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1.
Eur J Clin Microbiol Infect Dis ; 43(4): 673-682, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38296911

RESUMO

PURPOSE: To investigate the occurrence of vancomycin-variable enterococci (VVE) in a hospital in central Italy. METHODS: vanA positive but vancomycin-susceptible Enterococcus faecium isolates (VVE-S) were characterized by antibiotic susceptibility tests, molecular typing (PFGE and MLST), and WGS approach. The reversion of VVE-S to a resistant phenotype was assessed by exposure to increasing vancomycin concentrations, and the revertant isolates were used in filter mating experiments. qPCR was used to analyze the plasmid copy number. RESULTS: Eleven putative VVE-S were selected. WGS revealed two categories of vanA cluster plasmid located: the first type showed the lack of vanR, the deletion of vanS, and an intact vanH/vanA/vanX cluster; the second type was devoid of both vanR and vanS and showed a deletion of 544-bp at the 5'-end of the vanH. Strains (n = 7) carrying the first type of vanA cluster were considered VVE-S and were able to regain a resistance phenotype (VVE-R) in the presence of vancomycin, due to a 44-bp deletion in the promoter region of vanH/vanA/vanX, causing its constitutive expression. VVE-R strains were not able to transfer resistance by conjugation, and the resistance phenotype was unstable: after 11 days of growth without selective pressure, the revertants were still resistant but showed a lower vancomycin MIC. A higher plasmid copy number in the revertant strains was probably related to the resistance phenotype. CONCLUSION: We highlight the importance of VVE transition to VRE under vancomycin therapy resulting in a potential failure treatment. We also report the first-time identification of VVE-S isolates pstS-null belonging to ST1478.


Assuntos
Enterococcus faecium , Infecções por Bactérias Gram-Positivas , Humanos , Vancomicina/farmacologia , Vancomicina/uso terapêutico , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Tipagem de Sequências Multilocus , Resistência a Vancomicina/genética , Testes de Sensibilidade Microbiana , Enterococcus , Proteínas de Bactérias/genética , Infecções por Bactérias Gram-Positivas/microbiologia
2.
Environ Sci Technol ; 58(16): 6924-6933, 2024 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-38608723

RESUMO

Paralytic shellfish toxins (PSTs) produced by marine dinoflagellates significantly impact shellfish industries worldwide. Early detection on-farm and with minimal training would allow additional time for management decisions to minimize economic losses. Here, we describe and test a standardized workflow based on the detection of sxtA4, an initial gene in the biosynthesis of PSTs. The workflow is simple and inexpensive and does not require a specialized laboratory. It consists of (1) water collection and filtration using a custom gravity sampler, (2) buffer selection for sample preservation and cell lysis for DNA, and (3) an assay based on a region of sxtA, DinoDtec lyophilized quantitative polymerase chain reaction (qPCR) assay. Water samples spiked with Alexandrium catenella showed a cell recovery of >90% when compared to light microscopy counts. The performance of the lysis method (90.3% efficient), Longmire's buffer, and the DinoDtec qPCR assay (tested across a range of Alexandrium species (90.7-106.9% efficiency; r2 > 0.99)) was found to be specific, sensitive, and efficient. We tested the application of this workflow weekly from May 2016 to 30th October 2017 to compare the relationship between sxtA4 copies L-1 in seawater and PSTs in mussel tissue (Mytilus galloprovincialis) on-farm and spatially (across multiple sites), effectively demonstrating an ∼2 week early warning of two A. catenella HABs (r = 0.95). Our tool provides an early, accurate, and efficient method for the identification of PST risk in shellfish aquaculture.


Assuntos
Aquicultura , Dinoflagellida , Proliferação Nociva de Algas , Toxinas Marinhas , Fluxo de Trabalho , Animais , Frutos do Mar , Fazendas , Intoxicação por Frutos do Mar
3.
Endocr J ; 2024 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-38811206

RESUMO

Post-traumatic pituitary stalk transection syndrome (PSTS) is an extremely rare cause of combined pituitary hormone deficiency (CPHD), affecting approximately 9 per 100,000 cases of traumatic brain injury. In contrast, pituitary stalk interruption syndrome (PSIS) is also a rare cause of CPHD. Importantly, these conditions are often confused due to their similar names and resembling findings on magnetic resonance imaging (MRI). PSIS has been thought to be a prenatal developmental event resulting from a couple of genetic aberrations. In typical PSIS, anterior pituitary hormone deficiencies are restricted to growth hormone (GH) and gonadotropin during the pediatric age, gradually and generally progressing to panhypopituitarism in most cases. In contrast, global deficiencies of the anterior pituitary hormones in PSTS are temporally associated with trauma. To the best of our knowledge, no case reports of PSTS combined with acute traumatic spinal cord injury have been reported. A 34-year-old female was transferred to our hospital after jumping from the fourth building floor. She was diagnosed as an acute traumatic spinal cord injury and underwent the operation of elective posterior spinal fusion. On postoperative day 7, the blood tests revealed considerable hyperkalemia, hyponatremia and eosinophilia. Notably, menstruation stopped after falling from a height. Pituitary function tests revealed GH deficiency, hypogonadism, hypothyroidism and hypoadrenocorticism. MRI revealed loss of the pituitary stalk, whilst the hyperintense signal from distal axon of hypothalamus was still identified. Based on these findings, she was diagnosed as PSTS. Our case highlights endocrinological landscape of transection of the pituitary stalk by acute trauma.

4.
BMC Musculoskelet Disord ; 24(1): 720, 2023 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-37689657

RESUMO

OBJECTIVES: This study aimed to analyze the change trends of prevertebral soft tissue swelling (PSTS) for anterior cervical corpectomy fusion(ACCF) and to evaluate the risk factors of PSTS for postoperative dysphagia. METHODS: There were 309 patients with degenerative cervical diseases who were treated with ACCF from November 2015 and September 2019 in our hospital. According to the symptom of swallowing function after ACCF, those were divided into the dysphagia group and the normal-swallowing function group. Cervical computed tomography(CT) was analyzed, and radiological evaluation of the prevertebral soft tissue was measured between the antero-inferior corner of each vertebral body and the air shadow of the airway through CT mid-sagittal slice images before operation and after operation(one week, one month, eight months and twelve months). RESULTS: The incidence of dysphagia after ACCF was 41.1%. 120 of 127(94.5%) patients had dysphagia disappeared at the 8 months after ACCF, and all disappeared at the 12 months. In both groups, PSTS would be biggest at 1 week postoperatively comparing to the preoperative, and then get smaller from 1 week to 12 months postoperatively (p < 0.05). After 12 months of operation, the PSTS of all cervical spinal levels would get equal to the preoperative size in the normal-swallowing function group, while the PSTS in dysphagia group would get equal only in C5-7 levels. The PSTS of preoperative C6 level and postoperative C2 level were more closely related to the present of postoperative dysphagia (OR: 9.403, 95%CI: 2.344-37.719, OR: 3.187, 95%CI: 1.78-5.705). It was more important to predict postoperative dysphagia using the value of PSTS at preoperative C6 level and postoperative C2 level, with the cutoff threshold for the PSTS of preoperative C6 level ≦1.51 cm and postoperative C2 level ≦1.3915 cm, which could get sensitivity & specificity 66.929% and 61.54%, 77.17% and 64.29%, respectively. CONCLUSION: Our study showed that the increasing of the PSTS after ACCF should be considered as a risk factor of dysphagia after surgery. With the recovery of PSTS over time, the incidence of postoperative dysphagia decreases. The PSTS of preoperative C6 level and and postoperative C2 level should play an important part in predicting the risk of postoperative dysphagia.


Assuntos
Transtornos de Deglutição , Humanos , Transtornos de Deglutição/epidemiologia , Transtornos de Deglutição/etiologia , Fatores de Risco , Hospitais , Pescoço , Período Pós-Operatório
5.
Educ Inf Technol (Dordr) ; 28(5): 5739-5761, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36373042

RESUMO

During the COVID-19 pandemic, we founded an Online Laboratory School (OLS) under the roof of a university in Turkey to support students from public schools that were not technologically prepared for an online education and to provide an opportunity for our pre-service teachers (PSTs) to continue their internship by teaching online. The purpose of this research, consisting of two studies, was to examine experiences of 43 PSTs (first-, third- and fourth-years) during the OLS period of 8 weeks and how the OLS affected their mathematics teaching anxiety during Fall 2020. In the first study, we administered a survey to inquire into PSTs' views on their experiences at the OLS, and in the second study we examined their mathematics teaching anxiety before and after the OLS experience using another survey. One main result was that the OLS experience served as an effective introduction to the profession for first-year PSTs and fourth- and third-year PSTs reported learning in-depth about online teaching in terms of the planning, teaching, and reflecting cycle. Another main result was that PSTs had mathematics teaching anxiety from "a little" to "a moderate amount" before the OLS and their teaching anxiety did not significantly change during the OLS period of 8 weeks. PSTs experienced highest mathematics teaching anxiety when they were observed and evaluated by supervisors during their teaching. The implications of these findings are discussed for teacher education programs.

6.
Hum Brain Mapp ; 42(12): 3963-3982, 2021 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-34043249

RESUMO

Recognising the identity of voices is a key ingredient of communication. Visual mechanisms support this ability: recognition is better for voices previously learned with their corresponding face (compared to a control condition). This so-called 'face-benefit' is supported by the fusiform face area (FFA), a region sensitive to facial form and identity. Behavioural findings indicate that the face-benefit increases in noisy listening conditions. The neural mechanisms for this increase are unknown. Here, using functional magnetic resonance imaging, we examined responses in face-sensitive regions while participants recognised the identity of auditory-only speakers (previously learned by face) in high (SNR -4 dB) and low (SNR +4 dB) levels of auditory noise. We observed a face-benefit in both noise levels, for most participants (16 of 21). In high-noise, the recognition of face-learned speakers engaged the right posterior superior temporal sulcus motion-sensitive face area (pSTS-mFA), a region implicated in the processing of dynamic facial cues. The face-benefit in high-noise also correlated positively with increased functional connectivity between this region and voice-sensitive regions in the temporal lobe in the group of 16 participants with a behavioural face-benefit. In low-noise, the face-benefit was robustly associated with increased responses in the FFA and to a lesser extent the right pSTS-mFA. The findings highlight the remarkably adaptive nature of the visual network supporting voice-identity recognition in auditory-only listening conditions.


Assuntos
Percepção Auditiva/fisiologia , Conectoma , Reconhecimento Facial/fisiologia , Reconhecimento Psicológico/fisiologia , Lobo Temporal/fisiologia , Voz , Adulto , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Ruído , Lobo Temporal/diagnóstico por imagem , Adulto Jovem
7.
Int J Mol Sci ; 22(3)2021 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-33498785

RESUMO

: Phosphorous, in the form of phosphate, is a key element in the nutrition of all living beings. In nature, it is present in the form of phosphate salts, organophosphates, and phosphonates. Bacteria transport inorganic phosphate by the high affinity phosphate transport system PstSCAB, and the low affinity PitH transporters. The PstSCAB system consists of four components. PstS is the phosphate binding protein and discriminates between arsenate and phosphate. In the Streptomyces species, the PstS protein, attached to the outer side of the cell membrane, is glycosylated and released as a soluble protein that lacks its phosphate binding ability. Transport of phosphate by the PstSCAB system is drastically regulated by the inorganic phosphate concentration and mediated by binding of phosphorylated PhoP to the promoter of the PstSCAB operon. In Mycobacterium smegmatis, an additional high affinity transport system, PhnCDE, is also under PhoP regulation. Additionally, Streptomyces have a duplicated low affinity phosphate transport system encoded by the pitH1-pitH2 genes. In this system phosphate is transported as a metal-phosphate complex in simport with protons. Expression of pitH2, but not that of pitH1 in Streptomyces coelicolor, is regulated by PhoP. Interestingly, in many Streptomyces species, three gene clusters pitH1-pstSCAB-ppk (for a polyphosphate kinase), are linked in a supercluster formed by nine genes related to phosphate metabolism. Glycerol-3-phosphate may be transported by the actinobacteria Corynebacterium glutamicum that contains a ugp gene cluster for glycerol-3-P uptake, but the ugp cluster is not present in Streptomyces genomes. Sugar phosphates and nucleotides are used as phosphate source by the Streptomyces species, but there is no evidence of the uhp gene involved in the transport of sugar phosphates. Sugar phosphates and nucleotides are dephosphorylated by extracellular phosphatases and nucleotidases. An isolated uhpT gene for a hexose phosphate antiporter is present in several pathogenic corynebacteria, such as Corynebacterium diphtheriae, but not in non-pathogenic ones. Phosphonates are molecules that contains phosphate linked covalently to a carbon atom through a very stable C-P bond. Their utilization requires the phnCDE genes for phosphonates/phosphate transport and genes for degradation, including those for the subunits of the C-P lyase. Strains of the Arthrobacter and Streptomyces genera were reported to degrade simple phosphonates, but bioinformatic analysis reveals that whole sets of genes for putative phosphonate degradation are present only in three Arthrobacter species and a few Streptomyces species. Genes encoding the C-P lyase subunits occur in several Streptomyces species associated with plant roots or with mangroves, but not in the laboratory model Streptomyces species; however, the phnCDE genes that encode phosphonates/phosphate transport systems are frequent in Streptomyces species, suggesting that these genes, in the absence of C-P lyase genes, might be used as surrogate phosphate transporters. In summary, Streptomyces and related actinobacteria seem to be less versatile in phosphate transport systems than Enterobacteria.


Assuntos
Actinobacteria/metabolismo , Proteínas de Bactérias/metabolismo , Monoéster Fosfórico Hidrolases/metabolismo , Actinobacteria/genética , Arseniatos/metabolismo , Proteínas de Bactérias/genética , Transporte Biológico , Regulação Bacteriana da Expressão Gênica , Glicosilação , Organofosfonatos/metabolismo , Proteínas de Transporte de Fosfato/genética , Proteínas de Transporte de Fosfato/metabolismo , Fosfatos/metabolismo , Regiões Promotoras Genéticas , Transdução de Sinais , Streptomyces/genética , Streptomyces/metabolismo , Ácidos Teicoicos/metabolismo
8.
Shokuhin Eiseigaku Zasshi ; 62(3): 85-93, 2021.
Artigo em Japonês | MEDLINE | ID: mdl-34219101

RESUMO

The mouse bioassay (MBA) for paralytic shellfish toxins (PSTs) in bivalves has been used as an official method in Japan. It is necessary to develop an alternative method to animal experiments in PSTs assay because 3Rs (Replacement, Reduction, and Refinement) of animal experiments are required from the animal welfare point of view. Various methods such as HPLC-FL, receptor binding assay, LC-MS/MS and ELISA have been established to detect PSTs without performing animal experiments. The present study was undertaken to develop a screening method using oligonucleotide lateral flow immunoassay (OLFIA) for detecting PSTs in bivalves. The screening level was defined as positive at 2 MU/g of MBA that is the half regulation limit of PSTs monitoring in Japan. All 20 positive (equal to or more than 2 MU/g) samples judged from MBA showed a positive reaction in the OLFIA. No positive samples resulted in a false negative reaction. The OLFIA exhibited high accuracy at 2 MU/g of screening criteria. The authors demonstrated here that the OLFIA can be useful for rapid detection of PSTs in bivalves.


Assuntos
Bivalves , Intoxicação por Frutos do Mar , Animais , Cromatografia Líquida , Imunoensaio , Japão , Toxinas Marinhas/análise , Camundongos , Oligonucleotídeos , Saxitoxina/análise , Frutos do Mar/análise , Espectrometria de Massas em Tandem
9.
J Infect Dis ; 222(7): 1204-1212, 2020 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-32324853

RESUMO

Acinetobacter baumannii is a successful pathogen responsible for infections with high mortality rate. During the course of infection it can be found in microaerobic environments, which influences virulence factor expression. From a previous transcriptomic analysis of A. baumannii ATCC 17978 under microaerobiosis, we know the gene pstS is overexpressed under microaerobiosis. Here, we studied its role in A. baumannii virulence. pstS loss significantly decreased bacterial adherence and invasion into A549 cells and increased A549 cell viability. pstS loss also reduced motility and biofilm-forming ability of A. baumannii. In a peritoneal sepsis murine model, the minimum lethal dose required by A. baumannii ATCC 17978 ΔpstS was lower compared to the wild type (4.3 vs 3.2 log colony forming units/mL, respectively), and the bacterial burden in tissues and fluids was lower. Thus, the loss of the phosphate sensor PstS produced a decrease in A. baumannii pathogenesis, supporting its role as a virulence factor.


Assuntos
Acinetobacter baumannii/genética , Acinetobacter baumannii/patogenicidade , Proteínas de Bactérias/genética , Proteínas de Ligação a Fosfato/genética , Células A549 , Infecções por Acinetobacter/microbiologia , Acinetobacter baumannii/crescimento & desenvolvimento , Aerobiose , Animais , Aderência Bacteriana/genética , Biofilmes , Morte Celular , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Feminino , Regulação Bacteriana da Expressão Gênica , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Oxigênio/farmacologia , Peritonite/microbiologia , Virulência/genética , Fatores de Virulência/genética
10.
Emerg Infect Dis ; 26(9): 2247-2250, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32818423

RESUMO

Rates of vancomycin-resistant enterococci bloodstream infections have remained relatively low in Canada. We recently observed an increase of 113% in these infections rates, which coincided with emergence of Enterococcus faecium pstS-null sequence type 1478. The proportion of this sequence type increased from 2.7% to 38.7% for all tested isolates from 2013-2018.


Assuntos
Enterococcus faecium , Infecções por Bactérias Gram-Positivas , Enterococos Resistentes à Vancomicina , Antibacterianos/farmacologia , Canadá/epidemiologia , Células Clonais , Enterococcus faecium/genética , Infecções por Bactérias Gram-Positivas/epidemiologia , Humanos , Testes de Sensibilidade Microbiana , Vancomicina/farmacologia , Enterococos Resistentes à Vancomicina/genética
11.
Eur J Clin Microbiol Infect Dis ; 39(7): 1349-1356, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32128641

RESUMO

Although multilocus sequence typing (MLST) has been used to study molecular epidemiology and to explore the population structure of Enterococcus faecium, vancomycin-resistant E. faecium (VREF) strains lacking the pstS gene that were non-typable using conventional MLST methods were reported recently. We found nationwide emergence of VREF isolates lacking pstS in Korea and hereby report the molecular characteristics of these isolates. Forty-six VREF isolates lacking the pstS gene were identified among 300 VREF rectal isolates collected from hospitalized patients between 2014 and 2015. MLST was performed and clonal relatedness was determined by pulsed-field gel electrophoresis (PFGE). Four VREF ST1421 isolates were whole-genome sequenced. Among the VREF rectal isolates lacking pstS, 98% were classified as ST1421, which has identical allelic profiles to ST17 for all housekeeping genes except pstS. PFGE pattern analyses revealed 32 pulsotypes. All isolates harbored Tn1546 components with various transposase and insertion sequences. The whole-genome sequencing of four VREF ST1421 isolates showed that the pstS gene region was deleted at various locations with considerable inversion. The pstS gene was also depleted in 12.1% of 33 VREF clinical isolates in 2006-2007 and in 11.8% of 59 clinical isolates in 2012-2013. VREF ST1421 strains lacking the pstS gene have emerged in Korea. The emergence and spread of pstS-deleted VREF strains pose a serious challenge for epidemiological investigation. Alternative molecular typing methods to MLST will be increasingly necessary.


Assuntos
Proteínas de Bactérias/genética , Enterococcus faecium/genética , Infecções por Bactérias Gram-Positivas/microbiologia , Proteínas de Ligação a Fosfato/genética , Enterococos Resistentes à Vancomicina/genética , Alelos , Elementos de DNA Transponíveis , Enterococcus faecium/classificação , Enterococcus faecium/isolamento & purificação , Deleção de Genes , Genes Essenciais/genética , Genoma Bacteriano/genética , Infecções por Bactérias Gram-Positivas/epidemiologia , Humanos , Tipagem de Sequências Multilocus , Filogenia , Prevalência , República da Coreia/epidemiologia , Enterococos Resistentes à Vancomicina/efeitos dos fármacos , Enterococos Resistentes à Vancomicina/isolamento & purificação
12.
Lett Appl Microbiol ; 70(1): 2-12, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31518435

RESUMO

Dental caries, caused by Streptococcus mutans, is a common infection. Caries vaccine has been under investigation for the last 40 years. Many in vitro and in vivo studies and some human clinical trials have determined many pertinent aspects regarding vaccine development. The virulence determinants of Strep. mutans, such as Ag I/II, responsible for adherence to surfaces, glucosyltransferase, responsible for the production of glucan, and the glucan-binding protein, responsible for the attachment of glucan to surfaces, have been known to elicit an antigen-specific immune response. It is also known that more than one antigen or a functional part of the genome responsible for these virulence determinants provide a better host response compared with the monogenic vaccine or complete genome of a specific antigen. To enhance the host response, the use of adjuvants has been studied and the routes of antigen administration have been investigated. In recent years, some promising vaccines such as pGJA-P/VAX, LT derivative/Pi39-512 , KFD2-rPAc and SBR/GBR-CMV-nirB have been developed and tested in animals. New virulence targets need to be explored. Multicentre collaborative studies and human clinical trials are required and some interest from funders and public health experts should be generated to overcome this hurdle. SIGNIFICANCE AND IMPACT OF THE STUDY: Dental caries is an irreversible, multifactorial opportunistic infection. The treatment is costly, making it a public health problem. Despite many years of promising laboratory research, animal studies and clinical trials, there is no commercially available vaccine today. The research objectives have become more refined from lessons learnt over the years. Multigenic DNA/recombinant vaccines, using the best proved adjuvants with a delivery system for the nasal or sublingual route, should be developed and researched with multicentre collaborative efforts. In addition, new vaccine targets can be identified. To overcome the economic hurdle, funders and public health interest should be stimulated.


Assuntos
Vacinas Bacterianas/imunologia , Cárie Dentária/prevenção & controle , Streptococcus mutans/imunologia , Animais , Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/genética , Cárie Dentária/microbiologia , Humanos , Streptococcus mutans/genética , Streptococcus mutans/fisiologia , Vacinas de DNA/administração & dosagem , Vacinas de DNA/genética , Vacinas de DNA/imunologia
13.
Neurosurg Focus ; 48(2): E10, 2020 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-32006947

RESUMO

The object of this study was to extensively characterize a region of periventricular nodular heterotopia (PVNH) in an epilepsy patient to reveal its possible neurocognitive functional role(s). The authors used 3-T MRI approaches to exhaustively characterize a single, right hemisphere heterotopion in a high-functioning adult male with medically responsive epilepsy, which had manifested during late adolescence. The heterotopion proved to be spectroscopically consistent with a cortical-like composition and was interconnected with nearby ipsilateral cortical fundi, as revealed by fiber tractography (diffusion-weighted imaging) and resting-state functional connectivity MRI (rsfMRI). Moreover, the region of PVNH demonstrated two novel characterizations for a heterotopion. First, functional MRI (fMRI), as distinct from rsfMRI, showed that the heterotopion was significantly modulated while the patient watched animated video scenes of biological motion (i.e., cartoons). Second, rsfMRI, which demonstrated correlated brain activity during a task-negative state, uniquely showed directionality within an interconnected network, receiving positive path effects from patent cortical and cerebellar foci while outputting only negative path effects to specific brain foci.These findings are addressed in the context of the impact on noninvasive presurgical brain mapping strategies for adult and pediatric patient workups, as well as the impact of this study on an understanding of the functional cortical architecture underlying cognition from a neurodiversity and evolutionary perspective.


Assuntos
Mapeamento Encefálico/métodos , Epilepsia/diagnóstico por imagem , Imageamento por Ressonância Magnética/métodos , Heterotopia Nodular Periventricular/diagnóstico por imagem , Descanso/fisiologia , Convulsões/diagnóstico por imagem , Epilepsia/fisiopatologia , Humanos , Masculino , Heterotopia Nodular Periventricular/fisiopatologia , Cuidados Pré-Operatórios/métodos , Convulsões/fisiopatologia , Adulto Jovem
14.
J Environ Sci (China) ; 92: 129-140, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32430116

RESUMO

Simultaneous overabundance and scarcity of inorganic phosphate (Pi) is a critical issue driving the development of innovative water/wastewater treatment technologies that not only facilitate Pi removal to prevent eutrophication, but also recover Pi for agricultural reuse. Here, a cell-surface expressed high-affinity phosphate binding protein (PstS) system was developed, and its Pi capture and release potential was evaluated. E. coli was genetically modified to express PstS on its outer membrane using the ice nucleation protein (INP) as an anchoring motif. Verification of protein expression and localization were performed utilizing SDS-polyacrylamide gel electrophoresis (SDS-PAGE), western blot, and outer membrane separation analyses. Cell surface characterization was investigated through acid-base titration, X-ray photoelectron spectroscopy (XPS), and Fourier transform infrared spectroscopy (FTIR). These tests provided information on the macromolecular structure and composition of the bacteria surface as well as the proton-exchange properties of the surface functional groups (i.e., pKa values). Phosphate desorption and adsorption batch experiments were conducted to evaluate the effects of temperature, pH, and ionic strength on phosphate capture and release. The PstS surface-displayed cells demonstrated greater potential to release and capture phosphate compared to non-modified cells. Higher temperatures up to 40°C, basic pH conditions (pH = 10.5), and higher ionic strength up to 1.0 mol/L KCl promoted 20%-50% higher phosphate release.


Assuntos
Fosfatos , Fósforo , Adsorção , Proteínas de Transporte , Escherichia coli , Proteínas de Ligação a Fosfato
15.
Neuroimage ; 198: 296-302, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31100434

RESUMO

Recent behavioural evidence shows that visual displays of two individuals interacting are not simply encoded as separate individuals, but as an interactive unit that is 'more than the sum of its parts'. Recent functional magnetic resonance imaging (fMRI) evidence shows the importance of the posterior superior temporal sulcus (pSTS) in processing human social interactions, and suggests that it may represent human-object interactions as qualitatively 'greater' than the average of their constituent parts. The current study aimed to investigate whether the pSTS or other posterior temporal lobe region(s): 1) Demonstrated evidence of a dyadic information effect - that is, qualitatively different responses to an interacting dyad than to averaged responses of the same two interactors, presented in isolation, and; 2) Significantly differentiated between different types of social interactions. Multivoxel pattern analysis was performed in which a classifier was trained to differentiate between qualitatively different types of dyadic interactions. Above-chance classification of interactions was observed in 'interaction selective' pSTS-I and extrastriate body area (EBA), but not in other regions of interest (i.e. face-selective STS and mentalizing-selective temporo-parietal junction). A dyadic information effect was not observed in the pSTS-I, but instead was shown in the EBA; that is, classification of dyadic interactions did not fully generalise to averaged responses to the isolated interactors, indicating that dyadic representations in the EBA contain unique information that cannot be recovered from the interactors presented in isolation. These findings complement previous observations for congruent grouping of human bodies and objects in the broader lateral occipital temporal cortex area.


Assuntos
Relações Interpessoais , Lobo Occipital/fisiologia , Percepção Social , Lobo Temporal/fisiologia , Adolescente , Adulto , Mapeamento Encefálico , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Reconhecimento Visual de Modelos/fisiologia , Máquina de Vetores de Suporte , Adulto Jovem
16.
Neuroimage ; 184: 90-100, 2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30217542

RESUMO

What is a face? Intuition, along with abundant behavioral and neural evidence, indicates that internal features (e.g., eyes, nose, mouth) are critical for face recognition, yet some behavioral and neural findings suggest that external features (e.g., hair, head outline, neck and shoulders) may likewise be processed as a face. Here we directly test this hypothesis by investigating how external (and internal) features are represented in the brain. Using fMRI, we found highly selective responses to external features (relative to objects and scenes) within the face processing system in particular, rivaling that observed for internal features. We then further asked how external and internal features are represented in regions of the cortical face processing system, and found a similar division of labor for both kinds of features, with the occipital face area and posterior superior temporal sulcus representing the parts of both internal and external features, and the fusiform face area representing the coherent arrangement of both internal and external features. Taken together, these results provide strong neural evidence that a "face" is composed of both internal and external features.


Assuntos
Córtex Cerebral/fisiologia , Reconhecimento Facial/fisiologia , Adulto , Mapeamento Encefálico , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Lobo Occipital/fisiologia , Lobo Temporal/fisiologia , Adulto Jovem
17.
Microb Cell Fact ; 18(1): 11, 2019 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-30660186

RESUMO

BACKGROUND: Pichia pastoris (syn. Komagataella phaffii) is one of the most highly utilized eukaryotic expression systems for the production of heterologous glycoproteins, being able to perform both N- and O-mannosylation. In this study, we present the expression in P. pastoris of an O-mannosylated recombinant version of the 38 kDa glycolipoprotein PstS-1 from Mycobacterium tuberculosis (Mtb), that is similar in primary structure to the native secreted protein. RESULTS: The recombinant PstS-1 (rPstS-1) was produced without the native lipidation signal. Glycoprotein expression was under the control of the methanol-inducible promoter pAOX1, with secretion being directed by the α-mating factor secretion signal. Production of rPstS-1 was carried out in baffled shake flasks (BSFs) and controlled bioreactors. A production up to ~ 46 mg/L of the recombinant protein was achieved in both the BSFs and the bioreactors. The recombinant protein was recovered from the supernatant and purified in three steps, achieving a preparation with 98% electrophoretic purity. The primary and secondary structures of the recombinant protein were characterized, as well as its O-mannosylation pattern. Furthermore, a cross-reactivity analysis using serum antibodies from patients with active tuberculosis demonstrated recognition of the recombinant glycoprotein, indirectly indicating the similarity between the recombinant PstS-1 and the native protein from Mtb. CONCLUSIONS: rPstS-1 (98.9% sequence identity, O-mannosylated, and without tags) was produced and secreted by P. pastoris, demonstrating that this yeast is a useful cell factory that could also be used to produce other glycosylated Mtb antigens. The rPstS-1 could be used as a tool for studying the role of this molecule during Mtb infection, and to develop and improve vaccines or kits based on the recombinant protein for serodiagnosis.


Assuntos
Transportadores de Cassetes de Ligação de ATP/metabolismo , Proteínas de Bactérias/metabolismo , Mycobacterium tuberculosis/metabolismo , Pichia/metabolismo , Transportadores de Cassetes de Ligação de ATP/química , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/imunologia , Aldeído Oxidase/genética , Anticorpos Antibacterianos/imunologia , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Reatores Biológicos , Dicroísmo Circular , Eletroforese em Gel de Poliacrilamida , Glicosilação , Humanos , Pichia/crescimento & desenvolvimento , Plasmídeos/metabolismo , Regiões Promotoras Genéticas , Estrutura Secundária de Proteína , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
18.
J Phycol ; 55(4): 752-761, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-30929262

RESUMO

The marine diazotroph Crocosphaera watsonii provides fixed carbon (C) and nitrogen (N) to open-ocean regimes, where nutrient deficiency controls productivity. The growth of Crocosphaera can be limited by low phosphorus (P) concentrations in these oligotrophic environments. Biomarkers such as the high-affinity ABC transporter phosphate-binding gene, pstS, are commonly used to monitor when such organisms are under P stress; however, transcriptional regulation of these markers is often complex and not well-understood. In this study, we interrogated changes in pstS transcript levels in C. watsonii cells under P starvation, and in response to added dissolved inorganic phosphorus (DIP), dissolved organic phosphorus (DOP), and changing light levels. We observed elevated relative pstS transcript levels in C. watsonii WH8501 at DIP concentrations below 60 and above 20 nmol · L-1 . Transcript levels were suppressed by both inorganic and bioavailable organic phosphorus; however, the P stress response was more sensitive to DIP than DOP sources. Increasing light intensity resulted in increased relative pstS transcript abundances independently of low external P, and seemed to exacerbate the physiological effects of P stress. The variable response to different P compounds and rapid and transient influence of high light on pstS transcript abundances suggests that pstS is an indicator of internal P status in Crocosphaera.


Assuntos
Cianobactérias , Fósforo , Fixação de Nitrogênio , Proteínas de Transporte de Fosfato
19.
Hum Brain Mapp ; 39(12): 5112-5125, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30273447

RESUMO

The posterior superior temporal sulcus (pSTS) plays an important role in biological motion perception but is also thought to be essential for speech and facial processing. However, although there are many previous investigations of distinct functional modules within the pSTS, the functional organization of the pSTS in its full functional heterogeneity has not yet been established. Here we applied a connectivity-based parcellation strategy to delineate the human pSTS subregions based on distinct anatomical connectivity profiles and divided it into rostral and caudal subregions using diffusion tensor imaging. Subsequent multimodal connection pattern analyses revealed distinct subregional connectivity profiles. From this we inferred that the two subregions are involved in distinct functional circuits, the language processing loop and the cognition attention network. These results indicate a convergent functional architecture of the pSTS that can be revealed based on different types of connectivity and is reflected in different functions and interactions. In addition, when the subregions were performing their processing in the different functional circuits, we found asymmetry in the bilateral pSTS. Our findings may improve the understanding of the functional organization of the pSTS and provide new insights into its interactions and integration of information at the subregional level.


Assuntos
Encéfalo , Conectoma/métodos , Imagem de Tensor de Difusão/métodos , Rede Nervosa , Lobo Temporal , Adulto , Encéfalo/anatomia & histologia , Encéfalo/diagnóstico por imagem , Encéfalo/fisiologia , Feminino , Humanos , Masculino , Rede Nervosa/anatomia & histologia , Rede Nervosa/diagnóstico por imagem , Rede Nervosa/fisiologia , Lobo Temporal/anatomia & histologia , Lobo Temporal/diagnóstico por imagem , Lobo Temporal/fisiologia , Adulto Jovem
20.
Psychol Med ; 48(11): 1862-1871, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29173243

RESUMO

BACKGROUND: Recognition of communicative interactions is a complex social cognitive ability which is associated with a specific neural activity in healthy individuals. However, neural correlates of communicative interaction processing from whole-body motion have not been known in patients with schizophrenia (SCZ). Therefore, the current study aims to examine the neural activity associated with recognition of communicative interactions in SCZ by using displays of the dyadic interactions downgraded to minimalistic point-light presentations. METHODS: Twenty-six healthy controls (HC) and 25 SCZ were asked to judge whether two agents presented only by point-light displays were communicating or acting independently. Task-related activity and functional connectivity of brain structures were examined with General Linear Model and Generalized Psychophysiological Interaction approach, respectively. RESULTS: HC were significantly more efficient in recognizing each type of action than SCZ. At the neural level, the activity of the right posterior superior temporal sulcus (pSTS) was observed to be higher in HC compared with SCZ for communicative v. individual action processing. Importantly, increased connectivity of the right pSTS with structures associated with mentalizing (left pSTS) and mirroring networks (left frontal areas) was observed in HC, but not in SCZ, during the presentation of social interactions. CONCLUSION: Under-recruitment of the right pSTS, a structure known to have a pivotal role in social processing, may also be of importance for higher-order social cognitive deficits in SCZ. Furthermore, decreased task-related connectivity of the right pSTS may result in reduced use of additional sources of information (for instance motor resonance signals) during social cognitive processing in schizophrenia.


Assuntos
Córtex Cerebral/fisiopatologia , Conectoma/métodos , Relações Interpessoais , Percepção de Movimento/fisiologia , Reconhecimento Visual de Modelos/fisiologia , Esquizofrenia/fisiopatologia , Percepção Social , Adulto , Córtex Cerebral/diagnóstico por imagem , Comunicação , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Adulto Jovem
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