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BACKGROUND: To accurately measure ultrathin Descemet stripping automated endothelial keratoplasty (DSAEK) donor lamella thickness during the first postoperative year and to correlate this with pre-operative and other postoperative measurements. METHODS: Donor lamella thickness in 41 eyes undergoing DSAEK for Fuchs endothelial dystrophy (FED) was measured using the Tomey Casia OCT directly after graft preparation and at 1 week and 1, 3, 6 and 12 months postoperatively. Visual acuity and endothelial cell density were measured as the secondary parameters. RESULTS: Individual graft thickness profiles were shown to be fairly regular within the optically relevant area. There was a strong and highly significant correlation between the pre- and postoperative lamellar thicknesses at all time points (p < 0.0001). Compared with the measurements directly after preparation at the cornea bank, the lamella thickness decreased by 12% after 12 months. Between 1 and 12 months postoperatively, the lamella thickness (mean ± SD) changed from 112 ± 27 µm to 101 ± 21 µm. Best spectacle-corrected visual acuity (BSCVA) changed from 0.46 ± 0.30 logMAR pre-operatively through 0.36 ± 0.33 at 1 month to 0.13 ± 0.16 at 1 year postoperatively. The endothelial cell counts were comparable to those reported in previous studies. CONCLUSIONS: Thickness profiles of individual grafts were fairly regular within the optically relevant area. A strong relationship between pre- and postoperative graft thicknesses was detected, and ultrathin DSAEK grafts prepared using methods similar to that applied in this study are expected to show a deswelling of around 12% during the first postoperative year. No correlation was detected between graft thickness and BSCVA.
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Transplante de Córnea , Distrofia Endotelial de Fuchs , Humanos , Doadores de Tecidos , Córnea , Acuidade VisualRESUMO
OBJECTIVES: To evaluate radiographic bone level (RxBL) at dental implants and its associated factors in Spain. MATERIAL AND METHODS: This cross-sectional study was performed by a network of sentinel dentists from regions of Spain. RxBL was defined as the distance from the implant shoulder to the first clearly visible contact between the implant surface and the bone. Radiographic measurements were performed by two trained and experienced periodontists. Implant and patient data were also collected. Descriptive, bivariate, discriminative and multivariate analyses were done. RESULTS: A total of 49 sentinel dentists provided data 275 patients. Mean RxBL from 474 implants (5-13 years) was 1.87 mm (range: 0.00-13.17 mm). Statistically significant associations between RxBL and clinical output variables (bleeding on probing, oedema, plaque, probing depth, suppuration, keratinized tissue) were found. In the multiple regression analysis, statistically significant associations for RxBL were found for smoking habit, implant diameter, years of follow-up and type of prosthesis (p < 0.01). CONCLUSIONS: Peri-implant RxBL ranged from 0 to 13.17 mm. It was significantly associated with clinical output variables and with some potentially predictor variables, at patient- (smoking >10 cigarettes/day) and implant- (diameter, years of follow-up, Toronto bridge) levels.
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Perda do Osso Alveolar , Implantes Dentários , Placa Dentária , Peri-Implantite , Perda do Osso Alveolar/diagnóstico por imagem , Estudos Transversais , Humanos , EspanhaRESUMO
BACKGROUND: Infectious keratitis is the main cause of preventable blindness worldwide, with about 1.5-2.0 million new cases occurring per year. This inflammatory response may be due to infections caused by bacteria, fungi, viruses or parasites. Fungal keratitis is a poorly studied health problem. OBJECTIVES: This study aimed to identify a new fungal species by molecular methods and to explore the possible efficacy of the three most common antifungals used in human keratitis in Mexico by performing in vitro analysis. The capacity of this pathogen to cause corneal infection in a murine model was also evaluated. METHODS: The fungal strain was isolated from a patient with a corneal ulcer. To identify the fungus, taxonomic and phylogenetic analyses (nrDNA ITS and LSU data set) were performed. An antifungal susceptibility assay for amphotericin B, itraconazole and voriconazole was carried out. The fungal isolate was used to develop a keratitis model in BALB/c mice; entire eyes and ocular tissues were preserved and processed for histopathologic examination. RESULTS AND CONCLUSION: This fungal genus has hitherto not been reported with human keratitis in Mexico. We described a new species Purpurecillium roseum isolated from corneal infection. P roseum showed resistance to amphotericin B and itraconazole and was sensitive to voriconazole. In vivo study demonstrated that P roseum had capacity to developed corneal infection and to penetrate deeper corneal tissue. The global change in fungal infections has emphasised the need to develop better diagnostic mycology laboratories and to recognise the group of potential fungal pathogens.
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Antifúngicos/uso terapêutico , Hypocreales/classificação , Hypocreales/efeitos dos fármacos , Hypocreales/isolamento & purificação , Ceratite/microbiologia , Idoso , Anfotericina B/uso terapêutico , Animais , Córnea , DNA Fúngico , Farmacorresistência Fúngica/efeitos dos fármacos , Feminino , Humanos , Hypocreales/patogenicidade , Itraconazol/uso terapêutico , Ceratite/tratamento farmacológico , Ceratite/patologia , México , Camundongos , Camundongos Endogâmicos BALB C , Testes de Sensibilidade Microbiana , Técnicas de Tipagem Micológica , Micoses/tratamento farmacológico , Micoses/microbiologia , Filogenia , Voriconazol/uso terapêuticoRESUMO
In Mexico little is known about high-altitude glacial psychrotolerant or psychrophilic fungal species, with most glacial fungi isolated from polar environments or Alpine glaciers. It has been documented that some of these species may play an important role in bioremediation of contaminated environments with heavy metals. In the present study, 75 fungi were isolated from glaciers in Citlaltépetl (5675 masl) and Iztaccíhuatl (5286 masl) volcanoes. Combining morphological characteristics and molecular methods, based on ITS rDNA, 38 fungi were partially identified to genus level, 35 belonging to Ascomycota and three to Mucoromycota. The most abundant genera were Cladosporium, followed by Alternaria and Sordariomycetes order. All isolated fungi were psychrotolerant, pigmented and resistant to different concentrations of Cr(III) and Pb(II), while none tolerated Hg(II). Fungi most tolerant to Cr(III) and Pb(II) belong to the genera Stemphylium, Cladosporium and Penicillium and to a lesser extent Aureobasidium and Sordariomycetes. To our knowledge, this is the first report on cultivable mycobiota richness and their Cr and Pb tolerance. The results open new research possibilities about fungal diversity and heavy metals myco-remediation. Extremophilic fungal communities should be further investigated before global warming causes permanent changes and we miss the opportunity to describe these sites in Mexico.
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Camada de Gelo , Altitude , Biodegradação Ambiental , Fungos , México , MicobiomaRESUMO
Biofilms are structures that confer adaptive ability to and facilitate the virulence of fungal pathogens. Certain multi-functional proteins have been shown to be involved in fungal pathogenesis and these proteins may also be implicated in biofilm formation. The aim of this study was to identify a fungal agent isolated from the human cornea, to analyze the ability of this organism to form biofilms in vitro and to investigate protein expression in this condition. The fungus was identified by phylogenetic inference analysis. Biofilm formation and structure were evaluated by colorimetric methods and by optical and electron microscopy. We also resolved proteins obtained from biofilms and planktonic cultures by two-dimensional gel electrophoresis and identified those proteins by mass spectrometry. The fungus was identified as Fusarium falciforme. Colorimetric analysis and microscopy revealed that the highest level of biofilm formation was obtained at a concentration of 1â¯×â¯106 conidia/mL with 96â¯h of incubation at 28⯰C. The biofilm architecture consisted of an extracellular matrix that embedded fungal filaments. We found nineteen proteins that were over-expressed in biofilms, as compared with planktonic cultures, and six proteins with unique expression in biofilms. Among the more abundant proteins identified were: transketolase, a putative antigen 1, enolase, phosphoglycerate kinase and ATP-citrate synthase. Some of these proteins are involved in basal metabolism, function as multi-functional proteins or have been described as potential virulence factors. We focused on the expression in biofilm of the enzyme, enolase, which was determined by real-time PCR. Our findings provide a perspective on the proteins associated with the formation of biofilms in vitro by an F. falciforme keratitis isolate.
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Biofilmes/crescimento & desenvolvimento , Proteínas Fúngicas/análise , Fusarium/química , Fusarium/crescimento & desenvolvimento , Proteoma/análise , Córnea/microbiologia , Eletroforese em Gel Bidimensional , Infecções Oculares Fúngicas/microbiologia , Fusariose/microbiologia , Fusarium/isolamento & purificação , Humanos , Ceratite/microbiologia , Espectrometria de MassasRESUMO
BACKGROUND: In recent years, human keratitis caused by fungal plant pathogens has become more common. Biofilm is a structure that confers adaptations and virulence to fungi in keratitis. Neoscytalidium spp. are phytopathogenic and recently have been recognised as a human pathogen, using biofilm formation as a virulence factor. OBJECTIVES: The aim of this study was isolation, identification (at the species level) and characterisation of a new fungal keratitis agent. PATIENTS/METHODS: The fungus was isolated from a 67-year-old male patient with a corneal ulcer. Biofilm formation and structure were evaluated by colorimetric methods and microscopy. To identify the fungus, morphological characteristics were examined and a phylogenetic analysis was performed. RESULTS AND CONCLUSIONS: We report the identification of a fungus, a member of the genus Neoscytalidium which is associated with human keratitis. Phylogenetic analysis and morphological observations on conidiogenous cells, which occur only in arthric chains in aerial mycelium and the coelomycetous synasexual morph is absent, identified a new species, Neoscytalidium oculus sp. nov. The fungus formed biofilm at a concentration of 1 × 106 conidia/mL, during 96 hours of incubation at 37°C, and also manifested haemolysis and melanin production. This is the first report in Latin America of a new species of Neoscytalidium from a clinical isolate has been identified.
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Ascomicetos/classificação , Ascomicetos/isolamento & purificação , Biofilmes/crescimento & desenvolvimento , Úlcera da Córnea/microbiologia , Micoses/microbiologia , Idoso , Ascomicetos/genética , Ascomicetos/crescimento & desenvolvimento , Úlcera da Córnea/patologia , Humanos , Masculino , Técnicas Microbiológicas , Microscopia , Micoses/patologia , FilogeniaRESUMO
AIM: To evaluate the long-term hard and soft tissue peri-implant tissue stability of bone-level implants using a different implant placement protocol (submerged versus transmucosal). MATERIALS AND METHODS: This study was partly a subset analysis of a multicentre study where in 40 patients, a single bone-level implant with platform switching and a conical implant-abutment interface was placed either submerged or transmucosal in non-molar sites. Changes in the peri-implant tissues between implant placement and 5 years were assessed clinically and radiologically. Patient-related outcomes were also recorded. RESULTS: Thirty patients completed the 5-year follow-up. Implant survival rate was 100%. The mean radiographic changes in crestal bone levels between baseline and 5 years were 0.59 (0.92) mm and 0.78 (1.03) mm for the submerged and the transmucosal groups, respectively. No statistical significant differences were found between the groups for any of the investigated variables. Peri-implantitis, defined as changes in the level of crestal bone of ≥2 mm together with bleeding on probing, was only diagnosed in one patient. Patients in both groups were highly satisfied with the treatment received. CONCLUSIONS: Bone-level implants with submerged or transmucosal healing protocols demonstrated similar outcomes after 5 years. Both protocols yielded optimal clinical and radiographic results when bone-level implants were placed in non-molar positions for single tooth replacement.
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Dente Suporte , Implantação Dentária Endóssea , Implantes Dentários para Um Único Dente , Prótese Dentária Fixada por Implante , Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mucosa Bucal , Estudos Prospectivos , Método Simples-Cego , Ferida Cirúrgica , Fatores de Tempo , Resultado do Tratamento , Cicatrização , Adulto JovemRESUMO
PURPOSE: Traditionally, preoperative posturing consisting of bed rest and positioning is prescribed to patients with macula-on retinal detachment (RD) to prevent RD progression and detachment of the fovea. Execution of such advice can be cumbersome and expensive. This study aimed to investigate if preoperative posturing affects the progression of RD. DESIGN: Prospective cohort study. PARTICIPANTS: Ninety-eight patients with macula-on RD were included. Inclusion criteria were volume optical coherence tomography (OCT) scans could be obtained with sufficient quality; and the smallest distance from the fovea to the detachment border was 1.25 mm or more. METHODS: Patients were admitted to the ward for bed rest in anticipation of surgery and were positioned on the side where the RD was mainly located. At baseline and before and after each interruption for meals or toilet visits, a 37°×45° OCT volume scan was performed using a wide-angle Spectralis OCT (Heidelberg Engineering, Heidelberg, Germany). The distance between the nearest point of the RD border and fovea was measured using a custom-built measuring tool. MAIN OUTCOME MEASURES: The RD border displacement and the average RD border displacement velocity moving toward (negative) or away (positive) from the fovea were determined for intervals of posturing and interruptions. RESULTS: The median duration of intervals of posturing was 3.0 hours (interquartile range [IQR], 1.8-14.0 hours; n = 202) and of interruptions 0.37 hours (IQR, 0.26-0.50 hours; n = 197). The median RD border displacement was 2 µm (IQR, -65 to +251 µm) during posturing and -61 µm (IQR, -140 to 0 µm) during interruptions, a statistically significant difference (P < 0.001, Mann-Whitney U test). The median RD border displacement velocity was +1 µm/hour (IQR, -21 to +49 µm/hour) during posturing and -149 µm/hour (IQR, -406 to +1 µm/hour) during interruptions, a statistically significant difference (P < 0.001). CONCLUSIONS: By making use of usual interruptions of preoperative posturing we were able to show, in a prospective and ethically acceptable manner, that RD stabilizes during posturing and progresses during interruptions in patients with macula-on RD. Preoperative posturing is effective in reducing progression of RD.
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Repouso em Cama , Fóvea Central/patologia , Postura , Cuidados Pré-Operatórios/métodos , Descolamento Retiniano/diagnóstico , Estudos de Coortes , Progressão da Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Descolamento Retiniano/cirurgia , Recurvamento da Esclera , Tomografia de Coerência Óptica , Acuidade Visual/fisiologiaRESUMO
BACKGROUND: Knee osteoarthritis is a disease that affects a third of the population over 65 years of age, and it is increasingly becoming a motive for consultation and a source of pain and disability. The gold standard surgical treatment is a total knee arthroplasty; however, 15% to 30% of patients who have undergone surgery continue to experience pain and functional limitation. METHODS: A double-blind, randomized clinical study compared neurolysis using traditional radiofrequency (RF) to local anesthetic and corticosteroid block of the superolateral, superomedial, and inferomedial branches of the knee genicular nerves in patients who had total knee arthroplasty but still experience pain. RESULTS: Twenty-eight patients, 14 on each treatment arm, were followed for over a 1-year period. A reduction in pain and significant joint function improvement during the first 3 to 6 months was shown, with similar results using both techniques. No adverse effects were noted. An improvement in both disability and quality of life was observed, as well as a reduction in the need for analgesics in both treatment groups. CONCLUSIONS: Further clinical trials need to be undertaken, with a larger sample size, in order to demonstrate the efficacy of this technique and to detect the possible appearance of any long-term adverse effects.
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Denervação/métodos , Bloqueio Nervoso/métodos , Osteoartrite do Joelho/cirurgia , Manejo da Dor/métodos , Idoso , Anestésicos Locais , Artroplastia do Joelho , Método Duplo-Cego , Feminino , Humanos , Articulação do Joelho/inervação , Masculino , Pessoa de Meia-Idade , Medição da DorRESUMO
STUDY QUESTION: Is the drug used for final oocyte maturation a factor in determining the prevalence of empty follicle syndrome (EFS)? SUMMARY ANSWER: The drug used for final oocyte maturation is not a factor in determining the prevalence of EFS among women unaffected by infertility. WHAT IS KNOWN ALREADY: Despite satisfactory follicular stimulation and adequate follicular development, cases of EFS, i.e. failure to recover any cumulus oocyte complex, have been reported both with hCG and GnRH agonist triggering. No standard management protocol has been proposed so far. STUDY DESIGN, SIZE, DURATION: Retrospective analysis of oocyte donation cycles performed between August 2006 and April 2013 in a large private fertility centre. PARTICIPANTS/MATERIALS, SETTING, METHODS: The analysis included 12 483 oocyte donation cycles of which 74 were EFS cycles. All cycles were triggered with either hCG or GnRH agonists. MAIN RESULTS AND THE ROLE OF CHANCE: There were no differences in the gonadotropic stimulation, pituitary suppression and triggering drug between cycles where oocytes were recovered successfully and EFS cycles. The total prevalence of EFS was 0.59%. Given the rarity of the syndrome, caution is advised when interpreting the analysis. LIMITATIONS, REASONS FOR CAUTION: The main limitation of this study is its retrospective nature. Although this is the largest analysis of EFS in donors reported so far, its statistical power is limited because the syndrome has a low incidence. In some cycles of EFS from 2006 to 2007 there is a lack of hormone data. WIDER IMPLICATIONS OF THE FINDINGS: Our findings may be generalized to oocyte donors and IVF patients younger than 35 years old, with cycles undergoing final maturation triggering with either hCG or GnRH agonists. The generalization cannot be extended to patients with an ovarian factor as the cause of their reproductive pathology. The theoretical aetiology of a temporary hypothalamic-pituitary hyposensitivity can explain the cycles where a rescue protocol with hCG has been successful. STUDY FUNDING/COMPETING INTERESTS: This work was supported in part by funding from Fundaciò EUGIN. The authors have no conflicts to declare. TRIAL REGISTRATION NUMBER: NA.
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Gonadotropina Coriônica/agonistas , Hormônio Liberador de Gonadotropina/agonistas , Doação de Oócitos , Doenças Ovarianas/epidemiologia , Indução da Ovulação/métodos , Adulto , Feminino , Humanos , Estudos Retrospectivos , SíndromeRESUMO
OBJECTIVES: Three-dimensional (3D) sonographically based automated volume calculation (SonoAVC; GE Healthcare, Zipf, Austria) is being introduced in folliculometry during ovarian stimulation; however, clear training assessments in this technique are lacking. The learning curve-cumulative summation (LC-CUSUM) test provides a quantitative tool to determine when a trainee has learned a procedure. The aim of this prospective study was to assess 3D SonoAVC LC-CUSUM curves in folliculometry. METHODS: Analyses were performed on 98 oocyte donors by capturing the ovarian image in 3D and applying the 3D SonoAVC software during ovarian stimulation cycles. Each patient was scanned by an expert operator and by a trainee. Independent LC-CUSUM tests for 4 follicular diameters tracked the competency of 3 trainees in 3D SonoAVC. RESULTS: We found that the numbers of sonographic examinations required by the 3 trainees to identify the correct number of follicles of 10 mm or larger were 38, 32, and 28, respectively; for follicles of 14 mm or larger, they were 29, 28, and 28; for follicles of 18 mm or larger, they were 24, 19, and 27; and for follicles of 21 mm or larger, they were 29, 19, and 24. CONCLUSIONS: A variable number of procedures are needed to reach proficiency in 3D SonoAVC, even for trained 2-dimensional sonographers. Assessment of learning curves should be implemented when incorporating 3D SonoAVC in reproduction units.
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Competência Clínica , Imageamento Tridimensional/métodos , Infertilidade Feminina/terapia , Curva de Aprendizado , Folículo Ovariano/diagnóstico por imagem , Indução da Ovulação/métodos , Ultrassonografia/métodos , Adulto , Feminino , Humanos , Aumento da Imagem/métodos , Interpretação de Imagem Assistida por Computador/métodos , Infertilidade Feminina/diagnóstico por imagem , Variações Dependentes do Observador , Doação de Oócitos , Reconhecimento Automatizado de Padrão/métodos , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Western flower thrips, Frankliniella occidentalis (Pergande), is an economically important pest all over the world. New products against thrips are necessary, as there are few effective compounds exhibiting cross-resistance among them. Lethal effects, cross-resistance, and baseline susceptibility to spirotetramat were evaluated in this study. A new bioassay method for testing thrips against spirotetramat was developed. Spirotetramat showed a significant mortality effect on larvae after 7 d of exposure, but a low effect was observed on adults. Baseline results for larval instars showed LC50 values ranging from 11.59 to 49.81 mg AI/liter, with a low natural variability (3.2-fold). Cross-resistance studies showed overlapping confidence limits of the LC50 values for laboratory-selected (against acrinathrin, methiocarb, formetanate, and spinosad) resistant and susceptible strains, and low resistance factors, from 0.5 to 1.9, suggesting no cross-resistance to conventional insecticides previously used. A slight ovicidal effect (21-40% reduction) was also detected. Despite presenting low effects on adults, spirotetramat showed high but slow efficacy on F. occidentalis larvae. Field populations in southeast Spain showed a consistent susceptibility to spirotetramat. Given the scarcity of effective products and the lack of cross-resistance to other insecticides, spirotetramat can be considered as a good chemical tool to control F. occidentalis.
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Compostos Aza/farmacologia , Inseticidas/farmacologia , Larva , Compostos de Espiro/farmacologia , Tisanópteros/efeitos dos fármacos , Animais , Feminino , Resistência a Inseticidas , Larva/efeitos dos fármacos , Larva/genética , Larva/crescimento & desenvolvimento , Óvulo/efeitos dos fármacos , Espanha , Tisanópteros/genética , Tisanópteros/crescimento & desenvolvimentoRESUMO
Phaseolotoxin is an antimetabolite toxin produced by diverse pathovars of Pseudomonas syringae which affects various plants, causing diseases of economic importance. Phaseolotoxin contributes to the systemic dissemination of the pathogen in the plant, therefore it is recognized as a major virulence factor. Genetic traits such as the Pht cluster, appear defining to the toxigenic strains phaseolotoxin producers. Extensive research has contributed to our knowledge concerning the regulation of phaseolotoxin revealing a complex regulatory network that involves processes at the transcriptional and posttranscriptional levels, in which specific and global regulators participate. Even more, significant advances in understanding how specific signals, including host metabolites, nutrient sources, and physical parameters such as the temperature, can affect phaseolotoxin production have been made. A general overview of the phaseolotoxin regulation, focusing on the chemical and physical cues, and regulatory pathways involved in the expression of this major virulence factor will be given in the present work.
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DIN 16MnCr5 is commonly used in mechanical engineering contact applications such as gears, joint parts, shafts, gear wheels, camshafts, bolts, pins, and cardan joints, among others. This study examined the microstructural and mechanical properties and tribological behavior of different surface treatments applied to DIN 16MnCr5 steel. The samples were hardened at 870 °C for 15 min and then quenched in water. The surface conditions evaluated were as follows: quenched and tempered DIN 16MnCr5 steel samples without surface treatments (control group), quenched and tempered DIN 16MnCr5 steel samples with gas-nitriding at 560 °C for 6 h, quenched and tempered DIN 16MnCr5 steel samples with pack boriding at 950 °C for 4 h, and quenched and tempered DIN 16MnCr5 steel samples with duplex gas-nitriding and pack boriding. Microstructure characterization was carried out using metallographic techniques, optical microscopy, scanning electron microscopy with energy-dispersive spectroscopy, and X-ray diffraction. The mechanical properties were assessed through microhardness and elastic modulus tests using nanoindentation. The tribological behavior was evaluated using pin-on-disc tests following the ASTM G99-17 standard procedure under dry sliding conditions. The results indicated that the surface treated with duplex gas-nitriding and pack boriding exhibited the highest wear resistance and a reduced coefficient of friction due to improved mechanical properties, leading to increased hardness and elastic modulus.
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Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae), also known as the fall armyworm, is an economically important and widespread polyphagous pest. Microorganisms associated to this insect during life cycle play important ecological roles. We report 3 metagenome-assembled bacterial genomes reconstructed from a metagenome dataset obtained from S. frugiperda larvae F3 3rd-instar reared using artificial diet under laboratory conditions. Genome data for Enterococcus casseliflavus indicated a genome length of 3,659,8333 bp and GC content of 42.54%. Genome data for E. mundtii indicated a genome length of 2,921,701 bp and GC content of 38.37%. Finally, genome data for Lactiplantibacillus plantarum indicated a genome length of 3,298,601 bp, GC content of 44.31%. Genome analysis allowed us to identify genus-specific protein families (PLFams), transporters and antibiotic resistance-related genes among others. DNA sequences were deposited in National Center for Biotechnology Information (https://www.ncbi.nlm.nih.gov/) as Bioproject accession PRJNA899064.
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In this study, seed-surface-associated bacteria from fresh fruits of Capsicum spp. were analyzed to explore potential isolates for biocontrol of phytopathogenic fungal strains. A total of 76 bacterial isolates were obtained from three different species of chili pepper (C. annuum L., C. pubescens R. & P., and C. chinense Jacq.), and two isolates were selected via mycelial growth inhibition assays based on their production of volatile organic compounds (VOCs) against six fungal strains. Genomic analysis identified these isolates as Bacillus altitudinis CH05, with a chromosome size of 3,687,823 bp and with 41.25% G+C, and Bacillus tropicus CH13, with a chromosome size of 5,283,706 bp and with 35.24% G+C. Both bacterial strains showed high mycelial growth inhibition capacities against Sclerotium rolfsii, Sclerotinia sp., Rhizoctonia solani, and Alternaria alternata but lower inhibition capacities against Colletotrichum gloesporoides and Fusarium oxysporum. VOC identification was carried out after 24 h of fermentation with 64 VOCs for B. altitudinis CH05 and 53 VOCs for B. tropicus CH13. 2,5-Dimethyl pyrazine and acetoin had the highest relative abundance values in both bacterial strains. Our findings revealed that seed-surface-associated bacteria on Capsicum spp. have the metabolic ability to produce VOCs for biocontrol of fungal strains and have the potential to be used in sustainable agriculture.
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The competitive colonization of bacteria on similar ecological niches has a significant impact during their establishment. The synthesis speeds of different chemical classes of molecules during early competitive colonization can reduce the number of competitors through metabolic effects. In this work, we demonstrate for the first time that Kosakonia cowanii Cp1 previously isolated from the seeds of Capsicum pubescens R. P. produced volatile organic compounds (VOCs) during competitive colonization against Pectobacterium aroidearum SM2, affecting soft rot symptoms in serrano chili (Capsicum annuum L.). The pathogen P. aroidearum SM2 was isolated from the fruits of C. annuum var. Serrano with soft rot symptoms. The genome of the SM2 strain carries a 5,037,920 bp chromosome with 51.46% G + C content and 4925 predicted protein-coding genes. It presents 12 genes encoding plant-cell-wall-degrading enzymes (PCDEWs), 139 genes involved in five types of secretion systems, and 16 genes related to invasion motility. Pathogenic essays showed soft rot symptoms in the fruits of C. annuum L., Solanum lycopersicum, and Physalis philadelphica and the tubers of Solanum tuberosum. During the growth phases of K. cowanii Cp1, a mix of VOCs was identified by means of HS-SPME-GC-MS. Of these compounds, 2,5-dimethyl-pyrazine showed bactericidal effects and synergy with acetoin during the competitive colonization of K. cowanii Cp1 to completely reduce soft rot symptoms. This work provides novel evidence grounding a better understanding of bacterial interactions during competitive colonization on plant tissue, where VOC synthesis is essential and has a high potential capacity to control pathogenic microorganisms in agricultural systems.
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Chili powder is an important condiment around the world. However, according to various reports, the presence of pathogenic microorganisms could present a public health risk factor during its consumption. Therefore, microbiological quality assessment is required to understand key microbial functional traits, such as antibiotic resistance genes (ARGs). In this study, metagenomic next-generation sequencing (mNGS) and bioinformatics analysis were used to characterize the comprehensive profiles of the bacterial community and antibiotic resistance genes (ARGs) in 15 chili powder samples from different regions of Mexico. The initial bacterial load showed aerobic mesophilic bacteria (AMB) ranging between 6 × 103 and 7 × 108 CFU/g, sporulated mesophilic bacteria (SMB) from 4.3 × 103 to 2 × 109 CFU/g, and enterobacteria (En) from <100 to 2.3 × 106 CFU/g. The most representative families in the samples were Bacillaceae and Enterobacteriaceae, in which 18 potential pathogen-associated species were detected. In total, the resistome profile in the chili powder contained 68 unique genes, which conferred antibiotic resistance distributed in 13 different classes. Among the main classes of antibiotic resistance genes with a high abundance in almost all the samples were those related to multidrug, tetracycline, beta-lactam, aminoglycoside, and phenicol resistance. Our findings reveal the utility of mNGS in elucidating microbiological quality in chili powder to reduce the public health risks and the spread of potential pathogens with antibiotic resistance mechanisms.
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Electrostatic fermentation avoids the cellular redox imbalance of traditional fermentation, but knowledge gaps exist. This study explores the impact of electrostatic fermentation on the growth, volatile profile, and genetic response of Saccharomyces pastorianus Saflager S-23. The applied voltage (15 and 30 V) in the electrostatic fermentation system increased the growth and substrate utilization of S. pastorianus while decreasing ethanol production. The aromas typically associated with traditional fermentation, such as alcoholic, grape, apple, and sweet notes, were diminished, while aromas like roses, fruits, flowers, and bananas were augmented in electrostatic fermentation. RNA-seq analysis revealed upregulation of genes involved in cell wall structure, oxidoreductase activity, and iron ion binding, while genes associated with protein synthesis, growth control, homeostasis, and membrane function were downregulated under the influence of applied voltage. The electrostatic fermentation system modulates genetic responses and metabolic pathways in yeast, rendering it a promising method for tailored beer production. Demonstrating feasibility under industrial-scale and realistic conditions is crucial for advancing towards commercialization.
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Introduction: A high prevalence of advanced breast cancer (BC) is a common scenario in Latin America. In Peru, the frequency of BC at Stages III/IV is ≈50% despite implementation of a programme for breast cancer screening (BCS) along the country. We carried out a study to assess the feasibility and develop an instrument to evaluate the knowledge, barriers and perception about BCS in a nationwide pilot study in Peru among candidates for BCS. Methods: We conducted a systematic review of 2,558 reports indexed in PubMed, Scopus, Web of Science, Medline-Ovid and EMBASE, regarding to our study theme. In total, 111 were selected and a 51-items survey was developed (eight items about sociodemographic characteristics). Patients were recruited in public hospitals or private clinics, in rural and urban areas of nine departments of Peru. Results: We surveyed 488 women from: Lima (150), Cajamarca (93), Ica (59), Arequipa (56), Loreto (48), Ancash (38), Junín (15), Puerto Maldonado (15) and Huancavelica (14); 27.9% of them were from rural areas. The mean of age was 53.3 years (standard deviation ± 9.1). Regarding education level, 29.8% had primary, 33.2% secondary and 37.0% higher education. In total, 28.7% of women did not know the term 'mammogram' and 47.1% reported never receiving a BCS (36.9% from urban and 73.5% from rural population). In women that underwent BCS, only 67% knew it is for healthy women. In total, 54.1% of patients had low levels of knowledge about risk factors for BC (i.e. 87.5% of women respond that injuries in the breast produce cancer). Cultural, economic and geographic barriers were significantly associated with having a mammogram where 56.9% of participants considered a cost ≤ 7 USD as appropriate. Mammogram was perceived as too painful for 54.9% of women. In addition, women with a self-perception of low-risk for BC and a fatalistic perception of cancer were less likely to have a BCS. Conclusion: We found that it is feasible to conduct a large-scale study in Peru. The results of this pilot study highlight an urgent need of extensive education and awareness about BCS in Peru.