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OBJECTIVES: Performance evaluation of routine laboratory methods to determine the susceptibility of Enterobacterales urinary isolates to fosfomycin (oral administration) and mecillinam. METHODS: We collected 347 Enterobacterales isolates from monomicrobial midstream urine samples from women with significant bacteriuria and leukocyturia. Mostly non-Escherichia coli isolates (i.e. Klebsiella spp., Citrobacter koseri, Enterobacter cloacae complex and Proteus mirabilis) were included (nâ=â298). Performance of VITEK®2, ETEST®, and disc diffusion to determine fosfomycin and mecillinam susceptibility was evaluated following International Organization for Standardization (ISO) 20776-2:2021 (or 20776-2:2007 for disc diffusion) in comparison with the agar dilution reference method. RESULTS: For fosfomycin testing, VITEK®2 and ETEST® were close to reaching ISO requirements (essential agreement â≥â90%; bias â±30%) for C. koseri, E. coli and P. mirabilis. Categorical agreement (CA) and major error rates were acceptable for disc diffusion. Fosfomycin displayed lower activity against E. cloacae complex and Klebsiella spp., with MIC50 (minimum inhibitory concentration required to inhibit the growth of 50% of tested isolates) equal to the E. coli EUCAST breakpoint (8â mg/L). For these species, the three alternative techniques overestimated MICs and resistance, and did not meet performance criteria. For mecillinam testing of Enterobacterales isolates, apart from P. mirabilis, ETEST® nearly fulfilled ISO requirements, and CA rates were acceptable for disc diffusion. ISO criteria were reached for C. koseri and E. coli testing with VITEK®2, apart from too high rates of very major errors. For P. mirabilis, performances were unacceptable, whatever the routine method used. CONCLUSIONS: Commercially available tests may serve as alternatives to agar dilution to assess fosfomycin (oral) and mecillinam susceptibility of Enterobacterales urinary isolates, with important interspecies variabilities. Additional studies comprising more fosfomycin- and mecillinam-resistant isolates are needed to strengthen our conclusions.
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BACKGROUND: Monoallelic germline MBD4 pathogenic variants were recently reported to cause a predisposition to uveal melanoma, associated with a specific tumor mutational signature and good response to immunotherapy. Monoallelic tumor pathogenic variants have also been described in brain tumors, breast cancers, and myxofibrosarcomas, whereas biallelic germline MBD4 pathogenic variants have been involved in a recessive hereditary adenomatous polyposis and a specific type of acute myeloid leukemia. METHODS: We analyzed MBD4 for all patients with a diagnosis of uveal melanoma at Institut Curie since July 2021 and in the 3240 consecutive female probands explored at the Institut Curie for suspicion of predisposition to breast cancer between July 2021 and February 2023. RESULTS: We describe 25 families whose probands carry a monoallelic germline pathogenic variant in MBD4. Eighteen of these families presented with uveal melanoma (including a case patient with multiple uveal melanoma), and 7 families presented with breast cancer. Family histories showed the first familial case of uveal melanoma in monoallelic MBD4 pathogenic variant carriers and other various types of cancers in relatives, especially breast, renal, and colorectal tumors. CONCLUSIONS: Monoallelic MBD4 pathogenic variant may explain some cases of familial and multiple uveal melanoma as well as various cancer types, expanding the tumor spectrum of this predisposition. Further genetic testing in relatives combined with molecular tumor analyses will help define the tumor spectrum and estimate each tumor's risk.
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Neoplasias de la Mama , Melanoma , Neoplasias Cutáneas , Neoplasias de la Úvea , Humanos , Adulto , Femenino , Predisposición Genética a la Enfermedad , Melanoma/epidemiología , Melanoma/genética , Melanoma/patología , Neoplasias Cutáneas/epidemiología , Neoplasias Cutáneas/genética , Mutación de Línea Germinal , Neoplasias de la Mama/epidemiología , Neoplasias de la Mama/genética , Endodesoxirribonucleasas/genéticaRESUMEN
Mentoring is considered an evidence-based practice for violence prevention. This study presents a partial replication of the Take Charge! program implemented in partnership with Big Brothers Big Sisters of America (BBBS). One hundred and eighty-eight early adolescents (M age = 12.87; 61.17% male) who were treated for peer-related assault injury in two urban mid-Atlantic emergency departments were randomly assigned to receive a mentor from two BBBS affiliates. Mentors and organization staff were trained in the Take Charge! violence prevention curriculum, which had previously shown evidence of efficacy. Intent-to-treat analyses showed statistically significant improvements in conflict avoidance self-efficacy for the intervention group at 9 months and reductions in fighting at 21 months, but an increase in parental report of aggression at 9 months. Complier average causal effect models revealed evidence of an additional effect for reduced problem behavior at 21 months for intervention adolescents who received a mentor. No effects were found for youth-reported aggression, retaliatory attitudes, deviance acceptance, or commitment to learning. Sensitivity analyses suggested increased aggressive behavior for adolescents in the intervention group who did not receive a mentor (i.e., non-compliers). These findings extend the evidence-base for Take Charge! as a violence prevention curriculum for youth already engaged in violence to "real-world" implementation settings. However, they also suggest that challenges associated with providing youth with mentors can be consequential and that additional supports may be needed for these youth/parents. Clinical trials number: clinicaltrials.gov NCT01770873.
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Víctimas de Crimen , Tutoría , Adolescente , Masculino , Humanos , Niño , Femenino , Mentores , Violencia/prevención & control , AgresiónRESUMEN
BACKGROUND: DICER1 syndrome is an autosomal dominant inherited syndrome predisposing to various benign and malignant tumors, mainly occurring in children and young adults, requiring broad surveillance starting at birth with repeated irradiating imaging exams and sedations for young patients. It is caused by monoallelic germline pathogenic variants in the DICER1 gene. More than 90% of tumors bear an additional somatic DICER1 missense hotspot mutation, as a second hit, involving 1 of 6 codons clustered in exons 24 and 25. We designed and in vitro validated a drop-off droplet digital PCR (ddPCR) system to scan all DICER1 hotspot codons, allowing for a liquid biopsy test, an alternative to sedation and radiation exposure. METHODS: Three drop-off ddPCR assays were designed, with 2 TaqMan probes per assay, 1 complementary to the wild-type sequence of the region containing hotspots and another 1 used as a reference. Eight tumor-derived DNAs and 5 synthetic oligonucleotides bearing DICER1 hotspot mutations were tested. RESULTS: All tested mutations were detected, with a limit of detection ranging from 0.07% to 0.31% for codons p. E1705, p. D1709, and p. D1713 in exon 24 and from 0.06% to 0.15% for codons p. G1809, p. D1810, and p. E1813 in exon 25. CONCLUSIONS: The high sensitivity of this method is compatible with its use for plasma circulating tumor DNA (ctDNA) analysis for early tumor detection in DICER1 syndrome patients. It may reduce the need for radiation exposure and sedation in surveillance protocols and may also improve patient prognosis. Clinical trials are needed to evaluate ctDNA analysis in these patients.
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ADN Tumoral Circulante , Neoplasias , Niño , Codón , ARN Helicasas DEAD-box/genética , Humanos , Recién Nacido , Mutación , Neoplasias/genética , Reacción en Cadena de la Polimerasa/métodos , Ribonucleasa III/genética , Adulto JovenRESUMEN
DICER1 syndrome is a rare paediatric autosomal dominant inherited disorder predisposing to various benign and malignant tumours. It is caused by a germline pathogenic variant in DICER1, and the second hit for tumour development is usually a missense hotspot pathogenic variant in the DICER1 ribonuclease IIIb domain. While DICER1 predisposing variants account for about 60% of ovarian Sertoli-Leydig cell tumours, no DICER1-related testicular stromal tumours have been described. Here we report the first two cases of testicular stromal tumours in children carrying a DICER1 germline pathogenic variant: a case of Sertoli cell tumour and a case of Leydig cell tumour diagnosed at 2 and 12 years of age, respectively. A somatic DICER1 hotspot pathogenic variant was detected in the Sertoli cell tumour. This report extends the spectrum of DICER1-related tumours to include testicular Sertoli cell tumour and potentially testicular Leydig cell tumour. Diagnosis of a testicular Sertoli cell tumour should prompt DICER1 genetic testing so that patients with a DICER1 germline pathogenic variant can benefit from established surveillance guidelines. DICER1 genetic evaluation may be considered for testicular Leydig cell tumour. Our findings suggest that miRNA dysregulation underlies the aetiology of some testicular stromal tumours.
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Tumor de Células de Leydig , Síndromes Neoplásicos Hereditarios , Neoplasias Ováricas , Tumor de Células de Sertoli , Tumor de Células de Sertoli-Leydig , Neoplasias Testiculares , Niño , ARN Helicasas DEAD-box/genética , Femenino , Humanos , Tumor de Células de Leydig/diagnóstico , Tumor de Células de Leydig/genética , Masculino , Neoplasias Ováricas/genética , Ribonucleasa III/genética , Tumor de Células de Sertoli/genética , Tumor de Células de Sertoli-Leydig/genética , Tumor de Células de Sertoli-Leydig/patología , Neoplasias Testiculares/genéticaRESUMEN
BACKGROUND: The neurological effects of Lyme borreliosis in children are varied and their clinical progression is not widely reported in the French literature. We carried out a retrospective study to describe the clinical characteristics of Lyme neuroborreliosis in children in southwest France and their clinical progression at 6 months. METHODS: This study was carried out at Toulouse University Hospital during the period 2006-2017 using patient records. Case definition was based on the combined French clinical and laboratory diagnostic criteria. RESULTS: In total, 26 children were included. The median age was 8 years (4-14 years). The different neurological symptoms reported were: meningoradiculitis (62%), which was usually associated with facial palsy (54%); isolated facial palsy (15%); isolated meningitis (8%); polyradiculoneuritis (4%); benign intracranial hypertension (4%) and myelomeningoradiculitis (4%). The most common functional symptoms were headaches (54%), the perception of asthenia (42%), neck pain (27%), and a loss of appetite (19%). Patients with laboratory meningitis (84%) often had no signs of meningism or headaches (38%). CONCLUSION: The majority of the cases involved meningoradiculitis but other, less common, neurological conditions have been described. The clinical signs suggestive of meningitis are not very marked and might delay the diagnosis.
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Neuroborreliosis de Lyme/complicaciones , Adolescente , Astenia/etiología , Niño , Preescolar , Femenino , Fiebre/etiología , Francia/epidemiología , Cefalea/etiología , Humanos , Neuroborreliosis de Lyme/epidemiología , Masculino , Pediatría/métodos , Pediatría/estadística & datos numéricos , Estudios RetrospectivosRESUMEN
Nijmegen breakage syndrome caused by biallelic pathogenic variants of the DNA-damage response gene NBN, is characterized by severe microcephaly, cancer proneness, infertility, and karyotype abnormalities. We previously reported NBN variants in siblings suffering from fertility defects. Here, we identify a new founder NBN variant (c.442A>G, p.(Thr148Ala)) in Lebanese patients associated with isolated infertility. Functional analyses explored preserved or altered functions correlated with their remarkably mild phenotype. Transcript and protein analyses supported the use of an alternative transcript with in-frame skipping of exons 4-5, leading to p84-NBN protein with a preserved forkhead-associated (FHA) domain. The level of NBN was dramatically reduced and the MRN complex delocalized to the cytoplasm. Interestingly, ataxia-elangiectasia mutated (ATM) also shifted from the nucleus to the cytoplasm, suggesting some interaction between ATM and the MRN complex at a steady state. The ATM pathway activation, attenuated in typical patients with NBS, appeared normal under camptothecin treatment in these new NBN-related infertile patients. Cell cycle checkpoint defect was present in these atypical patients, although to a lesser extent than in typical patients with NBS. In conclusion, we report three new NBN-related infertile patients and we suggest that preserved FHA domain could be responsible for the mild phenotype and intermediate DNA-damage response defects.
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Proteínas de Ciclo Celular/genética , Reparación del ADN , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Variación Genética , Infertilidad/diagnóstico , Infertilidad/genética , Proteínas Nucleares/genética , Adulto , Proteínas de la Ataxia Telangiectasia Mutada/genética , Proteínas de la Ataxia Telangiectasia Mutada/metabolismo , Proteínas de Ciclo Celular/metabolismo , Análisis Mutacional de ADN , Femenino , Citometría de Flujo , Regulación de la Expresión Génica , Estudios de Asociación Genética/métodos , Humanos , Infertilidad/metabolismo , Masculino , Síndrome de Nijmegen/diagnóstico , Síndrome de Nijmegen/genética , Síndrome de Nijmegen/metabolismo , Proteínas Nucleares/metabolismo , Unión Proteica , Transducción de SeñalRESUMEN
BACKGROUND: Short treatments for acute bone and joint infections (BJI) are recommended. We implemented a protocol in 2009 to improve diagnosis and bacteriological documentation, and to shorten antibiotic therapies as per French guidelines (French Pediatric Infectious Disease Group, GPIP). METHODS: To assess the impact of the new clinical protocol for BJI, we conducted a retrospective study from January 1st, 2006 to August 31st, 2012. Two successive cohorts were compared, before and after protocol implementation. All children suspected of community-acquired BJI were included. Confirmed osteomyelitis and septic arthritis required a positive bacterial isolate; otherwise, cases were considered probable. We compared clinical, biological, and radiological data; duration of antibiotic therapy and hospital length of stay; and complications and sequelae. RESULTS: A total of 377 children with suspected BJI were included. The bacteriological identification improved from 32% to 44% when patients were completely evaluated. Isolated bacteria were Staphylococcus aureus (53%), Kingella kingae (17%), Streptococcus pyogenes (15%), and Streptococcus pneumoniae (8%). Before protocol implementation, 70% of patients had a central venous line versus 9% after implementation. Mean duration of IV antibiotics (11 days versus 6 days), mean duration of total antibiotic therapy (45 days versus 32 days) and mean hospital length of stay (13 days versus 7 days) had significantly improved. CONCLUSION: Improvement in bacteriological diagnosis and shorter antibiotic regimens lead to shorter hospital length of stays with no additional morbidity. Simplifying the protocol and better diffusion among health professionals should contribute to shortening BJI treatment duration.
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Antibacterianos/uso terapéutico , Artritis Infecciosa/diagnóstico , Artritis Infecciosa/tratamiento farmacológico , Infecciones Bacterianas/tratamiento farmacológico , Adhesión a Directriz , Osteomielitis/diagnóstico , Osteomielitis/tratamiento farmacológico , Adolescente , Artritis Infecciosa/microbiología , Niño , Preescolar , Femenino , Francia , Humanos , Lactante , Recién Nacido , Masculino , Osteomielitis/microbiología , Guías de Práctica Clínica como Asunto , Estudios RetrospectivosRESUMEN
Ataxia-telangiectasia (A-T) is a recessive disorder caused by biallelic pathogenic variants of ataxia-telangiectasia mutated (ATM). This disease is characterized by progressive ataxia, telangiectasia, immune deficiency, predisposition to malignancies, and radiosensitivity. However, hypomorphic variants may be discovered associated with very atypical phenotypes, raising the importance of evaluating their pathogenic effects. In this study, multiple functional analyses were performed on lymphoblastoid cell lines from 36 patients, comprising 49 ATM variants, 24 being of uncertain significance. Thirteen patients with atypical phenotype and presumably hypomorphic variants were of particular interest to test strength of functional analyses and to highlight discrepancies with typical patients. Western-blot combined with transcript analyses allowed the identification of one missing variant, confirmed suspected splice defects and revealed unsuspected minor transcripts. Subcellular localization analyses confirmed the low level and abnormal cytoplasmic localization of ATM for most A-T cell lines. Interestingly, atypical patients had lower kinase defect and less altered cell-cycle distribution after genotoxic stress than typical patients. In conclusion, this study demonstrated the pathogenic effects of the 49 variants, highlighted the strength of KAP1 phosphorylation test for pathogenicity assessment and allowed the establishment of the Ataxia-TeLangiectasia Atypical Score to predict atypical phenotype. Altogether, we propose strategies for ATM variant detection and classification.
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Proteínas de la Ataxia Telangiectasia Mutada/genética , Ataxia Telangiectasia/diagnóstico , Ataxia Telangiectasia/genética , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Variación Genética , Empalme Alternativo , Ciclo Celular , Línea Celular , Análisis Mutacional de ADN , Estudios de Asociación Genética/métodos , Genotipo , Humanos , Mutación , FenotipoRESUMEN
Ataxia-telangiectasia-like disorder (ATLD) is a rare genomic instability syndrome caused by biallelic variants of MRE11 (meiotic recombination 11) characterized by progressive cerebellar ataxia and typical karyotype abnormalities. These symptoms are common to those of ataxia-telangiectasia, which is consistent with the key role of MRE11 in ataxia-telangiectasia mutated (ATM) activation after DNA double-strand breaks. Three unrelated French patients were referred with ataxia. Only one had typical karyotype abnormalities. Unreported biallelic MRE11 variants were found in these three cases. Interestingly, one variant (c.424G>A) was present in two cases and haplotype analysis strongly suggested a French founder variant. Variants c.544G>A and c.314+4_314+7del lead to splice defects. The level of MRE11 in lymphoblastoid cell lines was consistently and dramatically reduced. Functional consequences were evaluated on activation of the ATM pathway via phosphorylation of ATM targets (KAP1 and CHK2), but no consistent defect was observed. However, an S-phase checkpoint activation defect after camptothecin was observed in these patients with ATLD. In conclusion, we report the first three French ATLD patients and a French founder variant, and propose an S-phase checkpoint activation study to evaluate the pathogenicity of MRE11 variants.
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Ataxia Telangiectasia/diagnóstico , Ataxia Telangiectasia/etiología , Proteínas de la Ataxia Telangiectasia Mutada/genética , Proteínas de la Ataxia Telangiectasia Mutada/metabolismo , Línea Celular Tumoral , Niño , Susceptibilidad a Enfermedades , Femenino , Perfilación de la Expresión Génica , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Haplotipos , Humanos , Lactante , Proteína Homóloga de MRE11/genética , Proteína Homóloga de MRE11/metabolismo , Imagen por Resonancia Magnética , Mutación , Fenotipo , Empalme del ARN , Puntos de Control de la Fase S del Ciclo Celular/genética , Transducción de Señal , TranscriptomaRESUMEN
BACKGROUND: V(D)J recombination ensures the diversity of the adaptive immune system. Although its complete defect causes severe combined immunodeficiency (ie, T-B- severe combined immunodeficiency), its suboptimal activity is associated with a broad spectrum of immune manifestations, such as late-onset combined immunodeficiency and autoimmunity. The earliest molecular diagnosis of these patients is required to adopt the best therapy strategy, particularly when it involves a myeloablative conditioning regimen for hematopoietic stem cell transplantation. OBJECTIVE: We aimed at developing biomarkers based on analysis of the T-cell receptor (TCR) α repertoire to assist in the diagnosis of patients with primary immunodeficiencies with V(D)J recombination and DNA repair deficiencies. METHODS: We used flow cytometric (fluorescence-activated cell sorting) analysis to quantify TCR-Vα7.2-expressing T lymphocytes in peripheral blood and developed PROMIDISα, a multiplex RT-PCR/next-generation sequencing assay, to evaluate a subset of the TCRα repertoire in T lymphocytes. RESULTS: The combined fluorescence-activated cell sorting and PROMIDISα analyses revealed specific signatures in patients with V(D)J recombination-defective primary immunodeficiencies or ataxia telangiectasia/Nijmegen breakage syndromes. CONCLUSION: Analysis of the TCRα repertoire is particularly appropriate in a prospective way to identify patients with partial immune defects caused by suboptimal V(D)J recombination activity, a DNA repair defect, or both. It also constitutes a valuable tool for the retrospective in vivo functional validation of variants identified through exome or panel sequencing. Its broader implementation might be of interest to assist early diagnosis of patients presenting with hypomorphic DNA repair defects inclined to experience acute toxicity during prehematopoietic stem cell transplantation conditioning.
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Síndromes de Inmunodeficiencia , Receptores de Antígenos de Linfocitos T alfa-beta , Recombinación V(D)J/inmunología , Adolescente , Adulto , Niño , Preescolar , Femenino , Humanos , Síndromes de Inmunodeficiencia/genética , Síndromes de Inmunodeficiencia/inmunología , Síndromes de Inmunodeficiencia/patología , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Receptores de Antígenos de Linfocitos T alfa-beta/genética , Receptores de Antígenos de Linfocitos T alfa-beta/inmunología , Estudios RetrospectivosRESUMEN
In modern human cultures where social hierarchies are ubiquitous, people typically signal their hierarchical position through consumption of positional goods-goods that convey one's social position, such as luxury products. Building on animal research and early correlational human studies linking the sex steroid hormone testosterone with hierarchical social interactions, we investigate the influence of testosterone on men's preferences for positional goods. Using a placebo-controlled experiment (N = 243) to measure individuals' desire for status brands and products, we find that administering testosterone increases men's preference for status brands, compared to brands of similar perceived quality but lower perceived status. Furthermore, testosterone increases positive attitudes toward positional goods when they are described as status-enhancing, but not when they are described as power-enhancing or high in quality. Our results provide novel causal evidence for the biological roots of men's preferences for status, bridging decades of animal behavioral studies with contemporary consumer research.
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Comportamiento del Consumidor , Jerarquia Social , Masculinidad , Clase Social , Testosterona/fisiología , Adolescente , Adulto , Humanos , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
Germline DICER1 pathogenic variants predispose to numerous benign and malignant tumors. In this report, we describe DICER1 gene analysis in an adolescent diagnosed with multinodular goiter, ovarian Sertoli-Leydig cell tumor, and lung cyst. DICER1 mutational screening at the DNA level failed to detect any pathogenic variant. Subsequent messenger RNA (mRNA) analysis revealed a 132 nucleotide intronic sequence exonization. This truncating event was caused by a deep intronic mutation generating a de novo acceptor splice site. This study demonstrates that some undetected DICER1 mutations should be investigated at the mRNA level.
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ARN Helicasas DEAD-box/genética , Mutación de Línea Germinal , Bocio Nodular/genética , Intrones/genética , Enfermedades Pulmonares/genética , Neoplasias Ováricas/genética , Ribonucleasa III/genética , Tumor de Células de Sertoli-Leydig/genética , Adolescente , Anciano , Niño , Femenino , Bocio Nodular/patología , Humanos , Enfermedades Pulmonares/patología , Masculino , Neoplasias Ováricas/patología , Linaje , Pronóstico , Tumor de Células de Sertoli-Leydig/patologíaRESUMEN
Whether the recessive ataxias, Ataxia with oculomotor apraxia type 1 (AOA1) and 2 (AOA2) and Ataxia telangiectasia (AT), can be distinguished by video-oculography and alpha-fetoprotein level remains unknown. We compared 40 patients with AOA1, AOA2 and AT, consecutively referred between 2008 and 2015 with 17 healthy subjects. Video-oculography revealed constant impairments in patients such as cerebellar signs, altered fixation, impaired pursuit, hypometric saccades and abnormal antisaccades. Horizontal saccade latencies could be highly increased reflecting oculomotor apraxia in one third of patients. Specific distinctive alpha-fetoprotein thresholds were determined for AOA1 (7-15 µg/L), AOA2 (15-65 µg/L) and AT (>65 µg/L). Early age onset, severe walking disability, movement disorders, sensori-motor neuropathy and cerebellar atrophy were all shared. In conclusion, alpha-fetoprotein level seems to permit a distinction while video-oculography does not and therefore is not mandatory, even if an appropriate oculomotor examination remains crucial. Our findings are that AOA1, AOA2 and AT form a particular group characterized by ataxia with complex oculomotor disturbances and elevated AFP for which the final diagnosis is relying on genetic analysis. These findings could guide genetic analysis, assist reverse-phenotyping and provide background for the interpretation of the numerous variants of unknown significance provided by next-generation sequencing.
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Apraxias/congénito , Ataxia Telangiectasia/sangre , Ataxia Telangiectasia/diagnóstico por imagen , Síndrome de Cogan/sangre , Síndrome de Cogan/diagnóstico por imagen , Imagen Multimodal , alfa-Fetoproteínas/metabolismo , Adolescente , Adulto , Apraxias/sangre , Apraxias/diagnóstico por imagen , Apraxias/genética , Ataxia Telangiectasia/genética , Niño , Preescolar , Síndrome de Cogan/genética , Femenino , Humanos , Masculino , Persona de Mediana Edad , alfa-Fetoproteínas/genéticaRESUMEN
BACKGROUND: The Patient Assessment of Constipation-Symptoms (PAC-SYM) questionnaire is frequently used in clinical trials of constipation. However, the threshold for reduction in total PAC-SYM score used to define a clinical response on this 0-4 point scale has not undergone formal appraisal, and its relationship with clinical benefit as perceived by patients has not been defined. AIM: To determine the minimal important difference in PAC-SYM score, and the optimum cut-off value for defining responders. METHODS: The minimal important difference was estimated using data from six international phase 3/4, double-blind, randomised controlled trials of prucalopride in patients with chronic constipation (NCT01147926, NCT01424228, NCT01116206, NCT00485940, NCT00483886, NCT00488137), with anchor- and distribution-based approaches. Five appropriate patient-reported outcomes were selected as anchors. In addition, receiver operating characteristics (ROC) curve analyses were used to investigate responder discrimination for each anchor. RESULTS: Data from 2884 patients were included. Minimal important difference estimates ranged from -0.52 to -0.63 across the five anchors. Estimates were not affected by study location but were consistently lower for rectal symptoms than for abdominal and stool symptoms. Distribution-based estimates were considerably lower than anchor-based estimates. ROC curve analyses showed optimum cut-off scores for discriminating responders to be similar to anchor-based minimal important difference estimates. CONCLUSIONS: Anchor-based methods gave consistent results for the minimal important difference, at approximately -0.6, and this value was close to the ROC-determined optimal cut-off scores for responder discrimination. This value could be considered in clinical practice. A slightly more conservative threshold (eg -0.75) could be used in clinical trials to reduce the placebo response rate.
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Estreñimiento/fisiopatología , Ensayos Clínicos Controlados Aleatorios como Asunto/métodos , Encuestas y Cuestionarios/normas , Enfermedad Crónica , Estreñimiento/tratamiento farmacológico , Método Doble Ciego , Humanos , Curva ROC , Reproducibilidad de los ResultadosRESUMEN
The aim of this work was to determine the enzymatic activity of phosphofructokinase (PFK), malate dehydrogenase (MDH) and isocitrate dehydrogenase (IDH) in boar spermatozoa and study their participation in bicarbonate-induced capacitation and follicular fluid-induced acrosome reaction. Enzymatic activity of these enzymes was determined spectrophotometrically in extracts of boar spermatozoa. Sperm suspensions were incubated in the presence of bicarbonate (40 mM), a well-known capacitation inducer, or follicular fluid (30%), as an acrosome reaction inducer, and different concentrations of oxoglutarate, oxalomalate and hydroxymalonate, inhibitors of PFK, IDH and MDH, respectively. Capacitation percentages were determined by the fluorescence technique of chlortetracycline (CTC), and true acrosome reaction was determined by trypan blue and differential-interferential contrast, optical microscopy. The activity of PFK in boar spermatozoa enzymatic extracts was 1.70 ± 0.19 U/1010 spermatozoa, the activity of NAD- and NADP-dependent IDH was 0.111 ± 0.005 U/1010 and 2.22 ± 0.14 U/1010 spermatozoa, respectively, and the activity of MDH was 4.24 ± 0.38 U/1010 spermatozoa. The addition of the specific inhibitors of these enzymes prevented sperm capacitation and decreased sperm motility during capacitation and inhibited the acrosome reaction (AR), without affecting the sperm motility during this process. Our results demonstrate the participation of PFK, IDH and MDH in bicarbonate-induced capacitation and follicular fluid-induced acrosome reaction in boar spermatozoa, contributing to elucidate the mechanisms that produce energy necessary for these processes in porcine spermatozoa.
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Reacción Acrosómica/efectos de los fármacos , Isocitrato Deshidrogenasa/metabolismo , Malato Deshidrogenasa/metabolismo , Fosfofructoquinasas/metabolismo , Capacitación Espermática/efectos de los fármacos , Espermatozoides/enzimología , Animales , Bicarbonatos/farmacología , Femenino , Líquido Folicular/fisiología , Isocitrato Deshidrogenasa/antagonistas & inhibidores , Malato Deshidrogenasa/antagonistas & inhibidores , Masculino , Fosfofructoquinasas/antagonistas & inhibidores , Motilidad Espermática/efectos de los fármacos , Espermatozoides/efectos de los fármacos , Sus scrofa , Tartronatos/farmacologíaRESUMEN
Fanconi anemia (FA) is a recessive genetic disease characterized by congenital abnormalities, chromosome instability, progressive bone marrow failure (BMF), and a strong predisposition to cancer. Twenty FA genes have been identified, and the FANC proteins they encode cooperate in a common pathway that regulates DNA crosslink repair and replication fork stability. We identified a child with severe BMF who harbored biallelic inactivating mutations of the translesion DNA synthesis (TLS) gene REV7 (also known as MAD2L2), which encodes the mutant REV7 protein REV7-V85E. Patient-derived cells demonstrated an extended FA phenotype, which included increased chromosome breaks and G2/M accumulation upon exposure to DNA crosslinking agents, γH2AX and 53BP1 foci accumulation, and enhanced p53/p21 activation relative to cells derived from healthy patients. Expression of WT REV7 restored normal cellular and functional phenotypes in the patient's cells, and CRISPR/Cas9 inactivation of REV7 in a non-FA human cell line produced an FA phenotype. Finally, silencing Rev7 in primary hematopoietic cells impaired progenitor function, suggesting that the DNA repair defect underlies the development of BMF in FA. Taken together, our genetic and functional analyses identified REV7 as a previously undescribed FA gene, which we term FANCV.
Asunto(s)
Anemia de Fanconi/genética , Proteínas Mad2/genética , Mutación , Alelos , Animales , Ciclo Celular , Línea Celular Tumoral , Niño , Inestabilidad Cromosómica , Rotura Cromosómica , Estudios de Cohortes , Reactivos de Enlaces Cruzados/química , Daño del ADN , Reparación del ADN , Femenino , Fibroblastos/metabolismo , Silenciador del Gen , Prueba de Complementación Genética , Predisposición Genética a la Enfermedad , Variación Genética , Células Madre Hematopoyéticas/citología , Humanos , Lentivirus , Proteínas Mad2/metabolismo , Masculino , Ratones , Ratones Noqueados , Mitosis , FenotipoRESUMEN
The circadian dynamics of important neuroendocrine-immune mediators have been implicated in progression of rheumatoid arthritis pathophysiology, both clinically as well as in animal models. We present a mathematical model that describes the circadian interactions between mediators of the hypothalamic-pituitary-adrenal (HPA) axis and the proinflammatory cytokines. Model predictions demonstrate that chronically elevated cytokine expression results in the development of adrenal insufficiency and circadian variability in paw edema. Notably, our model also predicts that an increase in mean secretion of corticosterone (CST) after the induction of the disease is accompanied by a decrease in the amplitude of the CST oscillation. Furthermore, alterations in the phase of circadian oscillation of both cytokines and HPA axis mediators are observed. Therefore, by incorporating the circadian interactions between the neuroendocrine-immune mediators, our model is able to simulate important features of rheumatoid arthritis pathophysiology.
Asunto(s)
Artritis Experimental/metabolismo , Artritis Reumatoide/metabolismo , Ritmo Circadiano , Corticosterona/metabolismo , Citocinas/inmunología , Sistema Hipotálamo-Hipofisario/metabolismo , Sistema Hipófiso-Suprarrenal/metabolismo , Receptores de Glucocorticoides/metabolismo , Hormona Adrenocorticotrópica/metabolismo , Animales , Artritis Experimental/inmunología , Artritis Reumatoide/inmunología , Ritmo Circadiano/inmunología , Hormona Liberadora de Corticotropina/metabolismo , Modelos Teóricos , RoedoresRESUMEN
Pseudomonas aeruginosa has been recovered in hospitals from many different sources including sinks and taps. Because P. aeruginosa is one of the main agents of nosocomial infections and increasingly resistant to antibiotics, environmental reservoirs in hospital settings are of great concern. We report here on a cluster of five cases of infection by P. aeruginosa expressing VIM carbapenemases (VIM-PA) in a nephrology intensive care unit. Our investigation pointed to transmission of VIM-PA via hands related to a contaminated tap. VIM-PA may be cross-transmitted to other patients if an environmental reservoir exists. Sinks and taps should be well designed and thoroughly cleaned and disinfected, and use of alcohol hand rub should be promoted.