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1.
Eur Spine J ; 33(3): 1001-1012, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38267733

RESUMO

PURPOSE: This study aimed to compare the complication rates and functional outcomes between patients with and without a history of spinal fusion undergoing THA. METHODS: A systematic search was conducted across PubMed, EMBASE, Scopus, and Cochrane databases. Studies that compared adults with and without a history of spinal fusion after primary THA were included. The methodological quality of the studies was evaluated using MINORS criteria. Meta-analyses were performed utilizing mean differences (MD), standardized mean differences (SMD), and odds ratios (OR), along with 95% confidence intervals (CI). RESULTS: Seventeen studies involving 1,789,356 patients (31,786 in the SF group and 1,757,570 in the Non-SF group) were analyzed. The spinal fusion group exhibited significantly higher rates of dislocation (OR 2.50, 95% CI 1.78-3.52), periprosthetic fracture (OR 1.96, 95% CI 1.39-2.77), overall complications (OR 1.73, 95% CI 1.10-2.71), and revision rates (OR 1.86, 95% CI 1.74-1.99). Furthermore, within the first three months, there was an increased risk of dislocation (OR 4.38, 95% CI 1.36-14.14) and revisions (OR 3.87, 95% CI 1.63-9.18). Longer spinal fusions were significantly associated with a higher risk of dislocations (OR 0.62, 95% CI 0.53-0.71). Additionally, prior spinal fusion was linked to higher levels of pain (SMD 0.11, 95% CI 0.02-0.19) and poorer functional outcomes (MD - 0.09, 95% CI - 0.18 to - 0.00). CONCLUSIONS: Patients with a history of spinal fusion undergoing THA exhibit increased complication rates, higher levels of pain, and greater functional limitations than those without prior fusion. These findings have significant clinical implications for optimizing perioperative care in high-risk patient populations.


Assuntos
Artroplastia de Quadril , Luxações Articulares , Fusão Vertebral , Adulto , Humanos , Artroplastia de Quadril/efeitos adversos , Fusão Vertebral/efeitos adversos , Luxações Articulares/etiologia , Fatores de Risco , Dor/etiologia
2.
Vet Sci ; 10(9)2023 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-37756086

RESUMO

African swine fever virus is considered an emerging virus that causes African swine fever, a disease characterised by high mortality and elevated transmission rates and that, as it is for most other viral diseases, cannot be treated with specific drugs. Effective and reliable detection of the virus is relevant to prevent uncontrolled contagion among boar populations and to reduce economic losses. Moreover, animal health laboratories are demanding standardisation, optimisation and quality assurance of the available diagnostic assays. In the present study, the ASFV MONODOSE dtec-qPCR kit was validated following the UNE-EN ISO/IEC 17025:2005 guidelines. Analytical validation terms include in silico and in vitro specificity, sensitivity, efficiency and reliability (repeatability/reproducibility). Diagnostic validation of the method was assessed through the analysis of a total of 181 porcine samples originating from six different matrix types doped with African swine fever virus DNA received from the European reference laboratory for African Swine Fever (INIA-CISA, Madrid, Spain): whole blood, blood serum, kidney, heart, liver and tonsil. Results agreed with those obtained from a reference detection method also based on real-time PCR, endorsed by WOAH, but the ASFV MONODOSE dtec-qPCR kit incorporates some technical innovations and improvements which may benefit end-users. This kit, available worldwide with full analytical and diagnostic validation, can recognise all known ASFV genotypes and brings additional benefits to the current qPCR technology.

3.
Diagnostics (Basel) ; 13(9)2023 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-37174951

RESUMO

Human mpox is caused by the Monkeypox virus, a microorganism closely related to the Variola virus, both belonging to the Orthopoxvirus genus. Mpox had been considered a rare disease until a global outbreak occurred in 2022. People infected with the virus present similar symptoms to patients suffering smallpox and other rash illnesses, hindering diagnosis. The WHO indicated that no commercial PCR or serology kits are currently widely available. In the present study, the MPXV MONODOSE dtec-qPCR kit was validated following guidelines of the UNE/EN ISO/IEC 17025:2005. The parameters evaluated for the acceptance of the assay were in silico and in vitro specificity, quantitative phase analysis, reliability, and sensitivity. The assay passed validation criteria and yielded an efficiency of 95.8%, high repeatability, reproducibility, and a Limit of Detection and Quantification of at least 10 copies. Results from the validation of the MPXV dtec-qPCR kit were satisfactory. The use of the MONODOSE format (dehydrated single PCR-tubes, ready to use) provided considerable advantages allowing the detection of the Monkeypox virus to be accurately achieved. This detection kit may be considered a reliable, fast, simple, and universally available option.

5.
Viruses ; 14(11)2022 11 21.
Artigo em Inglês | MEDLINE | ID: mdl-36423197

RESUMO

A considerable number of new SARS-CoV-2 lineages have emerged since the first COVID-19 cases were reported in Wuhan. As a few variants showed higher COVID-19 disease transmissibility and the ability to escape from immune responses, surveillance became relevant at that time. Single-nucleotide mutation PCR-based protocols were not always specific, and consequently, determination of a high number of informative sites was needed for accurate lineage identification. A detailed in silico analysis of SARS-CoV-2 sequences retrieved from GISAID database revealed the S gene 921 bp-fragment, positions 22784-23705 of SARS-CoV-2 reference genome, as the most informative fragment (30 variable sites) to determine relevant SARS-CoV-2 variants. Consequently, a method consisting of the PCR-amplification of this fragment, followed by Sanger's sequencing and a "single-click" informatic program based on a reference database, was developed and validated. PCR-fragments obtained from clinical SARS-CoV-2 samples were compared with homologous variant-sequences and the resulting phylogenetic tree allowed the identification of Alpha, Delta, Omicron, Beta, Gamma, and other variants. The data analysis procedure was automatized and simplified to the point that it did not require specific technical skills. The method is faster and cheaper than current whole-genome sequencing methods; it is available worldwide, and it may help to enhance efficient surveillance in the fight against the COVID-19 pandemic.


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , Filogenia , Genoma Viral , COVID-19/diagnóstico , COVID-19/epidemiologia , Pandemias , Reação em Cadeia da Polimerase
6.
Pathogens ; 11(11)2022 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-36365050

RESUMO

The species Aeromonas lusitana was first described in 2016 with five strains recovered from untreated water and vegetables from Portugal. Since then, no further records exist of this species. During a surveillance study on the presence of Aeromonas in fish farms in Mexico, a new strain (ESV-351) of the mentioned species isolated from a rainbow trout was recovered. It was identified because it clustered phylogenetically with the type strain of A. lusitana based on the analysis of the rpoD gene sequences. In the present study, phenotypic characteristics, antimicrobial resistance profiles, and the presence of putative virulence genes of this novel strain (ESV-351) were determined in parallel to the five isolates from the original species description. Phenotypic differential characteristics exhibited by A. lusitana ESV-351 depicted an evident similarity to the characteristics exhibited by the other evaluated strains. However, the novel strain was positive for the production of indole using conventional methods, while the rest of the strains, including the type strain, were negative for its production. Furthermore, intermediate resistance to ampicillin, amoxicillin-clavulanic acid and cephalothin was detected in both the novel and the type strain. Five different virulence-related genes were detected in the novel strain and in the previously described strains, with the type strain exhibiting the highest number of virulence-related genes. In addition to this, the genome of the novel strain (ESV-351) was sequenced and compared with the genomes from the type strain (A. lusitana CECT 7828T) and other Aeromonas spp. The genomic analysis defined Aeromonas tecta as the closest species to A. lusitana with a highly similar number of predicted proteins. The genomic size, the number of protein-encoding genes and the number of different tRNAs, among other characteristics, make it possible to propose that the ESV-351 strain could potentially have the capacity to adapt to different environments. Genome comparison of the ESV-351 strain with the type strain revealed that both possess a similar sequence of the citrate synthase gene. In addition to this finding, the chromosomal region containing the citrate synthase locus of the novel strain exhibits some similarity to the chromosomal region in the genome of the A. hydrophila type strain and other known human pathogens, such as Vibrio cholerae. This could suggest a possible virulence role for the citrate synthase gene in A. lusitana (ESV-351).

7.
Methods Mol Biol ; 2517: 43-51, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35674944

RESUMO

Candida auris is a multidrug-resistant pathogenic ascomycete yeast of increasing health concern. C. auris colonizes patient's skin and can persist for weeks on surfaces, so it can be transmitted within and between hospitals. The most common diagnostic platforms in microbiology use reference databases that have not yet incorporated C. auris, misidentifying it. This chapter describes how to detect C. auris by qPCR with the GPS™ CanAur MONODOSE dtec-qPCR Test (Alicante, Spain) in less than 45 min, using ready-to-use tubes with all the components dehydrated. This commercial kit was subjected to validation following the guidelines of the UNE-EN ISO/IEC 17025:2005 and French Standard NF T90-471:2010.


Assuntos
Candida auris , Candida , Antifúngicos , Candida/genética , Humanos , Reação em Cadeia da Polimerase em Tempo Real
8.
Arch Microbiol ; 204(7): 415, 2022 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-35737122

RESUMO

Some weeks after the first CoVID-19 outbreak, the World Health Organization published some real-time PCR (qPCR) protocols developed by different health reference centers. These qPCR designs are being used worldwide to detect SARS-CoV-2 in the population, to monitor the prevalence of the virus during the pandemic. Moreover, some of these protocols to detect SARS-CoV-2 have widely been applied to environmental samples for epidemiological surveillance purposes. In the present work, the specificity of these currently used RT-qPCR designs was validated in vitro using SARS-CoV-2 and highly related coronaviral genomic sequences and compared to performance of the commercially available GPS™ CoVID-19 dtec-RT-qPCR Test. Assays performed with SARS-CoV-2-related genomes showed positive amplification when using some of these qPCR methods, indicating they may give SARS-CoV-2 false positives. This finding may be particularly relevant for SARS-CoV-2 monitoring of environmental samples, where an unknown pool of phylogenetically close-related viruses may exist.


Assuntos
COVID-19 , SARS-CoV-2 , COVID-19/diagnóstico , Humanos , Pandemias , Reação em Cadeia da Polimerase em Tempo Real/métodos , SARS-CoV-2/genética , Sensibilidade e Especificidade
9.
Microorganisms ; 8(5)2020 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-32414206

RESUMO

Hepatitis E virus (HEV) is one of the causative agents of water-borne human viral hepatitis and considered in Europe an emerging zoonotic pathogen. Analysis of bottled water through a standard method validated for HEV can contribute towards the risk management of this hazard. Putting some recent reports by the European Food Safety Authority in place, this study aimed to assess the performance of the concentration and extraction procedures described in ISO 15216-1:2017 for norovirus and hepatitis A virus on HEV detection. Following the ISO recommendation, the bottled water samples were spiked using serially diluted HEV fecal suspensions together with mengovirus as process control and concentrated by filtration via positively charged nylon membranes. In order to extract viral RNA from the resulting concentrates, two different methods were compared in this study: The one recommended in the ISO norm, NucliSens® MiniMag® system (NS), and an alternative commercially available kit NucleoSpin®RNA virus kit (MN). Finally, three reverse transcription quantitative PCR (RT-qPCR) assays were used to quantify HEV titers. The evaluated procedures resulted in average HEV recoveries of 14.08 ± 4.90% and 3.58 ± 0.30% for the MN and NS methods, respectively. The limit of detection (LoD95%) was 1.25 × 104 IU/L for both extraction methods combined with the three RT-qPCR assays tested, with the exception of NS extraction coupled with RT-qPCR1 that showed a LoD95% of 4.26 × 103 IU/L. The method characteristics generated in this study support the limited suitability of the ISO 15216-1:2017 concentration procedure coupled with the evaluated RT-qPCR assays for detecting HEV in bottled water.

10.
J Microbiol Methods ; 168: 105786, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31770538

RESUMO

There is increasing interest in methodologies for the simultaneous concentration and detection of multiple targets in individual samples. The aim of this study was to investigate the potential presence of E. coli DNA in beef extract powder used as part of a procedure to concentrate water samples for the simultaneous detection of bacteria, viruses and protozoa. DNA from E. coli was detected in five out of six beef extract lots tested, demonstrating the limitations of its inclusion when being used in assays that will be used for the detection of E. coli in water samples. Further work is required to clarify if this phenomenon also occurs for other microorganisms of interest in water.


Assuntos
DNA Bacteriano/isolamento & purificação , Escherichia coli O157/genética , Reação em Cadeia da Polimerase em Tempo Real , Carne Vermelha , Microbiologia da Água , Animais , Bovinos , Contagem de Colônia Microbiana , DNA Bacteriano/genética , Manejo de Espécimes/métodos
11.
Mycoses ; 61(11): 877-884, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30059175

RESUMO

Candida auris is an emerging multidrug resistant pathogenic fungus that causes candidaemia with high mortality rates and exhibits the ability to persist within the hospital environment. Candida auris is phylogenetically closely related to Candida haemulonii, C. lusitaniae, C. pseudohaemulonii and C. duobushaemulonii and is frequently misidentified by commercial identification methods. In the present study, the GPS™ MONODOSE dtec-qPCR kit (dried single-dose PCR tubes) for the detection of C. auris was validated following the guidelines of the UNE/EN ISO/IEC 17025:2005, the French Standard NF T90-471:2010 and using fast-cycling protocols. Validation terms included in vitro specificity (inclusivity/exclusivity), quantitative phase analysis (10-106  standard DNA copies), reliability (repeatability/reproducibility) and sensitivity (detection/quantification limits). GPS™ dtec-qPCR kits passed validation with strict acceptance criteria (n ≥ 10 repetitions). In silico specificity was proven by the designer (GPS™ ). Experimental inclusiveness was achieved in two independent laboratories by testing strain JCM15448T and 117 clinical C. auris isolates from Asia, the Middle-East Africa, Latin America and Europe. Exclusiveness was evaluated with 25 strains of closely related Candida spp. Use of the MONODOSE format provided considerable advantages allowing the detection of C. auris to be accurately achieved in less than an hour. The GPS™ MONODOSE dtec-qPCR kit is ready to undergo clinical evaluation.


Assuntos
Candida/isolamento & purificação , Candidíase/microbiologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , África , Candida/classificação , Candida/genética , Candidíase/diagnóstico , Europa (Continente) , Humanos , Técnicas de Tipagem Micológica , Reprodutibilidade dos Testes
12.
PeerJ ; 6: e4974, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29910986

RESUMO

BACKGROUND: The "One Health" concept recognizes that human health and animal health are interdependent and bound to the health of the ecosystem in which they (co)exist. This interconnection favors the transmission of bacteria and other infectious agents as well as the flow of genetic elements containing antibiotic resistance genes. This problem is worsened when pathogenic bacteria have the ability to establish as biofilms. Therefore, it is important to understand the characteristics and behaviour of microorganisms in both planktonic and biofilms states from the most diverse environmental niches to mitigate the emergence and dissemination of resistance. METHODS: The purpose of this work was to assess the antibiotic susceptibility of four bacteria (Acinetobacter spp., Klebsiella pneumoniae, Pseudomonas fluorescens and Shewanella putrefaciens) isolated from wild animals and their ability to form biofilms. The effect of two antibiotics, imipenem (IPM) and ciprofloxacin (CIP), on biofilm removal was also assessed. Screening of resistance genetic determinants was performed by PCR. Biofilm tests were performed by a modified microtiter plate method. Bacterial surface hydrophobicity was determined by sessile drop contact angles. RESULTS: The susceptibility profile classified the bacteria as multidrug-resistant. Three genes coding for ß-lactamases were detected in K. pneumoniae (TEM, SHV, OXA-aer) and one in P. fluorescens (OXA-aer). K. pneumoniae was the microorganism that carried more ß-lactamase genes and it was the most proficient biofilm producer, while P. fluorescens demonstrated the highest adhesion ability. Antibiotics at their MIC, 5 × MIC and 10 × MIC were ineffective in total biofilm removal. The highest biomass reductions were found with IPM (54% at 10 × MIC) against K. pneumoniae biofilms and with CIP (40% at 10 × MIC) against P. fluorescens biofilms. DISCUSSION: The results highlight wildlife as important host reservoirs and vectors for the spread of multidrug-resistant bacteria and genetic determinants of resistance. The ability of these bacteria to form biofilms should increase their persistence.

13.
Vet Microbiol ; 213: 28-34, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29292000

RESUMO

Chlamydia abortus, like other members of the family Chlamydiaceae, have a unique intracellular developmental cycle that is characterized by its chronic nature. Infection of a flock can remain undetected for months, until abortion occurs the following reproductive season but, to date, neither the location nor the mechanisms that maintain this latent phase are fully understood. Studies have shown that IL-10 produced as a response to certain micro-organisms sustains the intracellular survival of pathogens and increases host susceptibility to chlamydial infections. In order to induce a sustained infection C. abortus, transgenic mice that constitutively express IL-10 were infected and the immunological mechanisms that maintain infection in these mice were compared with the mechanisms of a resistant wild-type mouse strain. Viable bacteria could be detected in different tissues of transgenic mice up to 28 days after infection, as analysed by bacterial isolation and immunohistochemistry. Chronic infection in these mice was associated with an impaired recruitment of macrophages, decreased iNOS activity at the site of infection and a more diffuse distribution of inflammatory cells in the liver. This murine model can be of great help for understanding the immunological and bacterial mechanisms that lead to chronic chlamydial infections.


Assuntos
Infecções por Chlamydia/microbiologia , Chlamydia/imunologia , Interleucina-10/genética , Animais , Embrião de Galinha , Infecções por Chlamydia/patologia , Doença Crônica , Modelos Animais de Doenças , Feminino , Interleucina-10/metabolismo , Macrófagos/microbiologia , Camundongos , Camundongos Transgênicos
14.
Curr Microbiol ; 72(6): 795-803, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26868258

RESUMO

During previous studies to evaluate the phylogenetic diversity of Aeromonas from untreated waters and vegetables intended for human consumption, a group of isolates formed a unique gyrB phylogenetic cluster, separated from those of all other species described so far. A subsequent extensive phenotypic characterization, DNA-DNA hybridization, 16S rRNA gene sequencing, multi-locus phylogenetic analysis of the concatenated sequence of seven housekeeping genes (gyrB, rpoD, recA, dnaJ, gyrA, dnaX, and atpD; 4705 bp), and ERIC-PCR, were performed in an attempt to ascertain the taxonomy position of these isolates. This polyphasic approach confirmed that they belonged to a novel species of the genus Aeromonas, for which the name Aeromonas lusitana sp. nov. is proposed, with strain A.11/6(T) (=DSMZ 24095(T), =CECT 7828(T)) as the type strain.


Assuntos
Aeromonas/isolamento & purificação , Água Doce/microbiologia , Verduras/microbiologia , Aeromonas/classificação , Aeromonas/genética , DNA Bacteriano/genética , DNA Ribossômico/genética , Contaminação de Alimentos/análise , Humanos , Filogenia , RNA Ribossômico 16S/genética
15.
J Health Econ Outcomes Res ; 4(1): 35-46, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-34430666

RESUMO

Background: Anti-thrombotic prophylaxis is routinely used in patients undergoing elective total hip or knee replacement (THR or TKR) to reduce the risk of venous thromboembolism (VTE). In Spain, pharmacological prophylaxis is performed with low-molecular-weight heparin, enoxaparin being the most commonly used. Rivaroxaban is an oral antithrombotic drug that has shown superior efficacy and similar safety profile compared to enoxaparin regimens in randomized clinical trials. The aim of the study was to estimate the budget impact of increasing the use of rivaroxaban with respect to enoxaparin in the prophylaxis of VTE in patients undergoing elective THR or TKR. Methods: A budget impact analysis was conducted in order to estimate the economic cost from an increase of rivaroxaban use versus enoxaparin by 10%, 20%, and 30% over the 3 years of the time horizon (2015, 2016, and 2017) for the THR and TKR populations. Data related to rate of thromboembolic events, major bleeding events and use of resources (local or general anesthesia and nurse care after surgery) were obtained from the Xarelto® for VTE Prophylaxis After Hip or Knee Arthroplasty (XAMOS) study, an international, non-interventional, observational, open-label study in unselected patients undergoing THR or TKR surgery in routine practice. The study included a total of 17 701 patients from 252 centers in 37 countries, including Spain, Italy, France and United Kingdom, among others. Two cohorts where considered (patients undergoing THR or TKR) with two arms (patients treated with rivaroxaban or enoxaparin). The Spanish patients enrolled in the XAMOS study were 262 with THR and 538 with TKR. Thromboembolic events, major bleeding rates and health care resources were considered from both the international and the Spanish population. Health care resources including pharmacologic prophylaxis, anesthesia and nurse care costs (Euros 2014) were estimated from the Spanish National Healthcare System (NHS) perspective. The annual cost associated with each cohort was estimated based on the mean cost per patient and the estimated distribution of use of rivaroxaban or enoxaparin in the base case scenario and alternative scenario (increase of rivaroxaban use) over the 3 years. A one-way sensitivity analysis was conducted to evaluate the effect that the uncertainty of the input parameters may have on the results of the impact budget. Results: The difference in cost per patient undergoing THR or TKR with rivaroxaban versus enoxaparin was -€140.69 including event rates and resource use from the Spanish XAMOS population, and -€110.54 when considering event rates and resource use from the multinational XAMOS population (including but not limited to European [Spain, France, Italy, United Kingdom, Portugal, etc.], American [Canada, Mexico, Colombia, Venezuela, etc.], Asian [China, etc.] and Australian countries). In the analysis per cohort (THR or TKR), the impact of increasing the use of rivaroxaban in the THR cohort, was -€1106, -€2875, and -€5607 for 2015, 2016, and 2017, considering the data from the Spanish XAMOS population, and -€869, -€2259, and -€4405 considering the data from the multinational population. Considering the TKR cohort, the impact was -€2271, -€5904, and -€11 513, and -€1784, €4639, and -€9046, respectively. Conclusions: The present analysis shows that, according to effectiveness data from the XAMOS study (Spanish and multinational cohorts), an increase in the usage of rivaroxaban in VTE prophylaxis would lead to significant direct cost reduction in elective THR and TKR patients.

16.
Syst Appl Microbiol ; 38(1): 30-5, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25497285

RESUMO

Two strains recovered from mussels (F128-2(T)) and sea water (W63(T)) were characterized as Arcobacter sp., but they could not be assigned to any known species using the molecular identification methods specific for this genus (16S rDNA-RFLP and m-PCR) and rpoB gene analysis. The 16S rRNA gene sequence similarity to the type strains of all Arcobacter species ranged from 92.2% to 96.7% with strain F128-2(T), and from 94.1% to 99.4% with strain W63(T), the most similar being A. bivalviorum (CECT 7835(T)) and A. defluvii (CECT 7697(T)), respectively. The phylogenetic analyses of 16S rRNA, and the concatenated sequences of gyrB, gyrA, rpoB, atpA and hsp60 genes confirmed that strains F128-2(T) and W63(T) belonged to two new lineages within the genus Arcobacter. Moreover, both strains showed differential phenotypic characteristics and MALDI-TOF mass spectra from all other Arcobacter species. Therefore, it has been demonstrated the existence of two new Arcobacter species and the proposed names are Arcobacter ebronensis (type strain F128-2(T)=CECT 8441(T)=LMG 27922(T)), and Arcobacter aquimarinus (type strain W63(T)=CECT 8442(T)=LMG 27923(T)).


Assuntos
Arcobacter/isolamento & purificação , Microbiologia da Água , DNA Bacteriano/genética , Dados de Sequência Molecular , Tipagem Molecular , Filogenia , RNA Ribossômico 16S/genética , Água do Mar/microbiologia , Análise de Sequência de DNA
17.
Connect Tissue Res ; 55(3): 197-204, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24432912

RESUMO

Acid-sensing ion channels (ASICs) are a family of H(+)-gated voltage-insensitive ion channels that respond to extracellular acidification by regulating transmembrane Ca(2+) flux. Moreover, ASICs can also be gated by mechanical forces and may function as mechanosensors. The cells of the intervertebral disc (IVD) have an unusual acidic and hyperosmotic microenvironment. Changes in the pH and osmolarity determine the viability of IVD cells and the composition of the extracellular matrix, and both are the basis of IVD degeneration. In this study, the expression of ASICs (ASIC1, ASIC2, ASIC3 and ASIC4) mRNAs and proteins in human healthy and degenerated IVD was evaluated by quantitative reverse transcription-quantitative polymerase chain reaction and Western blot. The distribution of ASIC proteins was determined by immunohistochemistry. The mRNAs for all ASICs were detected in normal human IVD, and significantly increased levels were found in degenerated IVD. Western blots demonstrated the presence of proteins with estimated molecular weights of approximately 68-72 kDa. In both the annulus fibrosus (AF) and nucleus pulposus (NP) of normal IVD, ASIC2 is the most frequently expressed ASIC followed by ASIC3, ASIC1 and ASIC4. In the AF of degenerated IVD, there was a significant increase in the number of ASIC1 and ASIC4 positive cells, whereas in the NP, we found significant increase of expression of ASIC1, ASIC2 and ASIC3. These results describe the occurrence and localization of different ASICs in human healthy IVD, and their increased expression in degenerated IVD, thus suggesting that ASICs may be involved in IVD degeneration.


Assuntos
Canais Iônicos Sensíveis a Ácido/metabolismo , Degeneração do Disco Intervertebral/metabolismo , Disco Intervertebral/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Adulto , Idoso , Cálcio/metabolismo , Matriz Extracelular/metabolismo , Feminino , Humanos , Imuno-Histoquímica/métodos , Disco Intervertebral/patologia , Masculino , Proteínas de Membrana/metabolismo , Pessoa de Meia-Idade
18.
Clin Orthop Relat Res ; 471(11): 3543-53, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23884802

RESUMO

BACKGROUND: Porous tantalum is an option of cementless fixation for TKA, but there is no randomized comparison with a cemented implant in a mid-term followup. QUESTIONS/PURPOSES: We asked whether a tibial component fixed by a porous tantalum system might achieve (1) better clinical outcome as reflected by the Knee Society Score (KSS) and WOMAC Osteoarthritis Index, (2) fewer complications and reoperations, and (3) improved radiographic results with respect to aseptic loosening compared with a conventional cemented implant. METHODS: We randomized 145 patients into two groups, either a porous tantalum cementless tibial component group (Group 1) or cemented conventional tibial component in posterior cruciate retaining TKA group (Group 2). Patients were evaluated preoperatively and 15 days, 6 months, and 5 years after surgery, using the KSS and the WOMAC index. Complications, reoperations, and radiographic failures were tallied. RESULTS: At 5-year followup the KSS mean was 90.4 (range, 68-100; 95% CI, ± 1.6) for Group 1, and 86.5 (range, 56-99; 95% CI, ± 2.4) for Group 2. The effect size, at 95% CI for the difference between means, was 3.88 ± 2.87. The WOMAC mean was 15.1 (range, 0-51; 95% CI, ± 2.6) for the Group 1, and 19.1 (range, 4-61; 95% CI, ± 2.9) for Group 2. The effect size for WOMAC was -4.0 ± 3.9. There were no differences in the frequency of complications or in aseptic loosening between the two groups. CONCLUSIONS: Our data suggest there are small differences between the uncemented porous tantalum tibial component and the conventional cemented tibial component. It currently is undetermined whether the differences outweigh the cost of the implant and the results of their long-term performance.


Assuntos
Artroplastia do Joelho/instrumentação , Articulação do Joelho/cirurgia , Prótese do Joelho , Tantálio , Idoso , Artroplastia do Joelho/efeitos adversos , Fenômenos Biomecânicos , Cimentos Ósseos/uso terapêutico , Distribuição de Qui-Quadrado , Feminino , Humanos , Articulação do Joelho/diagnóstico por imagem , Articulação do Joelho/fisiopatologia , Masculino , Pessoa de Meia-Idade , Porosidade , Complicações Pós-Operatórias/etiologia , Complicações Pós-Operatórias/cirurgia , Desenho de Prótese , Falha de Prótese , Radiografia , Recuperação de Função Fisiológica , Reoperação , Fatores de Risco , Espanha , Fatores de Tempo , Resultado do Tratamento
19.
Syst Appl Microbiol ; 36(3): 171-6, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23485124

RESUMO

Previous studies indicate that Aeromonas aquariorum and Aeromonas hydrophila subsp. dhakensis are the same taxon and suggest that they should be synonymized. Using a polyphasic approach, the phenotypic and phylogenetic relationship of A. aquariorum with the 3 defined A. hydrophila subspecies (i.e. dhakensis, hydrophila, ranae) was investigated. Phylogenetic trees derived from the 16S rRNA, rpoD or gyrB genes and a multilocus phylogenetic analysis (with the concatenated sequences of gyrB, rpoD, recA, dnaJ and gyrA) confirmed that both A. aquariorum and A. hydrophila subsp. dhakensis are a unique taxon, different from the other A. hydrophila subspecies, corroborating the phenotypic and DNA-DNA hybridization (DDH) results. A formal synonymization of A. aquariorum and A. hydrophila subsp. dhakensis and a reclassification of both as Aeromonas dhakensis sp. nov. comb nov. is therefore proposed.


Assuntos
Aeromonas/classificação , Aeromonas/genética , Aeromonas hydrophila/classificação , Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana , Genes Essenciais , Fenótipo , Filogenia , RNA Ribossômico 16S/genética
20.
Int J Syst Evol Microbiol ; 63(Pt 6): 2270-2276, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23159749

RESUMO

A Gram-negative, facultatively anaerobic bacillus, designated strain 266(T), was isolated from an irrigation water system in the south-west of Western Australia. Analysis of the 16S rRNA gene sequence confirmed that strain 266(T) belonged to the genus Aeromonas, with the nearest species being Aeromonas fluvialis (99.6% similarity to the type strain, with 6 nucleotide differences) followed by Aeromonas veronii and Aeromonas allosaccharophila (both 99.5%). Analysis of gyrB and rpoD sequences suggested that strain 266(T) formed a phylogenetic line independent of other species in the genus. This was confirmed using the concatenated sequences of six housekeeping genes (gyrB, rpoD, recA, dnaJ, gyrA and dnaX) that also indicated that A. veronii and A. allosaccharophila were the nearest relatives. DNA-DNA reassociation experiments and phenotypic analysis further supported the conclusion that strain 266(T) represents a novel species, for which the name Aeromonas australiensis sp. nov. is proposed, with type strain 266(T) (=CECT 8023(T) =LMG 26707(T)). [corrected].


Assuntos
Aeromonas/classificação , Filogenia , Microbiologia da Água , Aeromonas/genética , Aeromonas/isolamento & purificação , Irrigação Agrícola , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Ácidos Graxos/análise , Genes Bacterianos , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Austrália Ocidental
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