RESUMO
OBJECTIVE: As exercise intolerance and exercise-induced myalgia are commonly encountered in metabolic myopathies, functional screening tests are commonly used during the diagnostic work-up. Our objective was to evaluate the accuracy of isometric handgrip test (IHT) and progressive cycle ergometer test (PCET) to identify McArdle disease and myoadenylate deaminase (MAD) deficiency and to propose diagnostic algorithms using exercise-induced lactate and ammonia variations. METHODS: A prospective sample of 46 patients underwent an IHT and a PCET as part of their exercise-induced myalgia and intolerance evaluation. The two diagnostics tests were compared against the results of muscle biopsy and/or the presence of mutations in PYGM. A total of 6 patients had McArdle disease, 5 a complete MAD deficiency (MAD absent), 12 a partial MAD deficiency, and 23 patients had normal muscle biopsy and acylcarnitine profile (disease control). RESULTS: The two functional tests could diagnose all McArdle patients with statistical significance, combining a low lactate variation (IHT: <1 mmol/L, AUC = 0.963, P < .0001; PCET: <1 mmol/L, AUC = 0.990, P < .0001) and a large ammonia variation (IHT: >100 µmol/L, AUC = 0.944, P = .0005; PCET: >20 µmol/L, AUC = 1). PCET was superior to IHT for MAD absent diagnosis, combining very low ammonia variation (<10 µmol/L, AUC = 0.910, P < .0001) and moderate lactate variation (>1 mmol/L). CONCLUSIONS: PCET-based decision tree was more accurate than IHT, with respective generalized squared correlations of 0.796 vs 0.668. IHT and PCET are both interesting diagnostic tools to identify McArdle disease, whereas cycle ergometer exercise is more efficient to diagnose complete MAD deficiency.
Assuntos
AMP Desaminase/deficiência , Algoritmos , Teste de Esforço/métodos , Doença de Depósito de Glicogênio Tipo V/diagnóstico , Força da Mão/fisiologia , AMP Desaminase/genética , Adolescente , Adulto , Exercício Físico/fisiologia , Feminino , Doença de Depósito de Glicogênio Tipo V/genética , Doença de Depósito de Glicogênio Tipo V/fisiopatologia , Humanos , Masculino , Pessoa de Meia-Idade , Mutação/genética , Estudos Prospectivos , Adulto JovemRESUMO
In 2014, a botulism outbreak in a flock of laying hens was investigated in France. In the flock of 5020 hens, clinical signs of botulism occurred at 46 weeks of age. A type C/D botulism outbreak was confirmed using the mouse lethality assay for detection of botulinum toxin in serum and a real-time PCR test to detect Clostridium botulinum in intestinal contents. The disease lasted one week with a mortality rate of 2.6% without recurrence. Botulism in laying hens has rarely been reported. Five monthly visits were made to the farm between December 2014 and May 2015 for a longitudinal study of the persistence of C. botulinum in the poultry house after the outbreak, and to assess egg contamination by C. botulinum. Several samples were collected on each visit: in the house (from the ventilation circuit, the egg circuit, water and feed, droppings) and the surrounding area. Thirty clean and 30 dirty eggs were also swabbed at each visit. In addition, 12 dirty and 12 clean eggs were collected to analyse eggshell and egg content. The samples were analysed using real-time PCR to detect type C/D C. botulinum. The bacterium was still detected in the house more than 5 months after the outbreak, mostly on the walls and in the egg circuit. Regarding egg contamination, the bacteria were detected only on the shell but not in the content of the eggs. Control measures should therefore be implemented throughout the egg production period to avoid dissemination of the bacteria, particularly during egg collection.
Assuntos
Toxinas Botulínicas/sangue , Botulismo/veterinária , Galinhas/microbiologia , Clostridium botulinum/isolamento & purificação , Surtos de Doenças/veterinária , Doenças das Aves Domésticas/microbiologia , Animais , Botulismo/epidemiologia , Botulismo/microbiologia , Clostridium botulinum/genética , Ovos/microbiologia , Feminino , França/epidemiologia , Estudos Longitudinais , Camundongos , Doenças das Aves Domésticas/epidemiologia , Reação em Cadeia da Polimerase em Tempo Real/veterináriaRESUMO
BACKGROUND: The operating theatre, anaesthesia induction and separation from parents create fear and anxiety in children. Anxiety leads to adverse behavioral changes appearing and sometimes persisting during the postoperative period. Our aim was to compare the effects of midazolam (0.3 mg kg-1: MDZ) for premedication with age-appropriate tablet game apps (TAB) on children anxiety during and after ambulatory surgery. METHODS: A randomized controlled trial was conducted from May 16th, 2013 to March 25th, 2014 at the Children Hospital of Lyon. The primary outcome of this study was the change in m-YPAS score at the time of anaesthetic mask induction. Anxiety was also assessed in the waiting surgical area, at the time of separation with parents and when back in the ambulatory surgery ward. RESULTS: One hundred and eighteen patients aged four-11 yr were recruited, 60 in the TAB Group and 58 in the MDZ Group. Main endpoint was missing for three patients from the MDZ Group. At the time of mask induction, there was no significant difference between MDZ and TAB Group for the m-YPAS score (40.5 (18.6) vs 41.8 (20.7), P = 0.99). There was no significant correlation between m-YPAS score and its evolution over the four period of time between subjects. CONCLUSIONS: We were not able to show whether TAB is superior to MDZ to blunt anxiety in children undergoing ambulatory surgery. TAB is a non-pharmacological tool which has the capacity in reducing perioperative stress without any sedative effect in this population. CLINICAL TRIAL REGISTRATION: NCT 02192710.
Assuntos
Procedimentos Cirúrgicos Ambulatórios , Ansiolíticos/uso terapêutico , Midazolam/uso terapêutico , Pais/psicologia , Medicação Pré-Anestésica , Jogos de Vídeo , Criança , Pré-Escolar , Computadores de Mão , Feminino , Humanos , MasculinoRESUMO
BACKGROUND: Patients suffering from haemoglobinopathies may be treated by red blood cell (RBC) transfusion on a regular basis and then exposed to multiple antigens with a recurrent, potential risk of alloimmunization routinely prevented by extended RBC antigen cross-matching. While time-consuming and labour-intensive serological analyses are the gold standard for RBC typing, genotyping by current high-throughput molecular tools, including next-generation sequencing (NGS), appears to offer a potent alternative. STUDY DESIGN AND METHODS: The potential of extended blood group genotyping (EBGG) by NGS of 17 genes involved in 14 blood group systems was evaluated in a cohort of 48 patients with sickle-cell disease. Sample preparation and sequencing were simplified and automated for future routine implementation. RESULTS: Sequencing data were obtained for all DNA samples with two different sequencing machines. Prediction of phenotypes could be made in 12 blood group systems and partially in two other blood group systems (Rh and MNS). Importantly, predicted phenotypes in the MNS (S/s), Duffy, Kidd and Kell systems matched well with serological data (98·9%), when available. Unreferenced alleles in the ACHE and ART4 genes, respectively, involved in the Yt and Dombrock blood groups, were identified, then contributing to extend the current knowledge of blood group molecular genetics. CONCLUSIONS: Overall, we consider that our strategy for NGS-based EBGG, assisted by a simple method for genotyping exons 1 and 2 of the pairs of homologous genes (i.e. RHD/RHCE and GYPA/GYPB), as well as the future support of potent bioinformatics tools, may be implemented for routine diagnosis in specific populations.
Assuntos
Anemia Falciforme/genética , Anemia Falciforme/terapia , Sequência de Bases , Tipagem e Reações Cruzadas Sanguíneas , Segurança do Sangue , Análise Mutacional de DNA , Transfusão de Eritrócitos , Frequência do Gene , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Sistema do Grupo Sanguíneo de Kell/genética , FenótipoRESUMO
Between 2011 and 2013, 17 poultry botulism outbreaks were investigated in France. All cases were associated with Clostridium botulinum type C-D. Presence of C. botulinum was studied in seven areas: poultry house, changing room, ventilation system, surroundings, animal reservoirs, water, and feed. Swabs, litter, soil, darkling beetles, rodents and wild bird droppings, feed and water samples were collected. The presence of C. botulinum type C-D in the environment of affected flocks was detected in 39.5% of the 185 samples analysed by real-time polymerase chain reaction. C. botulinum type C-D was reported in each area. Four areas were more frequently contaminated, being found positive in more than one-half of farms: darkling beetles (9/11), poultry house (14/17), water (13/16) and surroundings (11/16). After cleaning and disinfection, the ventilation system and/or the soil (in the houses and the surroundings) returned positive results in four out of eight poultry farms. Consequently, darkling beetles, the drinking water, the ventilation system and the soil in the surroundings and the houses were identified as the main critical contaminated areas to consider in poultry farms to prevent recurrence of botulism outbreaks.
Assuntos
Botulismo/veterinária , Clostridium botulinum/isolamento & purificação , Abrigo para Animais/normas , Doenças das Aves Domésticas/microbiologia , Aves Domésticas , Animais , Botulismo/epidemiologia , Botulismo/microbiologia , Surtos de Doenças/veterinária , Microbiologia Ambiental , Feminino , França/epidemiologia , Masculino , Doenças das Aves Domésticas/epidemiologia , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Decompression sickness (DCS) is a systemic syndrome that can occur after an environmental pressure reduction. Previously, we showed that the plasmatic tetrameric form of transthyretin (TTR) nearly disappeared in rats suffering DCS but not in asymptomatic ones. In this pilot study, we assessed whether the resistance to DCS could be associated with polymorphism of the gene of TTR. For this study, Sanger sequencing was performed on purified PCR products from the liver of 14-week-old male and female standard and DCS-resistant rats (n = 5 per group). Hepatic TTR mRNA expression was assessed by RT-qPCR in 18-19 week-old male and female standard and resistant rats (n = 6 per group). There is a synonymous single nucleotide polymorphism (SNP) on the third base of codon 46 (c.138 C > T). The thymine allele was present in 90% and 100% of males and females standard, respectively. However, this allele is present in only 30% of DCS-resistant males and females (p = 0.0002301). In the liver, there is a significant effect of the resistance to DCS (p = 0.043) and sex (p = 0.047) on TTR expression. Levels of TTR mRNA were lower in DCS-resistant animals. To conclude, DCS resistance might be associated with a SNP and a lower expression of TTR.
Assuntos
Doença da Descompressão , Fígado , Polimorfismo de Nucleotídeo Único , Pré-Albumina , Animais , Masculino , Pré-Albumina/genética , Pré-Albumina/metabolismo , Feminino , Projetos Piloto , Ratos , Doença da Descompressão/genética , Doença da Descompressão/metabolismo , Fígado/metabolismo , Resistência à Doença/genética , Ratos Wistar , RNA Mensageiro/metabolismo , RNA Mensageiro/genéticaRESUMO
1. The growth of Bacillus cereus group bacteria often limits the shelf-life of pasteurised liquid egg products and is also a putative toxin producer. This study was performed to better understand the route of contamination by B. cereus in egg products by studying the factors affecting eggshell contamination on-farm. 2. Eggs were collected in warm and cold seasons in 50 conventional laying farms in Western France. Egg surfaces were analysed for the presence of B. cereus group bacteria, environmental measurements were recorded and production practices were identified through a questionnaire filled out by the farmers. 3. A total of 44% of the farms were contaminated by mesophilic and 10% by psychrotrophic B. cereus group bacteria. No significant effect of the season was observed, whatever the thermal type. Several procedures were associated with reduced eggshell contamination by mesophilic bacteria, including the limitation of dust formation from manure and feeding and efficient disinfection of the silo, houses and the sanitary wall between houses. 4. The research highlights the need to promote prevention strategies, from farm to fork, for the control of eggshell and putative subsequent egg product contamination by B. cereus group bacteria.
Assuntos
Bacillus cereus/isolamento & purificação , Galinhas/microbiologia , Casca de Ovo/microbiologia , Criação de Animais Domésticos/métodos , Animais , Desinfecção/métodos , Feminino , Microbiologia de Alimentos , França , Abrigo para Animais , Estações do AnoRESUMO
BACKGROUND AND AIMS: Falls among older people remain a major public health challenge. Body-worn sensors are needed to improve the understanding of the underlying mechanisms and kinematics of falls. The aim of this systematic review is to assemble, extract and critically discuss the information available in published studies, as well as the characteristics of these investigations (fall documentation and technical characteristics). METHODS: The searching of publically accessible electronic literature databases for articles on fall detection with body-worn sensors identified a collection of 96 records (33 journal articles, 60 conference proceedings and 3 project reports) published between 1998 and 2012. These publications were analysed by two independent expert reviewers. Information was extracted into a custom-built data form and processed using SPSS (SPSS Inc., Chicago, IL, USA). RESULTS: The main findings were the lack of agreement between the methodology and documentation protocols (study, fall reporting and technical characteristics) used in the studies, as well as a substantial lack of real-world fall recordings. A methodological pitfall identified in most articles was the lack of an established fall definition. The types of sensors and their technical specifications varied considerably between studies. CONCLUSION: Limited methodological agreement between sensor-based fall detection studies using body-worn sensors was identified. Published evidence-based support for commercially available fall detection devices is still lacking. A worldwide research group consensus is needed to address fundamental issues such as incident verification, the establishment of guidelines for fall reporting and the development of a common fall definition.
Assuntos
Acidentes por Quedas/prevenção & controle , Acidentes por Quedas/estatística & dados numéricos , Actigrafia/métodos , Monitorização Ambulatorial/instrumentação , Monitorização Ambulatorial/métodos , Telemedicina/métodos , Actigrafia/instrumentação , Actigrafia/estatística & dados numéricos , Medicina Baseada em Evidências , Humanos , Monitorização Ambulatorial/estatística & dados numéricos , Telemedicina/estatística & dados numéricos , TransdutoresRESUMO
The aim of the present study was (i) to type, by genotypic and phenotypic methods, a collection of psychrotrophic bacteria belonging to the Bacillus cereus group collected in a farm and in 6 egg breaking industries during a period covering a warm and a cold season, and (ii) to characterize the egg product spoilage (growth in liquid whole egg) and the sanitary risk potential (cytotoxic activity on Caco-2 cells and adhesion on stainless steel) of each isolate of the collection. The investigation of specific psychrotrophic and mesophilic signatures together with the study of ability to grow at 6 °C and/or at 43 °C on optimal agar medium allowed highlighting twelve profiles, the major one corresponding to the species Bacillus weihenstephanensis (46.2% of the collection). The diversity of the profiles depended on the season and on the origin of the isolates. In terms of food spoilage, all the isolates were able to grow at the same level in liquid whole egg and in optimal medium, even at low temperature. Under the same conditions, the cytotoxic activity depended on the isolate, the medium and the temperature. At 10 °C, no isolate was cytotoxic at 10 °C in liquid whole egg and only one, belonging to the Bacillus weihenstephansensis species, in the optimal medium. All the isolates were able to adhere on stainless steel at various levels, from 2.6±0.2 log cfu/cm(2) to 4.9±0.1 log cfu/cm(2). A large majority (80.8%) was strongly adhering and could lead to the formation of biofilms in industrial equipments.
Assuntos
Bacillus cereus , Aderência Bacteriana/fisiologia , Qualidade de Produtos para o Consumidor , Ovos/microbiologia , Contaminação de Alimentos/análise , Bacillus cereus/classificação , Bacillus cereus/isolamento & purificação , Bacillus cereus/fisiologia , Biodiversidade , Microbiologia de Alimentos , Humanos , Filogenia , Medição de Risco , Estações do Ano , Aço InoxidávelRESUMO
INTRODUCTION: In the first wave, thrombotic complications were common in COVID-19 patients. It is unknown whether state-of-the-art treatment has resulted in less thrombotic complications in the second wave. METHODS: We assessed the incidence of thrombotic complications and overall mortality in COVID-19 patients admitted to eight Dutch hospitals between September 1st and November 30th 2020. Follow-up ended at discharge, transfer to another hospital, when they died, or on November 30th 2020, whichever came first. Cumulative incidences were estimated, adjusted for competing risk of death. These were compared to those observed in 579 patients admitted in the first wave, between February 24th and April 26th 2020, by means of Cox regression techniques adjusted for age, sex and weight. RESULTS: In total 947 patients with COVID-19 were included in this analysis, of whom 358 patients were admitted to the ICU; 144 patients died (15%). The adjusted cumulative incidence of all thrombotic complications after 10, 20 and 30 days was 12% (95% confidence interval (CI) 9.8-15%), 16% (13-19%) and 21% (17-25%), respectively. Patient characteristics between the first and second wave were comparable. The adjusted hazard ratio (HR) for overall mortality in the second wave versus the first wave was 0.53 (95%CI 0.41-0.70). The adjusted HR for any thrombotic complication in the second versus the first wave was 0.89 (95%CI 0.65-1.2). CONCLUSIONS: Mortality was reduced by 47% in the second wave, but the thrombotic complication rate remained high, and comparable to the first wave. Careful attention to provision of adequate thromboprophylaxis is invariably warranted.
Assuntos
COVID-19/complicações , Embolia Pulmonar/etiologia , Trombose/etiologia , Tromboembolia Venosa/etiologia , Idoso , Idoso de 80 Anos ou mais , COVID-19/mortalidade , Estudos de Coortes , Estado Terminal/mortalidade , Feminino , Hospitalização , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Países Baixos/epidemiologia , Modelos de Riscos Proporcionais , SARS-CoV-2/isolamento & purificaçãoRESUMO
Eleven affected members of a large German-American family segregating recessively inherited, congenital, non-syndromic sensorineural hearing loss (SNHL) were found to be homozygous for the common 35delG mutation of GJB2, the gene encoding the gap junction protein Connexin 26. Surprisingly, four additional family members with bilateral profound SNHL carried only a single 35delG mutation. Previously, we demonstrated reduced expression of both GJB2 and GJB6 mRNA from the allele carried in trans with that bearing the 35delG mutation in these four persons. Using array comparative genome hybridization (array CGH), we have now identified on this allele a deletion of 131.4 kb whose proximal breakpoint lies more than 100 kb upstream of the transcriptional start sites of GJB2 and GJB6. This deletion, del(chr13:19,837,344-19,968,698), segregates as a completely penetrant DFNB1 allele in this family. It is not present in 528 persons with SNHL and monoallelic mutation of GJB2 or GJB6, and we have not identified any other candidate pathogenic copy number variation by arrayCGH in a subset of 10 such persons. Characterization of distant GJB2/GJB6 cis-regulatory regions evidenced by this allele may be required to find the 'missing' DFNB1 mutations that are believed to exist.
Assuntos
Conexinas/genética , Regulação da Expressão Gênica , Sequências Reguladoras de Ácido Nucleico/genética , Deleção de Sequência , Alelos , Sequência de Bases , Deleção Cromossômica , Cromossomos Humanos Par 13/genética , Hibridização Genômica Comparativa , Conexina 26 , Conexina 30 , Saúde da Família , Feminino , Testes Genéticos , Genótipo , Perda Auditiva Neurossensorial/diagnóstico , Perda Auditiva Neurossensorial/genética , Humanos , Masculino , Dados de Sequência Molecular , Linhagem , Penetrância , Homologia de Sequência do Ácido NucleicoRESUMO
BACKGROUND AND AIMS: The prevalence and natural history of hereditary pancreatitis (HP) remain poorly documented. The aims of this study were to assess genetic, epidemiological, clinical and morphological characteristics of HP in an extensive national survey. METHODS: A cohort comprising all HP patients was constituted by contacting all gastroenterologists and paediatricians (response rate 84%) and genetics laboratories (response rate 100%) in France (60,200,000 inhabitants). Inclusion criteria were the presence of mutation in the cationic trypsingen gene (PRSS1 gene), or chronic pancreatitis in at least two first-degree relatives, or three second-degree relatives, in the absence of precipitating factors for pancreatitis. RESULTS: 78 families and 200 patients were included (181 alive, 6673 person-years, males 53%, alcoholism 5%, smoking 34%). The prevalence was 0.3/100,000 inhabitants. PRSS1 mutations were detected in 68% (R122H 78%, N29I 12%, others 10%). Penetrance was 93%. Median age at first symptom, diagnosis and date of last news, were 10 (range 1-73), 19 (1-80) and 30 (1-84) years, respectively. HP was responsible for pancreatic pain (83%), acute pancreatitis (69%), pseudocysts (23%), cholestasis (3%), pancreatic calcifications (61%), exocrine pancreatic insufficiency (34%, median age of occurrence 29 years), diabetes mellitus (26%, median age of occurrence 38 years) and pancreatic adenocarcinoma (5%, median age 55 years). No differences in clinical and morphological data according to genetic status were observed. 19 patients died, including 10 directly from HP (8 from pancreatic adenocarcinoma). CONCLUSION: The prevalence of HP in France is at least 0.3/100,000. PRSS1 gene mutations are found in 2/3 with a 93% penetrance. Mutation type is not correlated with clinical/morphological expression. Pancreatic adenocarcinoma is the cause of nearly half the deaths.
Assuntos
Pancreatite Crônica/genética , Adenocarcinoma/epidemiologia , Adenocarcinoma/etiologia , Adolescente , Adulto , Distribuição por Idade , Idade de Início , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Métodos Epidemiológicos , Insuficiência Pancreática Exócrina/epidemiologia , Insuficiência Pancreática Exócrina/etiologia , Feminino , França/epidemiologia , Genótipo , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Mutação , Neoplasias Pancreáticas/epidemiologia , Neoplasias Pancreáticas/etiologia , Pancreatite Crônica/complicações , Pancreatite Crônica/epidemiologia , Penetrância , Fenótipo , Tripsina , Tripsinogênio/genética , Adulto JovemRESUMO
Staphylococcus aureus is an important pathogen in domestic ruminants. The main objective of this study was to determine the similarity of epidemiologically unrelated S. aureus isolates from bovine, ovine, and caprine hosts regardless the locus of isolation (nares and udder). By pulsed-field gel electrophoresis, seven major pulsotypes were identified among 153 isolates recovered from 12 different regions of France as well as from Brazil, the USA and Belgium. Typing of the accessory gene regulator (agr) and capsular (cap) serotype was carried out on all the isolates and revealed the predominance of agr I and III and of cap8 regardless the ruminant host species. Screening for methicilin-resistant S. aureus (MRSA) was carried out by disk diffusion and revealed a prevalence of only 3.2% of MRSA among the strains tested. These results suggest the existence of a host rather than tissue specificity among S. aureus isolates colonising the ruminant species and suggest a limited transmission of those isolates between large (bovine) and small (ovine-caprine) ruminants. The agr class and cap types correlated with pulsotype clusters rather than with a specific host species. Antimicrobial resistance appears not to have contributed to the predominance of any given genotypes, and MRSA prevalence appears very low in ruminant isolates.
Assuntos
Doenças dos Bovinos/microbiologia , Doenças das Cabras/microbiologia , Doenças dos Ovinos/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus aureus/classificação , Animais , Cápsulas Bacterianas/metabolismo , Bovinos , Regulação Bacteriana da Expressão Gênica , Genótipo , Cabras , Interações Hospedeiro-Patógeno , Filogenia , Polissacarídeos Bacterianos/genética , Polissacarídeos Bacterianos/metabolismo , Ovinos , Infecções Estafilocócicas/microbiologiaRESUMO
The present paper aims to assess the risk of rib fractures caused by any rigid less-lethal kinetic energy projectiles. To that end, a coupled experimental and numerical approach is proposed to relate ballistic experiments with the risk of blunt trauma. A polymer gel block is employed as ballistic testing medium to interpret ballistic impacts through the measurement of the dynamic gel wall displacement. Moreover, a biofidelic 50th percentile human torso finite element model created in the code Hypermesh (Altair HyperWorks ©) is used to replicate experiments and real world accidents. Then, the probability curve of rib fractures is a function of the viscous criterion and derives from a study on human cadaver mid-sternum available in the literature. Twelve impact conditions of rigid projectiles are applied to a SEBS gel block and are replicated on the human torso model mid-sternum. A statistical analysis is performed by virtue of a Spearman's correlation matrix in order to identify relations between experimental measurements and the viscous criterion evaluated numerically. The determination of both statistical significances and correlation coefficients results in several strong correlations between experimental measurements and the viscous criterion evaluated numerically. These relations imply the establishment of transfer functions between experimental metrics on the gel block (the maximum gel wall displacement and a gel wall displacement - rate of displacement based metric) and the probability of rib fractures. Finally, these correlations constitute a primary and an up-and-coming predicting tool for the risk of rib fractures.
Assuntos
Balística Forense , Cinética , Modelos Biológicos , Fraturas das Costelas , Ferimentos não Penetrantes , Análise de Elementos Finitos , Géis , Humanos , Modelos Logísticos , PolímerosRESUMO
In 1996, shortly after a locus for hereditary pancreatitis had been mapped to chromosome 7q35, an apparent gain-of-function missense mutation, p.R122H, in the cationic trypsinogen gene (PRSS1) was identified. Thereafter, the search for chronic pancreatitis-associated genetic factors has been largely focused on one form of genetic variation, namely, single nucleotide substitutions (SNSs). Only very recently has another type of genetic variation - copy number variations (CNVs) - been found to cause the disease. First, we identified duplication and triplication of an approximately 605 kb segment on chromosome 7q35 in French white patients with hereditary or idiopathic chronic pancreatitis. These alterations increased the copy number of PRSS1 as well as PRSS2, which encodes anionic trypsinogen. Second, we characterized a hybrid trypsinogen gene, in which exons 1 and 2 were derived from PRSS2 and exons 3 to 5 from PRSS1. Interestingly, this hybrid gene had two independent gain-of-function effects: increased trypsinogen gene copy number and it contained the p.N29I pancreatitis-causing missense mutation. Lastly, we identified two loss-of-function copy number mutations (deletions) in the SPINK1 gene, which encodes pancreatic secretory trypsin inhibitor (PSTI). Particularly, in one family with chronic pancreatitis, deletion of the complete SPINK1 gene was co-inherited with a CFTR missense mutation (p.L997F), revealing another layer of complexity between CNV and SNS interactions in the determination of a given disease phenotype. These findings represent a further demonstration of how studies of CNVs have altered the landscape of genetic research in the past few years and offer a fresh glimpse into the exciting realm of human CNVs.
Assuntos
Dosagem de Genes/genética , Pancreatite Crônica/genética , Duplicação Gênica , Genoma/genética , Humanos , Mutação/genética , Pancreatite Crônica/classificação , Tripsinogênio/genéticaRESUMO
AIMS: To report on a family with five members who carry the A3243G mutation in mitochondrial tRNA for leucine 1 (MTTL1) and present with diabetes, chronic intestinal pseudo-obstruction (CIPO) and recurrent pancreatitis, and to screen for this mutation in a cohort of 36 unrelated patients with recurrent pancreatitis. METHODS: The mutation was quantified in several tissue samples from patients. Respiratory chain activity was studied in muscle biopsies and fibroblast cultures. In addition, the thymidine phosphorylase gene (TP) involved in mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) and three genes involved in chronic pancreatitis - PRSS1, SPINK1 and CFTR - were sequenced in affected patients. Finally, the MTTL1 gene was examined in 36 unrelated patients who had recurrent pancreatitis, but no mutations in the PRSS1 and SPINK1 genes. RESULTS: Heteroplasmy for the mtDNA A3243G mutation was found in all tissue samples from these patients, but no mutations were found in the genes coding for thymidine phosphorylase, PRSS1, SPINK1 and CFTR. Also, none of the 36 unrelated patients with recurrent pancreatitis were carrying any MTTL1 mutations. CONCLUSION: The mtDNA A3243G mutation associated with the gastrointestinal manifestations observed in the affected family should be regarded as a possible cause of CIPO and unexplained recurrent pancreatitis. However, the mutation is probably only weakly involved in cases of isolated recurrent pancreatitis.
Assuntos
DNA Mitocondrial/genética , Complicações do Diabetes/genética , Diabetes Mellitus/genética , Pseudo-Obstrução Intestinal/genética , Pancreatite/genética , Polimorfismo de Nucleotídeo Único , Surdez/genética , Complicações do Diabetes/patologia , Diabetes Mellitus/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Músculo Esquelético/patologia , Mutação , Linhagem , RecidivaRESUMO
In October 2014, an outbreak of botulism type D/C occurred on two cattle farms in close proximity. A poultry farm located nearby with no history of botulism had transferred poultry manure to both bovine farms before the beginning of the outbreak. Given this context, epidemiological investigation was conducted to determine if the poultry farm was a reservoir of C. botulinum type D/C and to identify the source of contamination on the cattle farms. Environmental samples were collected at three houses on the poultry farm (boot swabs from the surroundings, swabs from the ventilation system, boot swabs from the poultry litter and darkling beetles samples), and on the two cattle farms (silage samples, boot swabs from the cattle stalls, boot swabs from the cattle pasture and poultry manure samples). These samples were analyzed using real-time PCR after an enrichment step to detect C. botulinum type D/C. On the poultry farm, three boot swabs from the surroundings, two swabs from the ventilation system, one boot swab from the litter and one sample of darkling beetles were detected positive. On one cattle farm, C. botulinum type D/C was identified in a sample of silage made from grass grown on a field on which the poultry manure had previously been stored and in a boot swab from a pasture. On the other cattle farm, C. botulinum type D/C was detected in a sample of poultry manure stored on the cattle farm and in a boot swab from a pasture. This investigation shows that the healthy poultry farm might have been the reservoir of C. botulinum type D/C and that cross-contamination between poultry and cattle likely occurred, resulting in the botulism outbreak on the two cattle farms.