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1.
Nature ; 628(8007): 424-432, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38509359

RESUMO

Fusobacterium nucleatum (Fn), a bacterium present in the human oral cavity and rarely found in the lower gastrointestinal tract of healthy individuals1, is enriched in human colorectal cancer (CRC) tumours2-5. High intratumoural Fn loads are associated with recurrence, metastases and poorer patient prognosis5-8. Here, to delineate Fn genetic factors facilitating tumour colonization, we generated closed genomes for 135 Fn strains; 80 oral strains from individuals without cancer and 55 unique cancer strains cultured from tumours from 51 patients with CRC. Pangenomic analyses identified 483 CRC-enriched genetic factors. Tumour-isolated strains predominantly belong to Fn subspecies animalis (Fna). However, genomic analyses reveal that Fna, considered a single subspecies, is instead composed of two distinct clades (Fna C1 and Fna C2). Of these, only Fna C2 dominates the CRC tumour niche. Inter-Fna analyses identified 195 Fna C2-associated genetic factors consistent with increased metabolic potential and colonization of the gastrointestinal tract. In support of this, Fna C2-treated mice had an increased number of intestinal adenomas and altered metabolites. Microbiome analysis of human tumour tissue from 116 patients with CRC demonstrated Fna C2 enrichment. Comparison of 62 paired specimens showed that only Fna C2 is tumour enriched compared to normal adjacent tissue. This was further supported by metagenomic analysis of stool samples from 627 patients with CRC and 619 healthy individuals. Collectively, our results identify the Fna clade bifurcation, show that specifically Fna C2 drives the reported Fn enrichment in human CRC and reveal the genetic underpinnings of pathoadaptation of Fna C2 to the CRC niche.


Assuntos
Neoplasias Colorretais , Fusobacterium nucleatum , Animais , Humanos , Camundongos , Adenoma/microbiologia , Estudos de Casos e Controles , Neoplasias Colorretais/microbiologia , Neoplasias Colorretais/patologia , Fezes/microbiologia , Fusobacterium nucleatum/classificação , Fusobacterium nucleatum/genética , Fusobacterium nucleatum/isolamento & purificação , Fusobacterium nucleatum/patogenicidade , Trato Gastrointestinal/metabolismo , Trato Gastrointestinal/microbiologia , Genoma Bacteriano/genética , Boca/microbiologia , Feminino
2.
J Virol ; 97(12): e0173723, 2023 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-38051260

RESUMO

IMPORTANCE: Viruses are constantly evolving to promote propagation in the host. Here, we show that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) utilizes host RAD51 for replication. Silencing of RAD51 impaired SARS-CoV-2 propagation. Viral RNA colocalized with RAD51 in the cytoplasm of SARS-CoV-2-infected cells, suggesting that both viral RNA and RAD51 may form a replication complex. We, therefore, evaluated RAD51 inhibitors as possible therapeutic agents against SARS-CoV-2. Indeed, RAD51 inhibitors exerted antiviral activities against not only Wuhan but also variants of SARS-CoV-2. Molecular docking model shows that RAD51 inhibitors impede SARS-CoV-2 propagation by interfering with dimerization of RAD51. These data suggest that RAD51 may represent a novel host-based drug target for coronavirus disease 2019 treatment.


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , COVID-19/metabolismo , COVID-19/virologia , Simulação de Acoplamento Molecular , Rad51 Recombinase/antagonistas & inibidores , Rad51 Recombinase/metabolismo , RNA Viral , SARS-CoV-2/fisiologia , Interações Hospedeiro-Patógeno
3.
BMC Gastroenterol ; 24(1): 80, 2024 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-38388860

RESUMO

OBJECTIVES: Poorly visualized images that appear during small bowel capsule endoscopy (SBCE) can confuse the interpretation of small bowel lesions and increase the physician's workload. Using a validated artificial intelligence (AI) algorithm that can evaluate the mucosal visualization, we aimed to assess whether SBCE reading after the removal of poorly visualized images could affect the diagnosis of SBCE. METHODS: A study was conducted to analyze 90 SBCE cases in which a small bowel examination was completed. Two experienced endoscopists alternately performed two types of readings. They used the AI algorithm to remove poorly visualized images for the frame reduction reading (AI user group) and conducted whole frame reading without AI (AI non-user group) for the same patient. A poorly visualized image was defined as an image with < 50% mucosal visualization. The study outcomes were diagnostic concordance and reading time between the two groups. The SBCE diagnosis was classified as Crohn's disease, bleeding, polyp, angiodysplasia, and nonspecific finding. RESULTS: The final SBCE diagnoses between the two groups showed statistically significant diagnostic concordance (k = 0.954, p < 0.001). The mean number of lesion images was 3008.5 ± 9964.9 in the AI non-user group and 1401.7 ± 4811.3 in the AI user group. There were no cases in which lesions were completely removed. Compared with the AI non-user group (120.9 min), the reading time was reduced by 35.6% in the AI user group (77.9 min). CONCLUSIONS: SBCE reading after reducing poorly visualized frames using the AI algorithm did not have a negative effect on the final diagnosis. SBCE reading method integrated with frame reduction and mucosal visualization evaluation will help improve AI-assisted SBCE interpretation.


Assuntos
Endoscopia por Cápsula , Doenças do Colo , Doença de Crohn , Humanos , Inteligência Artificial , Intestino Delgado/diagnóstico por imagem , Intestino Delgado/patologia , Doença de Crohn/diagnóstico por imagem , Doença de Crohn/cirurgia , Estudos Retrospectivos
4.
J Comput Aided Mol Des ; 37(9): 453-461, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37452977

RESUMO

African swine fever virus (ASFV), an extremely contagious virus with high mortality rates, causes severe hemorrhagic viral disease in both domestic and wild pigs. Fortunately, ASFV cannot be transmitted from pigs to humans. However, ongoing ASFV outbreaks could have severe economic consequences for global food security. Although ASFV was discovered several years ago, no vaccines or treatments are commercially available yet; therefore, the identification of new anti-ASFV drugs is urgently warranted. Using molecular docking and machine learning, we have previously identified pentagastrin, cangrelor, and fostamatinib as potential antiviral drugs against ASFV. Here, using machine learning combined with docking simulations, we identified natural products with a high affinity for AsfvPolX proteins. We selected five natural products (NPs) that are located close in chemical space to the six known natural flavonoids that possess anti-ASFV activity. Polygalic acid markedly reduced AsfvPolX polymerase activity in a dose-dependent manner. We propose an efficient protocol for identifying NPs as potential antiviral drugs by identifying chemical spaces containing high-affinity binders against ASFV in NP databases.


Assuntos
Vírus da Febre Suína Africana , Febre Suína Africana , Humanos , Suínos , Animais , Febre Suína Africana/prevenção & controle , Simulação de Acoplamento Molecular , Proteínas Virais , Antivirais/farmacologia , Técnicas In Vitro , Aprendizado de Máquina
5.
Dig Dis Sci ; 68(3): 867-876, 2023 03.
Artigo em Inglês | MEDLINE | ID: mdl-35781655

RESUMO

BACKGROUND: Efficient bowel preparation is essential for preventing colorectal cancer by improving endoscopic adenoma detection. Tablet for bowel preparation containing sulfate salts, OSTs (oral sulfate tablets), has been developed and it is gaining more popularity. However, its efficacy compared to standard preparation agent, PEG-AA (polyethylene glycol), has not been well discovered. We assessed the efficacy of PEG and OSTs using a real-time clinical data warehouse (CDW) model. METHODS: We performed a propensity score-matched (PSM) analysis of consecutive adult patients undergoing colonoscopy who received PEG-AA or OSTs prior to colonoscopy at a tertiary academic hospital. The endoscopic records of 992 adult patients were retrospectively analyzed. The clinical data warehouse collected data including bowel preparation, insertion time, observation time, and the detection of polyps and adenomas. Multivariate regression analysis was performed to reveal the factors associated with endoscopic outcomes. RESULTS: Among 992 patients included in the study, 770 and 222 patients received PEG-AA and OSTs, respectively. Among the propensity score-matched population (n = 1897), OSTs resulted in better bowel cleansing quality (8.16 vs 7.84, p = 0.014) and a higher adenoma detection rate (38.6% vs 27.1%, p = 0.003). Using PEG-AA, older age, inadequate bowel preparation (BBPS score < 6) and endoscopy by fellows were found to be factors associated with poor adenoma detection. In the elderly over 65 years of age, a significant difference in cleansing quality between the two groups (7.21 vs 8.19, p < 0.001) was found, but its impact on ADR was not prominent (49.5% vs 45.4%, p = 0.653). CONCLUSIONS: OSTs with simethicone achieved better endoscopic cleanliness, improving adenoma detection rate compared to the conventional PEG-AA protocol. The synergistic effect of both the convenience of taking tablets and the reduction of intraluminal bubble by adjunctive simethicone improves the clinical efficacy of colonoscopy.


Assuntos
Adenoma , Polietilenoglicóis , Adulto , Humanos , Idoso , Polietilenoglicóis/efeitos adversos , Simeticone , Catárticos/efeitos adversos , Sulfatos , Pontuação de Propensão , Estudos Retrospectivos , Colonoscopia/métodos , Adenoma/diagnóstico , Adenoma/induzido quimicamente , Comprimidos
6.
Mol Ther ; 30(5): 1994-2004, 2022 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-35007757

RESUMO

Adeno-associated virus (AAV)-mediated gene delivery holds great promise for gene therapy. However, the non-invasive delivery of AAV for lung tissues has not been adequately established. Here, we revealed that the intratracheal administration of an appropriate amount of AAV2/8 predominantly targets lung tissue. AAV-mediated gene delivery that we used in this study induced the expression of the desired protein in lung parenchymal cells, including alveolar type II cells. We harnessed the technique to develop severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-susceptible mice. Three kinds of immune function-relevant gene knockout (KO) mice were transduced with AAV encoding human angiotensin-converting enzyme 2 (hACE2) and then injected with SARS-CoV-2. Among these mice, type I interferon receptor (IFNAR) KO mice showed increased viral titer in the lungs compared to that in the other KO mice. Moreover, nucleocapsid protein of SARS-CoV-2 and multiple lesions in the trachea and lung were observed in AAV-hACE2-transduced, SARS-CoV-2-infected IFNAR KO mice, indicating the involvement of type I interferon signaling in the protection of SARS-CoV-2. In this study, we demonstrate the ease and rapidness of the intratracheal administration of AAV for targeting lung tissue in mice, and this can be used to study diverse pulmonary diseases.


Assuntos
COVID-19 , SARS-CoV-2 , Animais , COVID-19/terapia , Dependovirus/genética , Modelos Animais de Doenças , Suscetibilidade a Doenças , Pulmão/patologia , Camundongos , Camundongos Transgênicos , SARS-CoV-2/genética
7.
BMC Med Educ ; 23(1): 12, 2023 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-36624494

RESUMO

BACKGROUND: Reflective writing (RW) allows physicians to step back, review their thoughts, goals and actions and recognise how their perspectives, motives and emotions impact their conduct. RW also helps physicians consolidate their learning and boosts their professional and personal development. In the absence of a consistent approach and amidst growing threats to RW's place in medical training, a review of theories of RW in medical education and a review to map regnant practices, programs and assessment methods are proposed. METHODS: A Systematic Evidence-Based Approach guided Systematic Scoping Review (SSR in SEBA) was adopted to guide and structure the two concurrent reviews. Independent searches were carried out on publications featured between 1st January 2000 and 30th June 2022 in PubMed, Embase, PsychINFO, CINAHL, ERIC, ASSIA, Scopus, Google Scholar, OpenGrey, GreyLit and ProQuest. The Split Approach saw the included articles analysed separately using thematic and content analysis. Like pieces of a jigsaw puzzle, the Jigsaw Perspective combined the themes and categories identified from both reviews. The Funnelling Process saw the themes/categories created compared with the tabulated summaries. The final domains which emerged structured the discussion that followed. RESULTS: A total of 33,076 abstracts were reviewed, 1826 full-text articles were appraised and 199 articles were included and analysed. The domains identified were theories and models, current methods, benefits and shortcomings, and recommendations. CONCLUSIONS: This SSR in SEBA suggests that a structured approach to RW shapes the physician's belief system, guides their practice and nurtures their professional identity formation. In advancing a theoretical concept of RW, this SSR in SEBA proffers new insight into the process of RW, and the need for longitudinal, personalised feedback and support.


Assuntos
Educação Médica , Médicos , Humanos , Currículo , Aprendizagem , Redação
8.
Biochem Biophys Res Commun ; 595: 14-21, 2022 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-35093635

RESUMO

Organoid cryopreservation method is one of key step in the organoid culture. We aimed to establish a simple and efficient cryopreservation method for mouse small intestinal organoids (MIOs) and colon organoids (MCOs) using various concentrations of cryoprotectant. Based on the theoretical simulation, we optimized the dimethyl sulfoxide (DMSO) concentration by pretreating the organoids with 5, 7.5, and 10% DMSO for 30 min at 4 °C to allow penetration into the organoids and evaluated their viability, proliferation, and function after cryopreservation. Gene expression in the MIOs and staining of lineage markers were examined real-time PCR. The organoids in the DMSO-treated groups as well as the control, expressed ChrgA, Ecad, Muc2, Lyz, villin, and Lgr5, and there are no significant. A forskolin-induced swelling assay for MIOs was performed to confirm normal cystic fibrosis transmembrane conductance regulator (CFTR) activity. Similar forskolin-induced swelling was observed in the DMSO-treated groups and the control. In addition, MCOs were transplanted into mouse colon for confirmation of regeneration therapy efficacy. Thawing organoids were cultured for two and four sequential passages after cryopreservation with 5% DMSO to confirm any changes in the gene expression of lineage markers after subculture. We developed a simple and efficient organoid freezing method using 5% DMSO with low potential toxicity and validated our findings with theoretical simulation.


Assuntos
Colo/metabolismo , Criopreservação/métodos , Intestino Delgado/metabolismo , Organoides/metabolismo , Medicina Regenerativa/métodos , Animais , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Crioprotetores/metabolismo , Crioprotetores/farmacologia , Dimetil Sulfóxido/metabolismo , Dimetil Sulfóxido/farmacologia , Expressão Gênica/efeitos dos fármacos , Camundongos , Organoides/citologia , Organoides/efeitos dos fármacos , Fatores de Tempo
9.
AJR Am J Roentgenol ; 218(3): 506-516, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-34523950

RESUMO

BACKGROUND. Shoulder MRI using standard multiplanar sequences requires long scan times. Accelerated sequences have tradeoffs in noise and resolution. Deep learning-based reconstruction (DLR) may allow reduced scan time with preserved image quality. OBJECTIVE. The purpose of this study was to compare standard shoulder MRI sequences and accelerated sequences without and with DLR in terms of image quality and diagnostic performance. METHODS. This retrospective study included 105 patients (45 men, 60 women; mean age, 57.6 ± 10.9 [SD] years) who underwent a total of 110 3-T shoulder MRI examinations. Examinations included standard sequences (scan time, 9 minutes 23 seconds) and accelerated sequences (3 minutes 5 seconds; 67% reduction), both including fast spin-echo sequences in three planes. Standard sequences were reconstructed using the conventional pipeline; accelerated sequences were reconstructed using both the conventional pipeline and a commercially available DLR pipeline. Two radiologists independently assessed three image sets (standard sequence, accelerated sequence without DLR, and accelerated sequence with DLR) for subjective image quality and artifacts using 4-point scales (4 = highest quality) and identified pathologies of the subscapularis tendon, supraspinatus-infraspinatus tendon, long head of the biceps brachii tendon, and glenoid labrum. Interobserver agreement and agreement between image sets for the evaluated pathologies were assessed using weighted kappa statistics. In 27 patients who underwent arthroscopy, diagnostic performance was calculated using arthroscopic findings as a reference standard. RESULTS. Mean subjective image quality scores for readers 1 and 2 were 10.6 ± 1.2 and 10.5 ± 1.4 for the standard sequence, 8.1 ± 1.3 and 7.2 ± 1.1 for the accelerated sequence without DLR, and 10.7 ± 1.2 and 10.5 ± 1.6 for the accelerated sequence with DLR. Mean artifact scores for readers 1 and 2 were 9.3 ± 1.2 and 10.0 ± 1.0 for the standard sequence, 7.3 ± 1.3 and 9.1 ± 0.8 for the accelerated sequence without DLR, and 9.4 ± 1.2 and 9.8 ± 0.8 for the accelerated sequence with DLR. Interobserver agreement ranged from kappa of 0.813-0.951 except for accelerated sequence without DLR for the supraspinatus-infraspinatus tendon (κ = 0.673). Agreement between image sets ranged from kappa of 0.809-0.957 except for reader 1 for supraspinatus-infraspinatus tendon (κ = 0.663-0.700). Accuracy, sensitivity, and specificity for tears of the four structures were not different (p > .05) among image sets. CONCLUSION. Accelerated sequences with DLR provide 67% scan time reduction with similar subjective image quality, artifacts, and diagnostic performance to standard sequences. CLINICAL IMPACT. Accelerated sequences with DLR may provide an alternative to standard sequences for clinical shoulder MRI.


Assuntos
Aprendizado Profundo , Interpretação de Imagem Assistida por Computador/métodos , Imageamento por Ressonância Magnética/métodos , Lesões do Ombro/diagnóstico por imagem , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Estudos Retrospectivos , Sensibilidade e Especificidade , Articulação do Ombro/diagnóstico por imagem
10.
Surg Endosc ; 36(2): 1123-1130, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-33629182

RESUMO

BACKGROUND: Endoscopic hemostatic clipping is a safe and efficient treatment used to manage bleeding or perforation during endoscopic submucosal dissection (ESD) for gastric neoplasm. However, the natural history of applied hemoclips during ESD has not been elucidated. As prolonged clip retention limits the use of magnetic resonance imaging and may impede the ulcer healing process, we investigated the factors associated with prolonged hemoclip retention during gastric ESD. METHODS: We retrospectively reviewed 199 patients who underwent gastric ESD with hemoclip application from January 2006 to January 2019. The primary outcome was the prolonged hemoclip retention rate 3 months after ESD. We examined the records of subjects followed at 3, 6, and 12 months and then annually after ESD to monitor clip retention. RESULTS: The prolonged hemoclip retention rate at 3 months was 27.1% (54/199). The risk of hemoclip retention was significantly lower at the antrum (19.6%, P = 0.03). Hemoclips at the angle tended to remain longer than other locations in the stomach (40.6%, P = 0.081) while there was no difference in the number of applied clips depending upon the location of the lesion. By Kaplan-Meier survival analysis, clips at the antrum detached significantly earlier than those at other locations (P = 0.011). CONCLUSIONS: Most of the hemostatic clips attached during ESD were spontaneously removed by 3 months after gastric ESD. However, clips positioned at angle are suspected to have a high probability of prolonged retention. With this in mind, more attention is needed when using hemoclips on angle.


Assuntos
Ressecção Endoscópica de Mucosa , Hemostáticos , Neoplasias Gástricas , Ressecção Endoscópica de Mucosa/efeitos adversos , Ressecção Endoscópica de Mucosa/métodos , Mucosa Gástrica/patologia , Humanos , Estudos Retrospectivos , Fatores de Risco , Neoplasias Gástricas/patologia , Neoplasias Gástricas/cirurgia , Instrumentos Cirúrgicos , Resultado do Tratamento
11.
J Ultrasound Med ; 41(10): 2557-2566, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35040498

RESUMO

OBJECTIVES: To identify sonographic findings that predict clinical outcomes in neonates with portal vein gas. METHODS: This retrospective study included neonates with portal vein gas detected using ultrasound (US). The US images were reviewed by a pediatric radiologist. US findings included the extent and amount of portal vein gas, mesenteric vein gas, intramural gas, bowel wall thickening or thinning, focal discontinuity of the bowel wall, ascites, free intra-abdominal gas, pseudocyst, gas in the other solid organs, and bowel distension. The imaging findings and demographic factors between survivors and non-survivors were statistically compared. RESULTS: The mortality rate was 26% (39 survivors, 14 non-survivors) when iatrogenic and idiopathic causes were excluded. The causes of portal vein gas were determined to be necrotizing enterocolitis (n = 33), bowel distension or obstruction (n = 12), fetal hydrops (n = 4), pneumothorax (n = 3), immediate postoperative state for bowel perforation (n = 1), and umbilical vein catheterization (n = 9) based on surgical findings (n = 20) and clinical information (n = 46). Gas within the mesenteric vein and the other solid organs, gestational age, and birth weight were significantly different between survivors and those who succumbed (P < .05). CONCLUSION: Gas within the mesenteric vein and other solid organs is a US finding that may predict poor outcomes in neonates with portal vein gas.


Assuntos
Enterocolite Necrosante , Doenças do Recém-Nascido , Criança , Enterocolite Necrosante/diagnóstico por imagem , Humanos , Recém-Nascido , Veia Porta/diagnóstico por imagem , Estudos Retrospectivos , Resultado do Tratamento
12.
BMC Palliat Care ; 21(1): 118, 2022 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-35787278

RESUMO

BACKGROUND: A socioculturally appropriate appreciation of dignity is pivotal to the effective provision of care for dying patients. Yet concepts of dignity remain poorly defined. To address this gap in understanding and enhance dignity conserving end-of-life care, a review of current concepts of dignity is proposed. METHODS: To address its primary research question "How do patients conceive the concept of dignity at the end of life?", this review appraises regnant concepts and influences of dignity, and evaluates current dignity conserving practices. To enhance accountability, transparency and reproducibility, this review employs the Ring Theory of Personhood (RToP) as its theoretical lens to guide a Systematic Evidence Based Approach guided Systematic Scoping Review (SSR in SEBA) of patient perspectives of dignity. Three independent teams of reviewers independently analysed included articles from a structured search of PubMed, Embase, PsycINFO, Scopus, CINAHL and Cochrane Databases using thematic and content analyses. The themes and categories identified were compared and combined using the Funnelling Process to create domains that guide the discussion that follows. RESULTS: Seventy-eight thousand five hundred seventy-five abstracts were identified, 645 articles were reviewed, and 127 articles were included. The three domains identified were definitions of dignity, influences upon perceptions of dignity, and dignity conserving care. CONCLUSIONS: This SSR in SEBA affirms the notion that dignity is intimately entwined with self-concepts of personhood and that effective dignity conserving measures at the end of life must be guided by the patient's concept of dignity. This SSR in SEBA posits that such personalised culturally sensitive, and timely support of patients, their family and loved ones may be possible through the early and longitudinal application of a RToP based tool.


Assuntos
Respeito , Assistência Terminal , Morte , Humanos , Pessoalidade , Reprodutibilidade dos Testes
13.
J Clin Ultrasound ; 50(6): 810-813, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35080777

RESUMO

Haddad syndrome is a rare congenital disorder characterized by congenital central hypoventilation syndrome and Hirschsprung disease. Total colonic aganglionosis is a rare and long-segment form of Hirschsprung disease, which is primarily diagnosed using contrast enemas. However, the diagnostic performance of contrast enemas is relatively low, making the diagnosis of total colonic aganglionosis challenging. In neonates, ultrasound may be used as an additional imaging modality for the diagnosis of Hirschsprung disease. We describe the unique sonographic findings of total colonic aganglionosis in a term neonate with failure to pass meconium and respiratory distress, who was subsequently diagnosed with Haddad syndrome.


Assuntos
Doença de Hirschsprung , Apneia do Sono Tipo Central , Doença de Hirschsprung/complicações , Doença de Hirschsprung/diagnóstico por imagem , Humanos , Hipoventilação/congênito , Hipoventilação/diagnóstico , Recém-Nascido , Mecônio , Apneia do Sono Tipo Central/diagnóstico
14.
J Virol ; 94(5)2020 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-31801866

RESUMO

Hepatitis C virus (HCV) is a major etiologic agent of chronic liver diseases. HCV is highly dependent on cellular machinery for viral propagation. Using protein microarray analysis, we previously identified 90 cellular proteins as nonstructural 5A (NS5A) interacting partners. Of these, protein kinase C and casein kinase substrate in neurons protein 2 (PACSIN2) was selected for further study. PACSIN2 belongs to the PACSIN family, which is involved in the formation of caveolae. Protein interaction between NS5A and PACSIN2 was confirmed by pulldown assay and further verified by both coimmunoprecipitation and immunofluorescence assays. We showed that PACSIN2 interacted with domain I of NS5A and the Fer-CIP4 homology (FCH)-Bin/amphiphysin/Rvs (F-BAR) region of PACSIN2. Interestingly, NS5A specifically attenuated protein kinase C alpha (PKCα)-mediated phosphorylation of PACSIN2 at serine 313 by interrupting PACSIN2 and PKCα interaction. In fact, mutation of the serine 313 to alanine (S313A) of PACSIN2 increased protein interaction with NS5A. Silencing of PACSIN2 decreased both viral RNA and protein expression levels of HCV. Ectopic expression of the small interfering RNA (siRNA)-resistant PACSIN2 recovered the viral infectivity, suggesting that PACSIN2 was specifically required for HCV propagation. PACSIN2 was involved in viral assembly without affecting other steps of the HCV life cycle. Indeed, overexpression of PACSIN2 promoted NS5A and core protein (core) interaction. We further showed that inhibition of PKCα increased NS5A and core interaction, suggesting that phosphorylation of PACSIN2 might influence HCV assembly. Moreover, PACSIN2 was required for lipid droplet formation via modulating extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation. Taken together, these data indicate that HCV modulates PACSIN2 via NS5A to promote virion assembly.IMPORTANCE PACSIN2 is a lipid-binding protein that triggers the tubulation of the phosphatidic acid-containing membranes. The functional involvement of PACSIN2 in the virus life cycle has not yet been demonstrated. We showed that phosphorylation of PACSIN2 displayed a negative effect on NS5A and core interaction. The most significant finding is that NS5A prevents PKCα from binding to PACSIN2. Therefore, the phosphorylation level of PACSIN2 is decreased in HCV-infected cells. We showed that HCV NS5A interrupted PKCα-mediated PACSIN2 phosphorylation at serine 313, thereby promoting NS5A-PACSIN2 interaction. We further demonstrated that PACSIN2 modulated lipid droplet formation through ERK1/2 phosphorylation. These data provide evidence that PACSIN2 is a proviral cellular factor required for viral propagation.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Hepacivirus/fisiologia , Domínios e Motivos de Interação entre Proteínas , Proteínas não Estruturais Virais/metabolismo , Montagem de Vírus/fisiologia , Proteínas Adaptadoras de Transdução de Sinal/genética , Linhagem Celular , Regulação Viral da Expressão Gênica , Hepatite C/virologia , Humanos , Imunoprecipitação , Fosforilação , Processamento de Proteína Pós-Traducional , RNA Interferente Pequeno , RNA Viral/metabolismo , Replicação Viral/fisiologia
15.
J Virol ; 94(19)2020 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-32727880

RESUMO

Hepatitis C virus (HCV) exploits cellular proteins to facilitate viral propagation. To identify the cellular factors involved in the HCV life cycle, we previously performed protein microarray assays using either HCV nonstructural 5A (NS5A) protein or core protein as a probe. Interestingly, cellular cortactin strongly interacted with both NS5A and core. Cortactin is an actin-binding protein critically involved in tumor progression by regulating the migration and invasion of cancerous cells. Protein interaction between cortactin and NS5A or core was confirmed by coimmunoprecipitation and immunofluorescence assays. We showed that cortactin interacted with NS5A and core via the N-terminal acidic domain of cortactin. Cortactin expression levels were not altered by HCV infection. Small interfering RNA (siRNA)-mediated knockdown of cortactin dramatically decreased HCV protein expression and infectivity levels, whereas overexpression of cortactin increased viral propagation. Ectopic expression of the siRNA-resistant cortactin recovered the viral infectivity, suggesting that cortactin was specifically required for HCV propagation. We further showed that cortactin was involved in the assembly step without affecting viral entry, HCV internal ribosome entry site (IRES)-mediated translation, and the replication steps of the HCV life cycle. Of note, silencing of cortactin markedly reduced both NS5A and core protein levels on the lipid droplets (LDs), and this effect was reversed by the overexpression of cortactin. Importantly, NS5A and core promoted cell migration by activating the phosphorylation of cortactin at tyrosine residues 421 and 466. Taken together, these data suggest that cortactin is not only involved in HCV assembly but also plays an important role in the cell migration.IMPORTANCE Cortactin is a cytoskeletal protein that regulates cell migration in response to a number of extracellular stimuli. The functional involvement of cortactin in the virus life cycle is not yet fully understood. The most significant finding is that cortactin strongly interacted with both hepatitis C virus (HCV) core and NS5A. Cortactin is involved in HCV assembly by tethering core and NS5A on the lipid droplets (LDs) with no effect on LD biogenesis. It was noteworthy that HCV NS5A and core activated cortactin by phosphorylation at tyrosines 421 and 466 to regulate cell migration. Collectively, our study shows that cortactin is a novel host factor involved in viral production and HCV-associated pathogenesis.


Assuntos
Cortactina/metabolismo , Hepacivirus/fisiologia , Proteínas não Estruturais Virais/metabolismo , Vírion/fisiologia , Montagem de Vírus/fisiologia , Linhagem Celular , Proteínas do Citoesqueleto/metabolismo , Células HEK293 , Hepatite C/virologia , Antígenos da Hepatite C/metabolismo , Humanos , Imunoprecipitação , Fosforilação , RNA Interferente Pequeno/genética , Internalização do Vírus , Replicação Viral
16.
FASEB J ; 34(6): 8686-8701, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32356337

RESUMO

Thyroid hormones are essential for the regulation of energy homeostasis and metabolic processes. However, the relationship between thyroid function and host gut microbial communities is not properly understood. To determine whether and how gut microbiota is associated with thyroid function, metagenomics analysis of the bacterial population in fecal samples of rat models of hyperthyroidism (induced by levothyroxine) and hypothyroidism (induced by propylthiouracil or thyroidectomy) was conducted through 16S rRNA gene sequencing. Our results revealed that all thyroid dysfunction models were definitely established and gut microbial composition varied according to different thyroid functional status. The relative abundance of Ruminococcus was significantly higher in the hyperthyroidism group (HE) vs both the normal and hypothyroidism groups (HO) while S24-7 was significantly higher in the HO group. The population of Prevotellaceae and Prevotella were significantly lower in the HO group vs the normal. Firmicutes and Oscillospira were significantly higher in the SHO (surgery-induced hypothyroidism) group, while Prevotellaceae and Prevotella showed lower abundance in the SHO group than the SHAM group. Present results suggest that thyroid functions may have the potential to influence the profile of gut microbiota and could be used as foundation to investigate interaction mechanism between thyroid and gut microbiome.


Assuntos
Microbioma Gastrointestinal/genética , Glândula Tireoide/microbiologia , Glândula Tireoide/patologia , Animais , Bactérias/genética , Bacteroidetes/genética , Modelos Animais de Doenças , Fezes/microbiologia , Hipotireoidismo/microbiologia , Hipotireoidismo/patologia , Masculino , Metagenômica/métodos , Microbiota/genética , RNA Ribossômico 16S/genética , Ratos , Ratos Sprague-Dawley
17.
Eur Radiol ; 31(9): 6726-6735, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33634322

RESUMO

OBJECTIVES: To evaluate the ability of shear wave elastography (SWE) in diagnosing medial epicondylitis and to compare the diagnostic performance of SWE with that of grey-scale ultrasound (GSU) and strain elastography (SE). METHODS: GSU, SE, and SWE were performed on 61 elbows of 54 patients from March 2018 to April 2019. An experienced radiologist evaluated the GSU findings (swelling, cortical irregularity, hypoechogenicity, calcification, and tear), colour Doppler findings (hyperaemia), SE findings (strain ratio [SR]), and SWE findings (stiffness and shear wave velocity [SWV]). Participants were divided in two groups: patients with clinically diagnosed medial epicondylitis and patients without medial elbow pain. Findings from the two groups were compared, and the receiver operating characteristic (ROC) curves were calculated for significant features. RESULTS: Of the 54 patients, 25 patients with 28 imaged elbows were clinically diagnosed with medial epicondylitis and 29 patients with 33 imaged elbows had no medial elbow pain. Cortical irregularity, hypoechogenicity, calcification, hyperaemia, SR, stiffness, and SWV were significantly different between the two groups. The areas under the ROC curves were 0.838 for hypoechogenicity, 0.948 for SR, 0.999 for stiffness, and 0.999 for SWV. The diagnostic performances of SR, stiffness, and SWV were significantly superior compared to that of hypoechogenicity. However, there were no significant differences among SR, stiffness, and SWV. CONCLUSIONS: SWE can obtain both stiffness and SWV, which are valuable diagnostic tools in the diagnosis of medial epicondylitis. The diagnostic performance of SWE and SE is similar in detecting medial epicondylitis. KEY POINTS: • Shear wave elastography providing stiffness and shear wave velocity showed excellent performance in the diagnosis of medial epicondylitis. • There was no significant difference in the ability of SE and SWE for diagnosing medial epicondylitis.


Assuntos
Técnicas de Imagem por Elasticidade , Articulação do Cotovelo , Tendinopatia do Cotovelo , Articulação do Cotovelo/diagnóstico por imagem , Humanos , Curva ROC , Ultrassonografia
18.
Eur Radiol ; 31(3): 1597-1607, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33128599

RESUMO

OBJECTIVES: To clarify CT findings that predict outcome of conservative treatment in patients with non-strangulated adhesive small bowel obstruction (SBO). METHODS: Unenhanced and contrast-enhanced abdominopelvic CT studies in 189 patients with adhesive SBO who had initial conservative treatment were reviewed. The CT findings included transition zone, beak signs, maximum bowel diameter, bowel diameter ratio, decreased bowel wall enhancement, increased unenhanced bowel wall attenuation, anterior parietal adhesion, bowel wall thickening, closed-loop obstruction, small bowel feces sign, whirl sign, mesenteric haziness, mesenteric, peritoneal fluid, and submucosal edema. These findings were statistically compared according to the success or failure of treatment. RESULTS: Conservative treatment succeeded in 144 patients (76.2%) and failed in 45 patients (23.8%). At multivariate analysis, the lack of small bowel feces sign, focal, diffuse mesenteric haziness, and moderate amount of mesenteric fluid were independent findings predicting failure of conservative treatment, with odds ratios of 5.23, 5.5, 13.55, and 4.89, respectively. The presence of all significant findings showed a high specificity of 97.2% with positive likelihood ratio of 8.8. If CT scans showed none of the three significant findings, the negative predictive value was 97.6% and negative likelihood ratio was 0.08. CONCLUSIONS: The lack of small bowel feces sign, focal, diffuse mesenteric haziness, and moderate amount of mesenteric fluid are independent CT findings predicting the failure of conservative treatment in patients with non-strangulated adhesive SBO. The combination of all CT findings suggests the need for surgery; absence of two or all CT findings should suggest an attempt for conservative treatment. KEY POINTS: • To minimize delayed operation, it is important to identify non-strangulated adhesive small bowel obstruction patients in whom initial conservative treatment is likely to fail. • The lack of small bowel feces sign, the presence of mesenteric haziness, and a moderate amount of mesenteric fluid are independent factors predicting the failure of conservative treatment in patients with non-strangulated adhesive small bowel obstruction. • The combination of all three CT findings suggests the need for surgery; absence of two or all three CT findings should suggest an attempt for conservative treatment.


Assuntos
Tratamento Conservador , Obstrução Intestinal , Adesivos , Humanos , Obstrução Intestinal/diagnóstico por imagem , Estudos Retrospectivos , Tomografia Computadorizada por Raios X
19.
BMC Gastroenterol ; 21(1): 440, 2021 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-34814853

RESUMO

BACKGROUND: The effect of menopausal hormone therapy (MHT) on gastrointestinal (GI) cancers is controversial, and no research has been conducted in the East. This study investigates the association between MHT and GI cancer risks in South Korea. METHODS: A prescription-based cohort study was conducted using the NHIS Sample Cohort (2002-2013) of Korea. We used 1:5 propensity score matching, and 22,577 MHT users and 111,113 non-users were selected. Kaplan-Meier survival curves with log-rank tests were used. Cox proportional hazard models were used to estimate hazard ratios (HR) with 95% confidence intervals (CI). Landmark analysis was used to determine dose-response relationship. RESULTS: The median follow-up was 79.6 of months. Kaplan-Meier survival curve showed less frequent GI cancer diagnoses in MHT users compared to non-users (0.13 vs. 0.16 per 100,000 person-years). Menopausal hormone therapy was associated with decreased incidence of GI cancer (HR = 0.809, 95%CI = 0.691-0.946) and colorectal cancer (CRC) (HR = 0.757, 95%CI = 0.577-0.995). Gastric cancer (GC) incidence showed marginal significance (HR = 0.787, 95%CI = 0.605-1.023). The mortality from GI cancer was lower in MHT users than in non-users (HR = 0.737, 95%CI = 0.547-0.993). The relationship between MHT and GI cancer was stronger with increasing MHT dose in terms of both incidence (Ptrend = 0.0002) and mortality (Ptrend = 0.0064). CONCLUSIONS: The association between MHT use and reduced risks of GI cancers was attributed to CRC and GC and showed a dose-response relationship in a population-based cohort study.


Assuntos
Terapia de Reposição Hormonal , Neoplasias Gástricas , Estudos de Coortes , Humanos , Menopausa , República da Coreia/epidemiologia
20.
Microbiol Immunol ; 65(12): 566-574, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34516008

RESUMO

The performance of the ASTA MicroIDSys system (ASTA), a new matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) system, was evaluated for the identification of viridans group streptococci (VGS) and compared with the results obtained with the Bruker Biotyper system (Bruker Daltonics). A total of 106 Streptococcus reference strains belonging to 24 species from the bacterial strain bank was analyzed using the two MALDI-TOF MS systems. Of the 106 reference strains tested, ASTA MicroIDSys and Bruker Biotyper correctly identified 84.9% and 81.1% at the species level, 100% and 97.2% at the group level and 100% and 98.1% at the genus level, respectively. The difference between the two systems was not statistically significant (P = 0.289). Out of 24 species, 13 species were accurately identified to the species level with 100% accurate identification rates with both systems. The accurate identification rates at the species level of ASTA MicroIDSys and Bruker Biotyper were 100% and 87.5% for the S. anginosus group; 78.4% and 73.5% for the S. mitis group; 91.7% and 91.7% for the S. mutans group; and 100% and 100% for the S. salivarius group, respectively. The ASTA MicroIDSys showed an identification performance equivalent to that of the Bruker Biotyper for VGS. Therefore, it would be useful for the identification of VGS strains in clinical microbiology laboratories.


Assuntos
Bactérias , Estreptococos Viridans , Lasers , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
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