium, which comprises at least five clinically relevant species, i.e. Scedosporium apiospermum, Scedosporium boydii, Scedosporium aurantiacum, Scedosporium dehoogii and Scedosporium minutisporum, ranks the second among the filamentous fungi colonizing the airways of patients with cystic fibrosis (CF). This colonization of the airways is thought to contribute to the inflammatory reaction leading to a progressive deterioration of the lung function. Additionally, these colonizing fungi may lead to severe disseminated infections in case of lung transplantation. Therefore, considering the low susceptibility of Scedosporium species to all current antifungal drugs, preventive measures should be defined to reduce the risk of exposure to these fungi for non-colonized CF patients. With this in mind, several studies have been conducted to elucidate the ecology of these fungi and to define possible sources of patient contamination. This review will summarize the major outcomes of those studies, including: the clear demonstration that ecological niches of Scedosporium species are strongly impacted by human activities, and the ability of Scedosporium species to degrade aliphatic and aromatic pollutants which supports the high occurrence of these species in contaminated soils and polluted waters and makes them promising candidates for bioremediation purposes. Finally, prospects for future research in this field are proposed.


Assuntos
Fibrose Cística/complicações , Fibrose Cística/microbiologia , Pneumopatias Fúngicas/microbiologia , Scedosporium/crescimento & desenvolvimento , Scedosporium/isolamento & purificação , Exposição Ambiental , Humanos , Scedosporium/classificação
13.
Artigo em Inglês | MEDLINE | ID: mdl-28893791

RESUMO

The global emergence of azole-resistant Aspergillus fumigatus strains is a growing public health concern. Different patterns of azole resistance are linked to mutations in cyp51A Therefore, accurate characterization of the mechanisms underlying azole resistance is critical to guide selection of the most appropriate antifungal agent for patients with aspergillosis. This study describes a new sequencing-free molecular screening tool for early detection of the most frequent mutations known to be associated with azole resistance in A. fumigatus PCRs targeting cyp51A mutations at positions G54, Y121, G448, and M220 and targeting different tandem repeats (TRs) in the promoter region were designed. All PCRs were performed simultaneously, using the same cycling conditions. Amplicons were then distinguished using a high-resolution melting assay. For standardization, 30 well-characterized azole-resistant A. fumigatus strains were used, yielding melting curve clusters for different resistance mechanisms for each target and allowing detection of the most frequent azole resistance mutations, i.e., G54E, G54V, G54R, G54W, Y121F, M220V, M220I, M220T, M220K, and G448S, and the tandem repeats TR34, TR46, and TR53 Validation of the method was performed using a blind panel of 80 A. fumigatus azole-susceptible or azole-resistant strains. All strains included in the blind panel were properly classified as susceptible or resistant with the developed method. The implementation of this screening method can reduce the time needed for the detection of azole-resistant A. fumigatus isolates and therefore facilitate selection of the best antifungal therapy in patients with aspergillosis.


Assuntos
Antifúngicos/farmacologia , Aspergillus fumigatus/efeitos dos fármacos , Sistema Enzimático do Citocromo P-450/genética , Farmacorresistência Fúngica/genética , Proteínas Fúngicas/genética , Reação em Cadeia da Polimerase/métodos , Aspergilose/tratamento farmacológico , Aspergillus fumigatus/genética , Humanos , Testes de Sensibilidade Microbiana , Desnaturação de Ácido Nucleico/genética , Regiões Promotoras Genéticas/genética , Sequências de Repetição em Tandem/genética
14.
J Microsc ; 267(2): 130-142, 2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-28319261

RESUMO

Measurement of collagen bundle orientation in histopathological samples is a widely used and useful technique in many research and clinical scenarios. Fourier analysis is the preferred method for performing this measurement, but the most appropriate staining and microscopy technique remains unclear. Some authors advocate the use of Haematoxylin-Eosin (H&E) and confocal microscopy, but there are no studies comparing this technique with other classical collagen stainings. In our study, 46 human skin samples were collected, processed for histological analysis and stained with Masson's trichrome, Picrosirius red and H&E. Five microphotographs of the reticular dermis were taken with a 200× magnification with light microscopy, polarized microscopy and confocal microscopy, respectively. Two independent observers measured collagen bundle orientation with semiautomated Fourier analysis with the Image-Pro Plus 7.0 software and three independent observers performed a semiquantitative evaluation of the same parameter. The average orientation for each case was calculated with the values of the five pictures. We analyzed the interrater reliability, the consistency between Fourier analysis and average semiquantitative evaluation and the consistency between measurements in Masson's trichrome, Picrosirius red and H&E-confocal. Statistical analysis for reliability and agreement was performed with the SPSS 22.0 software and consisted of intraclass correlation coefficient (ICC), Bland-Altman plots and limits of agreement and coefficient of variation. Interrater reliability was almost perfect (ICC > 0.8) with all three histological and microscopy techniques and always superior in Fourier analysis than in average semiquantitative evaluation. Measurements were consistent between Fourier analysis by one observer and average semiquantitative evaluation by three observers, with an almost perfect agreement with Masson's trichrome and Picrosirius red techniques (ICC > 0.8) and a strong agreement with H&E-confocal (0.7 < ICC < 0.8). Comparison of measurements between the three techniques for the same observer showed an almost perfect agreement (ICC > 0.8), better with Fourier analysis than with semiquantitative evaluation (single and average). These results in nonpathological skin samples were also confirmed in a preliminary analysis in eight scleroderma skin samples. Our results show that Masson's trichrome and Picrosirius red are consistent with H&E-confocal for measuring collagen bundle orientation in histological samples and could thus be used indistinctly for this purpose. Fourier analysis is superior to average semiquantitative evaluation and should keep being used as the preferred method.


Assuntos
Colágeno/análise , Análise de Fourier , Histocitoquímica/métodos , Microscopia Confocal/métodos , Microscopia de Polarização/métodos , Microscopia/métodos , Coloração e Rotulagem/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Feminino , Hospitais Universitários , Humanos , Processamento de Imagem Assistida por Computador/métodos , Lactente , Masculino , Pessoa de Meia-Idade , Pele/patologia , Espanha , Adulto Jovem
15.
Clin Genet ; 90(4): 361-5, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-26864382

RESUMO

Breast cancer (BC) is the most frequent cancer among women in Morocco. However, the role of the most prevalent BC-predisposing genes, BRCA1 and BRCA2, has been largely unexplored. To help define the role of BRCA1 in BC in Morocco, we characterized the first potential BRCA1 founder mutation in this population. Genetic testing of BRCA1 and BRCA2 in BC high-risk families identified mutation BRCA1 c.5309G>T, p.(Gly1770Val) or G1770V in five independent families from Morocco, suggesting a founder effect. To confirm this hypothesis, haplotype construction was performed using seven intragenic and flanking BRCA1 microsatellite markers. Clinical data were also compiled. Clinical data from carriers of mutation G1770V correspond to data from carriers of BRCA1 pathogenic mutations. Microsatellite analysis showed a common haplotype for the five families in a region comprising 1.54 Mb, confirming G1770V as the first specific founder BRCA1 mutation in the Moroccan population. Our findings contribute to a better understanding of BC genetics in the Moroccan population. Nevertheless, comprehensive studies of mutation G1770V in large series of BC patients from Morocco are needed to assess the real prevalence of this mutation and to improve genetic testing and risk assessment in this population.


Assuntos
Proteína BRCA1/genética , Efeito Fundador , Mutação , Adulto , Proteína BRCA1/química , Feminino , Predisposição Genética para Doença , Haplótipos , Humanos , Repetições de Microssatélites , Pessoa de Meia-Idade , Marrocos , Linhagem
16.
Antimicrob Agents Chemother ; 59(11): 6975-82, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26324281

RESUMO

In invasive candidiasis, there has been an epidemiological shift from Candida albicans to non-albicans species infections, including infections with C. glabrata, C. parapsilosis, C. tropicalis, and C. krusei. Although the prevalence of C. krusei remains low among yeast infections, its intrinsic resistance to fluconazole raises epidemiological and therapeutic concerns. Echinocandins have in vitro activity against most Candida spp. and are the first-line agents in the treatment of candidemia. Although resistance to echinocandin drugs is still rare, individual cases of C. krusei resistance have been reported in recent years, especially with strains that have been under selective pressure. A total of 15 C. krusei strains, isolated from the blood, urine, and soft tissue of an acute lymphocytic leukemia patient, were analyzed. Strains developed echinocandin resistance during 10 days of caspofungin therapy. The molecular epidemiology of the isolates was investigated using two different typing methods: PCR-based amplification of the species-specific repetitive polymorphic CKRS-1 sequence and multilocus sequence typing. All isolates were genetically related, and the mechanism involved in decreased echinocandin susceptibility was characterized. Clinical resistance was associated with an increase in echinocandin MICs in vitro and was related to three different mutations in hot spot 1 of the target enzyme Fks1p. Molecular evidence of the rapid acquisition of resistance by different mutations in FKS1 highlights the need to monitor the development of resistance in C. krusei infections treated with echinocandin drugs.


Assuntos
Antifúngicos/farmacologia , Candida/efeitos dos fármacos , Equinocandinas/farmacologia , Antifúngicos/uso terapêutico , Candida/genética , Candida/patogenicidade , Candidíase/tratamento farmacológico , Candidíase/microbiologia , Caspofungina , Equinocandinas/uso terapêutico , Feminino , Humanos , Lipopeptídeos , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase
17.
Child Care Health Dev ; 41(2): 186-93, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24635011

RESUMO

BACKGROUND: Consideration of child safeguarding is routine within maternity services but less common in other health services for adults. We audited notifications for child safeguarding from an acute general hospital where the policy includes questioning adults presenting with violence, mental health problems or drug or alcohol misuse to any department within the hospital about children at home and notifying to the local authority children's social care services if there are safeguarding concerns. METHODS: Cross-sectional audit of notifications for child safeguarding, including abuse, neglect or victimization, from all departments in one hospital to the local authority children's social care department during 12 months (2010/11). RESULTS: Of 681 notifications (57 per month), 40% (270/681) were triggered by parents' presentation to acute hospital services. Of these, 37% (100/270; 12 teenage mothers) presented for maternity care and 60% (162/270; 8 teenage parents) presented to the emergency department (ED). Of the 60% (411/681) of notifications prompted by children presenting for healthcare, most originated from the ED (358/411; 87%): two-thirds of these presented with injury (250/358; 70%). CONCLUSION: Given a policy to ask adults about children at home, a substantial proportion of children notified for child safeguarding were recognized through presentations to acute healthcare by their parents. Further research and development of this policy needs to ensure that questioning results in effective interventions for the children and their parents.


Assuntos
Maus-Tratos Infantis/diagnóstico , Filho de Pais com Deficiência/psicologia , Hospitais Gerais/organização & administração , Pais/psicologia , Serviço Social/organização & administração , Adolescente , Criança , Maus-Tratos Infantis/prevenção & controle , Proteção da Criança , Estudos Transversais , Serviço Hospitalar de Emergência/organização & administração , Saúde da Família , Feminino , Humanos , Londres , Masculino , Encaminhamento e Consulta/estatística & dados numéricos , Serviço Social/estatística & dados numéricos
18.
Arch Environ Contam Toxicol ; 68(1): 181-92, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25015731

RESUMO

Bioavailability is affected by soil physicochemical characteristics such as pH and organic matter (OM) content. In addition, OM constitutes the energy source of Eisenia fetida, a well established model species for soil toxicity assessment. The present work aimed at assessing the effects of changes in OM content on the toxicity of Cd in E. fetida through the measurement of neutral red uptake (NRU) and mortality, growth, and reproduction (Organisation for Economic Co-operation and Development [OECD] Nos. 207 and 222). Complementarily, metallothionein (MT) and catalase transcription levels were measured. To decrease variability inherent to natural soils, artificial soils (Organization for Economic Cooperation and Development 1984) with different OM content (6, 10, and 14%) and spiked with Cd solutions at increasing concentrations were used. Low OM in soil decreased soil ingestion and Cd bioaccumulation but also increased Cd toxicity causing lower NRU of coelomocytes, 100 % mortality, and stronger reproduction impairment, probably due to the lack of energy to maintain protection mechanisms (production of MT).Cd bioaccumulation did not reflect toxicity, and OM played a pivotal role in Cd toxicity. Thus, OM content should be taken into account when using E. fetida in in vivo exposures for soil health assessment.


Assuntos
Cádmio/toxicidade , Poluentes do Solo/toxicidade , Solo/química , Animais , Biomarcadores , Metalotioneína , Oligoquetos , Testes de Toxicidade
19.
Plant Foods Hum Nutr ; 70(2): 153-9, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25736875

RESUMO

The fermentation of fruit produces significant changes in their nutritional composition. An orange beverage has been obtained from the controlled alcoholic fermentation and thermal pasteurization of orange juice. A study was performed to determine the influence of both processes on its amino acid profile. UHPLC-QqQ-MS/MS was used for the first time for analysis of orange juice samples. Out of 29 amino acids and derivatives identified, eight (ethanolamine, ornithine, phosphoethanolamine, α-amino-n-butyric acid, hydroxyproline, methylhistidine, citrulline, and cystathionine) have not previously been detected in orange juice. The amino acid profile of the orange juice was not modified by its processing, but total amino acid content of the juice (8194 mg/L) was significantly increased at 9 days of fermentation (13,324 mg/L). Although the pasteurization process produced partial amino acid degradation, the total amino acid content was higher in the final product (9265 mg/L) than in the original juice, enhancing its nutritional value.


Assuntos
Aminoácidos/análise , Citrus sinensis/química , Fermentação , Sucos de Frutas e Vegetais/análise , Pasteurização , Espectrometria de Massas em Tandem
20.
J Anat ; 225(1): 98-108, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24754576

RESUMO

The tissue organisation of dermal collagen is gaining importance as a contributing factor both in development and ageing, as well as in skin maturation processes. In this work we aim to study different representative parameters of this structural organisation in 45 human skin samples of assorted ages, by means of image analysis. The variation of these parameters on the basis of age was assessed using several regression models (linear, quadratic and cubic). The area occupied by collagen was significantly reduced as a function of age in the papillary dermis (R(2) = 0.437, P < 0.0001), as well as the thickness of the collagen bundles (R(2) = 0.461, P < 0.0001), following statistical models of cubic and quadratic regression, respectively. The width of the papillary dermis increased in a significant manner over a linear regression model (R(2) = 0.26, P < 0.0001). In the reticular dermis, the cubic regression indicated a significant decline (R(2) = 0.392, P = 0.002) of the area filled with collagen according to the age. Both collagen thickness and bundle orientation parameters fit a quadratic regression over the age in a significant way (R(2) = 0.433 and R(2) = 0.334, respectively, both P < 0.0001). The width of the reticular dermis followed also a significant quadratic distribution according to age (R(2) = 0.193, P = 0.011). These parameters could partially explain the lifelong functional changes taking place in the skin and propose a baseline providing a useful entry point for future investigation.


Assuntos
Colágeno/análise , Derme/citologia , Envelhecimento da Pele/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Feminino , Humanos , Processamento de Imagem Assistida por Computador , Lactente , Masculino , Pessoa de Meia-Idade , Análise de Regressão , Adulto Jovem
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