RESUMO
BACKGROUND AND AIMS: Around 750,000 patients per year will be cured of HCV infection until 2030. Those with compensated advanced chronic liver disease remain at risk for hepatic decompensation and de novo HCC. Algorithms have been developed to stratify risk early after cure; however, data on long-term outcomes and the prognostic utility of these risk stratification algorithms at later time points are lacking. APPROACH AND RESULTS: We retrospectively analyzed a cohort of 2335 patients with compensated advanced chronic liver disease (liver stiffness measurement≥10 kPa) who achieved HCV-cure by interferon-free therapies from 15 European centers (median age 60.2±11.9 y, 21.1% obesity, 21.2% diabetes).During a median follow-up of 6 years, first hepatic decompensation occurred in 84 patients (3.6%, incidence rate: 0.74%/y, cumulative incidence at 6 y: 3.2%); 183 (7.8%) patients developed de novo HCC (incidence rate: 1.60%/y, cumulative incidence at 6 y: 8.3%), with both risks being strictly linear over time.Baveno VII criteria to exclude (FU-liver stiffness measurement <12 kPa and follow-up platelet count >150 g/L) or rule-in (FU-liver stiffness measurement ≥25 kPa) clinically significant portal hypertension (CSPH) stratified the risk of hepatic decompensation with proportional hazards. Estimated probability of CSPH discriminated patients developing versus not developing hepatic decompensation in the gray zone (ie, patients meeting none of the above criteria).Published HCC risk stratification algorithms identified high-incidence and low-incidence groups; however, the size of the latter group varied substantially (9.9%-69.1%). A granular "HCC-sustained virologic response" model was developed to inform an individual patient's HCC risk after HCV-cure. CONCLUSIONS: In patients with compensated advanced chronic liver disease, the risks of hepatic decompensation and HCC remain constant after HCV-cure, even in the long term (>3 y). One-time post-treatment risk stratification based on noninvasive criteria provides important prognostic information that is maintained during long-term follow-up, as the hazards remain proportional over time.
RESUMO
BACKGROUND & AIMS: Baveno VII has defined a clinically significant (i.e., prognostically meaningful) decrease in liver stiffness measurement (LSM) in cACLD as a decrease of ≥20% associated with a final LSM <20 kPa or any decrease to <10 kPa. However, these rules have not yet been validated against direct clinical endpoints. METHODS: We retrospectively analysed patients with cACLD (LSM ≥10 kPa) with paired liver stiffness measurement (LSM) before (BL) and after (FU) HCV cure by interferon-free therapies from 15 European centres. The cumulative incidence of hepatic decompensation was compared according to these criteria, considering hepatocellular carcinoma and non-liver-related death as competing risks. RESULTS: A total of 2,335 patients followed for a median of 6 years were analysed. Median BL-LSM was 16.6 kPa with 37.1% having ≥20 kPa. After HCV cure, FU-LSM decreased to a median of 10.9 kPa (<10 kPa: 1,002 [42.9%], ≥20 kPa: 465 [19.9%]) translating into a median LSM change of -5.3 (-8.8 to -2.4) kPa corresponding to -33.9 (-48.0 to -15.9) %. Patients achieving a clinically significant decrease (65.4%) had a significantly lower risk of hepatic decompensation (subdistribution hazard ratio: 0.12, 95% CI 0.04-0.35, p <0.001). However, these risk differences were primarily driven by a negligible risk in patients with FU-LSM <10 kPa (5-year cumulative incidence: 0.3%) compared to a high risk in patients with FU-LSM ≥20 kPa (16.6%). Patients with FU-LSM 10-19.9 kPa (37.4%) also had a low risk of hepatic decompensation (5-year cumulative incidence: 1.7%), and importantly, the risk of hepatic decompensation did not differ between those with/without an LSM decrease of ≥20% (p = 0.550). CONCLUSIONS: FU-LSM is key for risk stratification after HCV cure and should guide clinical decision making. LSM dynamics do not hold significant prognostic information in patients with FU-LSM 10-19.9 kPa, and thus, their consideration is not of sufficient incremental value in the specific context of HCV cure. IMPACT AND IMPLICATIONS: Liver stiffness measurement (LSM) is increasingly applied as a prognostic biomarker and commonly decreases in patients with compensated advanced chronic liver disease achieving HCV cure. Although Baveno VII proposed criteria for a clinically significant decrease, little is known about the prognostic utility of LSM dynamics (changes through antiviral therapy). Interestingly, in those with a post-treatment LSM of 10-19.9 kPa, LSM dynamics did not provide incremental information, arguing against the consideration of LSM dynamics as prognostic criteria. Thus, post-treatment LSM should guide the management of patients with compensated advanced chronic liver disease achieving HCV cure.
Assuntos
Técnicas de Imagem por Elasticidade , Hepatite C Crônica , Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Estudos Retrospectivos , Técnicas de Imagem por Elasticidade/métodos , Hepatite C Crônica/tratamento farmacológico , Hepatite C Crônica/complicações , Antivirais/uso terapêutico , Cirrose Hepática/epidemiologia , Prognóstico , Idoso , Fígado/diagnóstico por imagem , Neoplasias Hepáticas/epidemiologia , Neoplasias Hepáticas/etiologia , Adulto , Carcinoma Hepatocelular/epidemiologia , Carcinoma Hepatocelular/etiologiaRESUMO
A new bioinformatic platform (APTERION) was used to design in a short time and with high specificity an aptamer for the detection of the spike protein, a structural protein of SARS-CoV-2 virus, responsible for the COVID-19 pandemic. The aptamer concentration on the carbon electrode surface was optimized using static contact angle and fluorescence method, while specificity was tested using differential pulse voltammetry (DPV) associated to carbon screen-printed electrodes. The data obtained demonstrated the good features of the aptamer which could be used to create a rapid method for the detection of SARS-CoV-2 virus. In fact, it is specific for spike also when tested against bovine serum albumin and lysozyme, competitor proteins if saliva is used as sample to test for the virus presence. Spectrofluorometric characterization allowed to measure the amount of aptamer present on the carbon electrode surface, while DPV measurements proved the affinity of the aptamer towards the spike protein and gave quantitative results. The acquired data allowed to conclude that the APTERION bioinformatic platform is a good method for aptamer design for rapidity and specificity. KEY POINTS: ⢠Spike protein detection using an electrochemical biosensor ⢠Aptamer characterization by contact angle and fluorescent measurements on electrode surface ⢠Computational design of specific aptamers to speed up the aptameric sequence time.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , COVID-19 , Humanos , SARS-CoV-2 , Técnicas Eletroquímicas/métodos , Glicoproteína da Espícula de Coronavírus , Aptâmeros de Nucleotídeos/química , Pandemias , Carbono , Glicoproteínas , Técnicas Biossensoriais/métodos , EletrodosRESUMO
BACKGROUND: Antimicrobial resistance (AMR) is a critical global issue that poses significant threats to human health, animal welfare, and the environment. With the increasing emergence of resistant microorganisms, the effectiveness of current antimicrobial medicines against common infections is diminishing. This study aims to conduct a competitive meta-analysis of surveillance data on resistant microorganisms and their antimicrobial resistance patterns in two countries, Egypt and the United Kingdom (UK). METHODS: Data for this study were obtained from published reports spanning the period from 2013 to 2022. In Egypt and the UK, a total of 9,751 and 10,602 food samples were analyzed, respectively. Among these samples, 3,205 (32.87%) in Egypt and 4,447 (41.94%) in the UK were found to contain AMR bacteria. RESULTS: In Egypt, the predominant resistance was observed against ß-lactam and aminoglycosides, while in the United Kingdom, most isolates exhibited resistance to tetracycline and ß-lactam. The findings from the analysis underscore the increasing prevalence of AMR in certain microorganisms, raising concerns about the development of multidrug resistance. CONCLUSION: This meta-analysis sheds light on the escalating AMR problem associated with certain microorganisms that pose a higher risk of multidrug resistance development. The significance of implementing One Health AMR surveillance is emphasized to bridge knowledge gaps and facilitate accurate AMR risk assessments, ensuring consumer safety. Urgent actions are needed on a global scale to combat AMR and preserve the effectiveness of antimicrobial treatments for the well-being of all living beings.
Assuntos
Anti-Infecciosos , Saúde Única , Animais , Humanos , Antibacterianos/uso terapêutico , beta-Lactamas , Farmacorresistência Bacteriana , Egito , Reino UnidoRESUMO
Foods contaminated by pathogens are responsible for foodborne diseases which have socioeconomic impacts. Many approaches have been extensively investigated to obtain specific and sensitive methods to detect pathogens in food, but they are often not easy to perform and require trained personnel. This work aims to propose a textile organic electrochemical transistor-based (OECT) biosensor to detect L. monocytogenes in food samples. The analyses were performed with culture-based methods, Listeria Precis™ method, PCR, and our textile OECT biosensor which used poly(3,4-ethylenedioxythiophene) (PEDOT):polystyrene sulfonate (PSS) (PEDOT:PSS) for doping the organic channel. Atomic force microscopy (AFM) was used to obtain topographic maps of the gold gate. The electrochemical activity on gate electrodes was measured and related to the concentration of DNA extracted from samples and hybridized to the specific capture probe immobilized onto the gold surface of the gate. This assay reached a limit of detection of 1.05 ng/µL, corresponding to 0.56 pM of L. monocytogenes ATCC 7644, and allowed the specific and rapid detection of L. monocytogenes in the analyzed samples. KEYPOINTS: ⢠Textile organic electrochemical transistors functionalized with a specific DNA probe ⢠AFM topographic and surface potential maps of a functionalized gold gate surface ⢠Comparison between the Listeria monocytogenes Precis™ method and an OECT biosensor.
Assuntos
Técnicas Biossensoriais , Listeria monocytogenes , Listeria monocytogenes/genética , Técnicas Biossensoriais/métodos , Eletrodos , Alimentos , OuroRESUMO
Binary light-up aptamers are intriguing and emerging tools with potential in different fields. Herein, we demonstrate the versatility of a split Broccoli aptamer system able to turn on the fluorescence signal only in the presence of a complementary sequence. First, an RNA three-way junction harbouring the split system is assembled in an E. coli-based cell-free TX-TL system where the folding of the functional aptamer is demonstrated. Then, the same strategy is introduced into a 'bio-orthogonal' hybrid RNA/DNA rectangle origami characterized by atomic force microscopy: the activation of the split system through the origami self-assembly is demonstrated. Finally, our system is successfully used to detect the femtomoles of a Campylobacter spp. DNA target sequence. Potential applications of our system include the real-time monitoring of the self-assembly of nucleic-acid-based devices in vivo and of the intracellular delivery of therapeutic nanostructures, as well as the in vitro and in vivo detection of different DNA/RNA targets.
Assuntos
Aptâmeros de Nucleotídeos , Brassica , Nanoestruturas , RNA/genética , Brassica/genética , Escherichia coli/genética , Aptâmeros de Nucleotídeos/química , DNA/química , Nanoestruturas/química , Conformação de Ácido NucleicoRESUMO
This study reports the development of a sensitive magnetic bead-based enzyme-linked immunoassay (MELISA) for the pan-reactive detection of the Influenza A virus. The assay combines immunomagnetic beads and biotin-nanoparticle-based detection to quantify a highly conserved viral nucleoprotein in virus lysates. At the capture step, monoclonal antibody-coated magnetic microbeads were used to bind and concentrate the nucleoprotein in samples. The colorimetric detection signal was amplified using biotinylated silica nanoparticles (NP). These nanoparticles were functionalized on the surface with short DNA spacers bearing biotin groups by an automated supported synthesis method performed on nano-on-micro assemblies with a DNA/RNA synthesizer. A biotin-nanoparticle and immunomagnetic bead-based assay was developed. We succeeded in detecting Influenza A viruses directly in the lysis buffer supplemented with 10% saliva to simulate the clinical context. The biotin-nanoparticle amplification step enabled detection limits as low as 3 × 103 PFU mL-1 and 4 × 104 PFU mL-1 to be achieved for the H1N1 and H3N2 strains respectively. In contrast, a classical ELISA test based on the same antibody sandwich showed detection limit of 1.2 × 107 PFU mL-1 for H1N1. The new enhanced MELISA proved to be specific, as no cross-reactivity was found with a porcine respiratory virus (PRRSV). Graphical abstract.
Assuntos
Biotina/química , Separação Imunomagnética , Vírus da Influenza A/isolamento & purificação , Nanopartículas/química , Anticorpos Monoclonais , Sensibilidade e EspecificidadeRESUMO
The environmental contamination of soil by metal oxide nanomaterials is a growing global concern because of their potential toxicity. We investigated the effects of Mg doped ZnO (Mg-nZnO) nanoparticles on a model soil microorganism Bacillus subtilis. Mg-nZnO exhibited only a moderate toxic effect on B. subtilis vegetative cells but was able to prevent biofilm formation and destroy already formed biofilms. Similarly, Mg-nZnO (≤1â¯mg/mL) was moderately toxic towards Listeria monocytogenes, Staphylococcus aureus, Escherichia coli, Salmonella enterica, Saccharomyces cerevisiae and murine macrophages. Engineered Mg-nZnO produced H2O2 and O2â¢- radicals in solutions of various salt and organic molecule compositions. A quantitative proteomic analysis of B. subtilis membrane proteins showed that Mg-nZnO increased the expression of proteins involved in detoxification of ROS, translation and biofilm formation. Overall, our results suggest that Mg-nZnO released into the environment may hinder the spreading, colonization and biofilm formation by B. subtilis but also induce a mechanism of bacterial adaptation.
Assuntos
Bacillus subtilis/efeitos dos fármacos , Nanopartículas/toxicidade , Poluentes do Solo/toxicidade , Óxido de Zinco/toxicidade , Animais , Biofilmes , Escherichia coli/efeitos dos fármacos , Peróxido de Hidrogênio/metabolismo , Camundongos , Óxidos/metabolismo , Proteômica , Solo , Microbiologia do Solo , Staphylococcus aureusRESUMO
Foodborne pathogenic bacteria present a crucial food safety issue. Conventional diagnostic methods are time-consuming and can be only performed on previously produced food. The advancing field of point-of-need diagnostic devices integrating molecular methods, biosensors, microfluidics, and nanomaterials offers new avenues for swift, low-cost detection of pathogens with high sensitivity and specificity. These analyses and screening of food items can be performed during all phases of production. This review presents major developments achieved in recent years in point-of-need diagnostics in land-based sector and sheds light on current challenges in achieving wider acceptance of portable devices in the food industry. Particular emphasis is placed on methods for testing nucleic acids, protocols for portable nucleic acid extraction and amplification, as well as on the means for low-cost detection and read-out signal amplification.
Assuntos
Técnicas Biossensoriais/métodos , DNA Bacteriano/análise , Técnicas de Amplificação de Ácido Nucleico/métodos , Microbiologia de AlimentosRESUMO
BACKGROUND: Breast implants may be responsible for secondary deformities produced by parenchymal atrophy. However, few studies in the literature have reported changes in breast tissue after augmentation surgery. In this study, the breast thickness of patients undergoing breast augmentation was monitored by ultrasound, and correlations with surface, volume and projection measurements were examined. METHODS: We studied the parenchymal thickness at the lower pole of the breast with ultrasound in 36 women (72 breasts). In another group of 33 patients (66 breasts), we studied the thickness at the upper and lower poles along the meridian of each breast by ultrasound and measured the anthropometric metrics, volume and projection of the breast with a 3D camera. RESULTS: Midline measurements close to the areola showed reduced thickness at the lower pole, with 31.8% at the midpoint of the lower pole and 42% at the infra-areolar level (p < 0.0001). At the upper pole, there was a decrease of 14.6% (p < 0.001), but the thickness was increased by 6% and 38% at more cranial levels. No correlations with volume were found. Anatomical implants produced more atrophy at the lower pole, and round implants at the upper pole. More atrophy was found with subfascial than submuscular augmentation. Compared with the expected values, the final volume was very similar, but the projection was 29% less. Surface measurements changed significantly up to 4 months postoperatively and remained stable thereafter. CONCLUSIONS: Implants affect significatively the thickness of the glandular tissue. All changes occur very soon postoperatively but stabilize after 4 months. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .
Assuntos
Implantes de Mama , Glândulas Mamárias Humanas/efeitos dos fármacos , Glândulas Mamárias Humanas/diagnóstico por imagem , Géis de Silicone/farmacologia , Adulto , Feminino , Humanos , Imageamento Tridimensional , Pessoa de Meia-Idade , Desenho de Prótese , Ultrassonografia , Adulto JovemRESUMO
Infectious animal diseases caused by pathogenic microorganisms such as bacteria and viruses threaten the health and well-being of wildlife, livestock, and human populations, limit productivity and increase significantly economic losses to each sector. The pathogen detection is an important step for the diagnostics, successful treatment of animal infection diseases and control management in farms and field conditions. Current techniques employed to diagnose pathogens in livestock and poultry include classical plate-based methods and conventional biochemical methods as enzyme-linked immunosorbent assays (ELISA). These methods are time-consuming and frequently incapable to distinguish between low and highly pathogenic strains. Molecular techniques such as polymerase chain reaction (PCR) and real time PCR (RT-PCR) have also been proposed to be used to diagnose and identify relevant infectious disease in animals. However these DNA-based methodologies need isolated genetic materials and sophisticated instruments, being not suitable for in field analysis. Consequently, there is strong interest for developing new swift point-of-care biosensing systems for early detection of animal diseases with high sensitivity and specificity. In this review, we provide an overview of the innovative biosensing systems that can be applied for livestock pathogen detection. Different sensing strategies based on DNA receptors, glycan, aptamers and antibodies are presented. Besides devices still at development level some are validated according to standards of the World Organization for Animal Health and are commercially available. Especially, paper-based platforms proposed as an affordable, rapid and easy to perform sensing systems for implementation in field condition are included in this review.
Assuntos
Técnicas Biossensoriais/veterinária , Influenza Aviária/diagnóstico , Gado/microbiologia , Gado/virologia , Doenças das Aves Domésticas/diagnóstico , Animais , Técnicas Biossensoriais/métodos , Bluetongue/diagnóstico , Complexo Respiratório Bovino/diagnóstico , Infecções por Campylobacter/diagnóstico , Infecções por Campylobacter/veterinária , Bovinos , Galinhas/microbiologia , Galinhas/virologia , Infecções por Clostridium/diagnóstico , Infecções por Clostridium/veterinária , Coccidiose/diagnóstico , Coccidiose/virologia , Infecções por Escherichia coli/diagnóstico , Infecções por Escherichia coli/veterinária , Feminino , Febre Aftosa/diagnóstico , Mastite Bovina/diagnóstico , Infecções por Mycoplasma/diagnóstico , Infecções por Mycoplasma/veterinária , Infecções por Orthomyxoviridae/diagnóstico , Infecções por Orthomyxoviridae/veterinária , Salmonelose Animal/diagnósticoRESUMO
Speck is a meat product obtained from the deboned leg of pork that is salted, smoked and seasoned for four to six months. During speck seasoning, Eurotium rubrum and Penicillium solitum grow on the surface and collaborate with other moulds and tissue enzymes to produce the typical aroma. Both of these strains usually predominate over other moulds. However, moulds producing ochratoxins, such as Aspergillus ochraceus and Penicillium nordicum, can also co-grow on speck and produce ochratoxin A (OTA). Consequently, speck could represent a potential health risk for consumers. Because A. ochraceus and P. nordicum could represent a problem for artisanal speck production, the aim of this study was to inhibit these mould strains using Debaryomyces hansenii and Saccharomycopsis fibuligera. Six D. hansenii and six S. fibuligera strains were tested in vitro to inhibit A. ochraceus and P. nordicum. The D. hansenii DIAL 1 and S. fibuligera DIAL 3 strains demonstrated the highest inhibitory activity and were selected for in vivo tests. The strains were co-inoculated on fresh meat cuts for speck production with both of the OTA-producing moulds prior to drying and seasoning. At the end of seasoning (six months), OTA was not detected in the speck treated with both yeast strains. Because the yeasts did not adversely affect the speck odour or flavour, the strains are proposed as starters for the inhibition of ochratoxigenic moulds.
Assuntos
Antibiose , Aspergillus ochraceus/crescimento & desenvolvimento , Debaryomyces/crescimento & desenvolvimento , Produtos da Carne/microbiologia , Penicillium/crescimento & desenvolvimento , Carne Vermelha/microbiologia , Saccharomycopsis/crescimento & desenvolvimento , Animais , Aspergillus ochraceus/química , Aspergillus ochraceus/metabolismo , Agentes de Controle Biológico/metabolismo , Culinária , Debaryomyces/metabolismo , Microbiologia de Alimentos , Conservação de Alimentos/métodos , Produtos da Carne/análise , Ocratoxinas/análise , Ocratoxinas/biossíntese , Penicillium/química , Saccharomycopsis/metabolismo , SuínosRESUMO
In Friuli, a Northeastern region of Italy, a blood sausage called Sanganel is produced by farmers, butchers, shops, and factories. This sausage is made with pork meat, boiled blood, lard, spices, and salt. It is stored at 4 ± 2 °C and usually eaten fresh or boiled within 14 days of its manufacture. Little is known about its microbial populations and safety for consumption. The aim of this study is to characterise the microbial populations and the physico-chemical parameters of Sanganel to establish its quality and the safety of consuming it. The microbial population of Sanganel is typical of meat products, and psychrotrophic enterobacteria and lactic acid bacteria (LAB) grow while it is stored. Enterobacteria produce total basic volatile nitrogen (TVB-N) and biogenic amines that, despite the presence of LAB, increase the pH of the sausage to approximately 6.9. Considering the concentrations of Enterobacteriaceae and TVB-N in the sausage, a shelf-life of 14 days is suggested. However, at 30 days the sausage is safe to eat and presents normal odours and flavours. In addition, boiling the sausage for 30 min before consumption eliminates the asporogenous microbial population.
Assuntos
Bactérias/crescimento & desenvolvimento , Microbiologia de Alimentos , Armazenamento de Alimentos , Produtos da Carne/microbiologia , Consórcios Microbianos , Animais , Bactérias/isolamento & purificação , Aminas Biogênicas/análise , Contagem de Colônia Microbiana , Enterobacteriaceae/isolamento & purificação , Conservação de Alimentos , Inocuidade dos Alimentos , Concentração de Íons de Hidrogênio , Itália , Lactobacillaceae/isolamento & purificação , Produtos da Carne/análise , Refrigeração , SuínosRESUMO
Cooked bacon is a typical Italian meat product. After production, cooked bacon is stored at 4 ± 2 °C. During storage, the microorganisms that survived pasteurisation can grow and produce spoilage. For the first time, we studied the cause of the deterioration in spoiled cooked bacon compared to unspoiled samples. Moreover, the use of bio-protective cultures to improve the quality of the product and eliminate the risk of spoilage was tested. The results show that Leuconostoc mesenteroides is responsible for spoilage and produces a greening colour of the meat, slime and various compounds that result from the fermentation of sugars and the degradation of nitrogen compounds. Finally, Lactococcus lactis spp. lactis and Lactobacillus sakei were able to reduce the risk of Leuconostoc mesenteroides spoilage.
Assuntos
Embalagem de Alimentos , Conservação de Alimentos , Lactococcus lactis/fisiologia , Latilactobacillus sakei/fisiologia , Leuconostoc mesenteroides/crescimento & desenvolvimento , Produtos da Carne/microbiologia , Animais , Contagem de Colônia Microbiana , Fermentação , Contaminação de Alimentos , Microbiologia de Alimentos , SuínosRESUMO
The aim of this work was to determine the microorganisms present and to investigate their metabolites that cause spoilage of many goose sausages produced in Friuli, a northeast region of Italy. The defect was observed by sensorial analysis using the "needle probing" technique; the spoiled sausages were unsafe and not marketable. Despite the addition of starter, the microorganisms, particularly enterococci and Enterobacteriaceae, grew during ripening and produced a large amount of biogenic amines; therefore, these sausages represented a risk to consumers. The production of those compounds was confirmed in vitro. Furthermore, a second cause of spoilage was attributed to moulds that grew during ripening; the fungi grew between the meat and casing, producing a large amount of total volatile nitrogen, and consequently an ammonia smell was present either in the ripening area or in the sausages. This is the first description of this type of defect in goose sausages.
Assuntos
Aminas Biogênicas/metabolismo , Enterobacteriaceae/metabolismo , Enterococcus/metabolismo , Microbiologia de Alimentos , Fungos/metabolismo , Produtos da Carne/microbiologia , Animais , Aminas Biogênicas/análise , Cadaverina/metabolismo , Contagem de Colônia Microbiana , Enterobacteriaceae/crescimento & desenvolvimento , Enterococcus/crescimento & desenvolvimento , Fermentação , Inocuidade dos Alimentos , Fungos/crescimento & desenvolvimento , Gansos , Histamina/metabolismo , Itália , Nitrogênio/metabolismoRESUMO
Two recent genome-wide association studies in Asians have reported the association between the PSCA (prostate stem cell antigen) rs2294008C>T gene polymorphism and two Helicobacter pylori infection-related diseases such as gastric cancer (GC) and duodenal ulcer (DU). Since rs2294008 allele frequencies differ notably among ethnicities, we aimed to assess the role of rs2294008 on the susceptibility to GC and DU in a Caucasian population in Spain. Moreover, the relevance of rs2294008 on GC prognosis was evaluated. Genomic DNA from 603 Spanish patients with primary GC, 139 with DU and 675 healthy controls was typed for the PSCA rs2294008C>T polymorphism by PCR-TaqMan assays. H. pylori infection [odds ratio (OR): 8.27; 95% confidence interval (CI): 3.45-15.33] and nonsteroidal anti-inflammatory drugs (OR: 6.54; 95% CI: 3.19-12.43) were identified as independent risk factors for DU whereas the rs2294008T allele was associated with reduced risk of developing the disease (OR: 0.52; 95% CI: 0.33-0.82). Infection with CagA strains (OR: 2.10; 95% CI: 1.63-2.34), smoking (OR: 1.93; 95% CI: 1.54-2.61), family history of GC (OR: 2.83; 95% CI: 2.01-3.83), and the rs2294008T allele (OR: 1.46; 95% CI: 1.07-1.99) were associated with increased risk of GC. Interestingly, the association with the rs2294008T allele was restricted to noncardia GC (OR: 1.43; 95% CI: 1.12-1.82), particularly of the diffuse histotype (OR: 1.59; 95% CI: 1.16-1.92). Finally, Cox regression analysis identified the rs2294008T variant as a prognosis factor associated with worse overall survival in patients with diffuse-type GC (hazard ratio: 1.85; 95% CI: 1.12-3.06). From these results we conclude that the PSCA rs2294008 polymorphism is involved in the susceptibility to GC and DU, as well as in the prognosis of the diffuse-type of GC in Caucasians.
Assuntos
Antígenos de Neoplasias/genética , Úlcera Duodenal/genética , Predisposição Genética para Doença/genética , Proteínas de Neoplasias/genética , Polimorfismo Genético/genética , Neoplasias Gástricas/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Alelos , Estudos de Casos e Controles , Úlcera Duodenal/microbiologia , Úlcera Duodenal/patologia , Feminino , Proteínas Ligadas por GPI/genética , Estudo de Associação Genômica Ampla/métodos , Infecções por Helicobacter/genética , Infecções por Helicobacter/patologia , Helicobacter pylori , Humanos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Prognóstico , Risco , Fatores de Risco , Espanha , Neoplasias Gástricas/microbiologia , Neoplasias Gástricas/patologia , População Branca/genética , Adulto JovemRESUMO
The correct identification and characterisation of bacteria is essential for several reasons: the classification of lactic acid bacteria (LAB) has changed significantly over the years, and it is important to distinguish and define them correctly, according to the current nomenclature, avoiding problems in the interpretation of literature, as well as mislabelling when probiotic are used in food products. In this study, species-specific PCR and HRM (high-resolution melting) analysis were developed to identify strains belonging to the Lactobacillus casei group and to classify them into L. casei, Lactobacillus paracasei and Lactobacillus rhamnosus. HRM analysis confirmed to be a potent, simple, fast and economic tool for microbial identification. In particular, 201 strains, collected from International collections and attributed to the L. casei group, were examined using these techniques and the results were compared with consolidated molecular methods, already published. Seven of the tested strains don't belong to the L. casei group. Among the remaining 194 strains, 6 showed inconsistent results, leaving identification undetermined. All the applied techniques were congruent for the identification of the vast majority of the tested strains (188). Notably, for 46 of the strains, the identification differed from the previous attribution.
Assuntos
DNA Bacteriano/química , Lacticaseibacillus casei/isolamento & purificação , Reação em Cadeia da Polimerase Multiplex/métodos , Reação em Cadeia da Polimerase/métodos , DNA Bacteriano/genética , Lacticaseibacillus casei/classificação , Lacticaseibacillus casei/genética , Especificidade da Espécie , Temperatura de TransiçãoRESUMO
Adipocytolytic therapies have always raised the interest of aesthetic medicine physicians, mainly because of the great potential to achieve spectacular results in localized adiposities reduction. In the last few decades, these results have been severely compromised due to the improper or reckless injection of these products, to the extent of some of them being banned in many countries. Today, there is a new adipocytolytic solution that has been approved, is effective, and has theoretic and empiric consensus regarding its safety. The aim of this study for which 331 therapeutic sessions were retrospectively analyzed is to provide evidence of its safety and efficacy.
Assuntos
Tecido Adiposo/efeitos dos fármacos , Adiposidade/efeitos dos fármacos , Técnicas Cosméticas , Adulto , Técnicas Cosméticas/efeitos adversos , Feminino , Humanos , Injeções , Masculino , Pessoa de Meia-Idade , Soluções Farmacêuticas , Estudos RetrospectivosRESUMO
Brettanomyces bruxellensis is a current problem in winemaking all over the world, and the question if B. bruxellensis has a positive or negative impact on wine is one of the most controversial discussions in the world. The presence of live B. bruxellensis cells represents the risk of growth and an increase in cell numbers, which is related to the potential production of volatile phenols. In this work, the optimisation of a PMA-quantitative PCR (qPCR) method to enumerate only viable cells was carried out using the standard strain B. bruxellensis DSMZ 70726. The obtained detection limits were 0.83 log CFU/mL in red wine, 0.63 log CFU/mL in white wine and 0.23 log CFU/mL in beer. Moreover, the quantification was also performed by Reverse Transcription quantitative PCR (RT-qPCR), and the results showed a higher detection limit for all of the trials.
Assuntos
Cerveja/microbiologia , Brettanomyces/crescimento & desenvolvimento , Contaminação de Alimentos/análise , Reação em Cadeia da Polimerase/métodos , Vinho/microbiologia , Azidas/química , Brettanomyces/genética , Brettanomyces/isolamento & purificação , Viabilidade Microbiana , Reação em Cadeia da Polimerase/instrumentação , Propídio/análogos & derivados , Propídio/químicaRESUMO
The detection of Campylobacter, the most commonly reported cause of foodborne gastroenteritis in the European Union, is very important for human health. The most commonly recognised risk factor for infection is the handling and/or consumption of undercooked poultry meat. The methods typically applied to evaluate the presence/absence of Campylobacter in food samples are direct plating and/or enrichment culture based on the Horizontal Method for Detection and Enumeration of Campylobacter spp. (ISO 10272-1B: 2006) and PCR. Molecular methods also allow for the detection of cells that are viable but cannot be cultivated on agar media and that decrease the time required for species identification. The current study proposes the use of two molecular methods for species identification: dot blot and PCR. The dot blot method had a sensitivity of 25 ng for detection of DNA extracted from a pure culture using a digoxigenin-labelled probe for hybridisation; the target DNA was extracted from the enrichment broth at 24 h. PCR was performed using a pair of sensitive and specific primers for the detection of Campylobacter jejuni and Campylobacter coli after 24 h of enrichment in Preston broth. The initial samples were contaminated by 5 × 10 C. jejuni cells/g and 1.5 × 10(2)C. coli cells/g, thus the number of cells present in the enrichment broth at 0 h was 1 or 3 cell/g, respectively.