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1.
Proc Natl Acad Sci U S A ; 120(5): e2207091120, 2023 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-36689650

RESUMO

Galectin-4, a member of the galectin family of animal glycan-binding proteins (GBPs), is specifically expressed in gastrointestinal epithelial cells and is known to be able to bind microbes. However, its function in host-gut microbe interactions remains unknown. Here, we show that intracellular galectin-4 in intestinal epithelial cells (IECs) coats cytosolic Salmonella enterica serovar Worthington and induces the formation of bacterial chains and aggregates. Galectin-4 enchains bacteria during their growth by binding to the O-antigen of lipopolysaccharides. Furthermore, the binding of galectin-4 to bacterial surfaces restricts intracellular bacterial motility. Galectin-4 enhances caspase-1 activation and mature IL-18 production in infected IECs especially when autophagy is inhibited. Finally, orally administered S. enterica serovar Worthington, which is recognized by human galectin-4 but not mouse galectin-4, translocated from the intestines to mesenteric lymph nodes less effectively in human galectin-4-transgenic mice than in littermate controls. Our results suggest that galectin-4 plays an important role in host-gut microbe interactions and prevents the dissemination of pathogens. The results of the study revealed a novel mechanism of host-microbe interactions that involves the direct binding of cytosolic lectins to glycans on intracellular microbes.


Assuntos
Galectina 4 , Inflamassomos , Animais , Camundongos , Humanos , Inflamassomos/metabolismo , Galectina 4/metabolismo , Células Epiteliais/metabolismo , Bactérias , Antígenos O/metabolismo
2.
Molecules ; 29(2)2024 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-38257315

RESUMO

Collagen is an important material for biomedical research, but using mammalian tissue-derived collagen carries the risk of zoonotic disease transmission. Marine organisms, such as farmed tilapia, have emerged as a safe alternative source of collagen for biomedical research. However, the tilapia collagen products for biomedical research are rare, and their biological functions remain largely unexamined. In this study, we characterized a commercial tilapia skin collagen using SDS-PAGE and fibril formation assays and evaluated its effects on skin fibroblast adhesion, proliferation, and migration, comparing it with commercial collagen from rat tails, porcine skin, and bovine skin. The results showed that tilapia skin collagen is a type I collagen, similar to rat tail collagen, and has a faster fibril formation rate and better-promoting effects on cell migration than porcine and bovine skin collagen. We also confirmed its application in a 3D culture for kidney cells' spherical cyst formation, fibroblast-induced gel contraction, and tumor spheroid interfacial invasion. Furthermore, we demonstrated that the freeze-dried tilapia skin collagen scaffold improved wound closure in a mouse excisional wound model, similar to commercial porcine or bovine collagen wound dressings. In conclusion, tilapia skin collagen is an ideal biomaterial for biomedical research.


Assuntos
Pesquisa Biomédica , Tilápia , Camundongos , Ratos , Suínos , Animais , Bovinos , Mamíferos , Colágeno/farmacologia , Pele , Modelos Animais de Doenças
3.
Foot Ankle Surg ; 30(3): 191-218, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38278653

RESUMO

BACKGROUND: Treatment of posterior malleolar fracture with plate or screw fixation is still controversial. Plate fixation is considered to have better stability but more soft tissue damage; screw fixation is less invasive and may yields lesser blood loss and surgery time. We conducted this meta-analysis to explore intraoperative and postoperative efficacy between plate and screw fixation in posterior malleolar fractured patients. METHODS: PubMed, Cochrane, Embase, Scopus and Chinese National Knowledge Infrastructure databases were searched in accordance with the Preferred Reporting Items for Systematic Review and Meta-analysis (PRISMA) guidelines. Random-effects model and 95% confidence intervals was used. The outcomes of interest were surgery time, blood loss, length of hospital stay, American Orthopedic Foot and Ankle Score (AOFAS), bone healing time, full weight bearing time, off bed ambulation time, Visual Analogue Scale (VAS), complication rate, and rate of use of syndesmosis screw etc. RESULTS: One randomized clinical trial and fifty-two retrospective cohort studies with a total of 3757 patients (1956 in screw group and 1801 in plate group) were included in the systematic review. Compared to screw group, plate group yielded significantly longer surgery time, more intraoperative blood loss, but shorter length of hospital stay, better AOFAS, better Baird Jackson score, better AOFAS and Baird Jackson excellent-good rate, shorter bone healing time, shorter time enabling full weight bearing, shorter time enabling off bed ambulation, lesser postoperative pain, lesser complication rate, lesser loosening rate, lesser malunion rate, and lesser postoperative osteoarthritis. CONCLUSIONS: Plate fixation is a favorable alternative to screw fixation in posterior malleolar fractured patients. Although plate fixation was at risk of longer surgery time and more blood loss, it provided better postoperative functional outcome, shorter healing, weight bearing and off bed ambulation time and lesser pain compared to screw fixation.


Assuntos
Fraturas do Tornozelo , Placas Ósseas , Parafusos Ósseos , Fixação Interna de Fraturas , Humanos , Fixação Interna de Fraturas/instrumentação , Fraturas do Tornozelo/cirurgia , Duração da Cirurgia
4.
Angew Chem Int Ed Engl ; 63(43): e202407293, 2024 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-39072873

RESUMO

Aminoarenes are important molecules for broad applications in nearly all modern industries that involve chemicals. Direct and site-selective C-H bond amination of arenes provides the most efficient and convenient method to prepare aminoarenes. A main challenge is to selectively install the amino group (or other functional groups) to the distal para-carbon of arenes (especially multi-substituted arenes) during the C-H bond functionalization events. Herein, we address this problem by designing a new strategy via a sequential radical dearomatization/radical amination/rearomatization process for para-selective amination of benzyl alcohols. The para-selectivity of our reaction is completely independent of the electronic and steric properties of the other substituents of the arene substrates. Aminoarenes with many substituents (up to full substitution) and diverse substitution patterns, including those difficult to synthesize previously, could be readily prepared using our protocols. Further exploration of the current strategy shall lead to other challenging C-H functionalization of arenes.

5.
Am J Physiol Lung Cell Mol Physiol ; 324(4): L536-L549, 2023 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-36852927

RESUMO

Interstitial macrophages (IMs) reside in the lung tissue surrounding key structures including airways, vessels, and alveoli. Recent work has described IM heterogeneity during homeostasis, however, there are limited data on IMs during inflammation. We sought to characterize IM origin, subsets, and transcriptomic profiles during homeostasis and lipopolysaccharide (LPS) induced acute lung inflammation. During homeostasis, we used three complementary methods, spectral flow cytometry, single-cell RNA-sequencing, and gene regulatory network enrichment, to demonstrate that IMs can be divided into two core subsets distinguished by surface and transcriptional expression of folate receptor ß (Folr2/FRß). These subsets inhabited distinct niches within the lung interstitium. Within FRß+ IMs we identified a subpopulation marked by coexpression of LYVE1. During acute LPS-induced inflammation, lung IM numbers expand. Lineage tracing revealed IM expansion was due to recruitment of monocyte-derived IMs. At the peak of inflammation, recruited IMs were comprised two unique subsets defined by expression of genes associated with interferon signaling and glycolytic pathways. As recruited IMs matured, they adopted the overall transcriptional state of FRß- resident IMs but retained expression in several origin-specific genes, such as IL-1ß. FRß+ IMs were of near-pure resident origin. Taken together our data show that during LPS-induced inflammation, there are distinct populations of IMs that likely have unique functions. FRΒ+ IMs comprise a stable, resident population, whereas FRß- ΙΜs represent a mixed population of resident and recruited IMs.


Assuntos
Receptor 2 de Folato , Pneumonia , Humanos , Monócitos/metabolismo , Lipopolissacarídeos/farmacologia , Lipopolissacarídeos/metabolismo , Macrófagos/metabolismo , Pneumonia/induzido quimicamente , Pneumonia/genética , Pneumonia/metabolismo , Inflamação/genética , Inflamação/metabolismo , Análise de Sequência de RNA/métodos , Receptor 2 de Folato/metabolismo
6.
PLoS Pathog ; 17(6): e1009602, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-34106992

RESUMO

The CD4+ T cell response is critical to host protection against helminth infection. How this response varies across different hosts and tissues remains an important gap in our understanding. Using IL-4-reporter mice to identify responding CD4+ T cells to Nippostrongylus brasiliensis infection, T cell receptor sequencing paired with novel clustering algorithms revealed a broadly reactive and clonally diverse CD4+ T cell response. While the most prevalent clones and clonotypes exhibited some tissue selectivity, most were observed to reside in both the lung and lung-draining lymph nodes. Antigen-reactivity of the broader repertoires was predicted to be shared across both tissues and individual mice. Transcriptome, trajectory, and chromatin accessibility analysis of lung and lymph-node repertoires revealed three unique but related populations of responding IL-4+ CD4+ T cells consistent with T follicular helper, T helper 2, and a transitional population sharing similarity with both populations. The shared antigen reactivity of lymph node and lung repertoires combined with the adoption of tissue-specific gene programs allows for the pairing of cellular and humoral responses critical to the orchestration of anti-helminth immunity.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Infecções por Strongylida/imunologia , Animais , Pulmão/imunologia , Linfonodos/imunologia , Camundongos , Nippostrongylus , Receptores de Antígenos de Linfócitos T alfa-beta/imunologia , Análise de Célula Única
7.
Proc Natl Acad Sci U S A ; 117(33): 19888-19895, 2020 08 18.
Artigo em Inglês | MEDLINE | ID: mdl-32747552

RESUMO

More than 30% of genes in higher eukaryotes are regulated by RNA polymerase II (Pol II) promoter proximal pausing. Pausing is released by the positive transcription elongation factor complex (P-TEFb). However, the exact mechanism by which this occurs and whether phosphorylation of the carboxyl-terminal domain of Pol II is involved in the process remains unknown. We previously reported that JMJD5 could generate tailless nucleosomes at position +1 from transcription start sites (TSS), thus perhaps enable progression of Pol II. Here we find that knockout of JMJD5 leads to accumulation of nucleosomes at position +1. Absence of JMJD5 also results in loss of or lowered transcription of a large number of genes. Interestingly, we found that phosphorylation, by CDK9, of Ser2 within two neighboring heptad repeats in the carboxyl-terminal domain of Pol II, together with phosphorylation of Ser5 within the second repeat, HR-Ser2p (1, 2)-Ser5p (2) for short, allows Pol II to bind JMJD5 via engagement of the N-terminal domain of JMJD5. We suggest that these events bring JMJD5 near the nucleosome at position +1, thus allowing JMJD5 to clip histones on this nucleosome, a phenomenon that may contribute to release of Pol II pausing.


Assuntos
Quinase 9 Dependente de Ciclina/metabolismo , Histona Desmetilases/metabolismo , RNA Polimerase II/metabolismo , Transcrição Gênica , Linhagem Celular Tumoral , Quinase 9 Dependente de Ciclina/genética , Histona Desmetilases/química , Histona Desmetilases/genética , Humanos , Nucleossomos/genética , Nucleossomos/metabolismo , Fosforilação , Fator B de Elongação Transcricional Positiva/genética , Fator B de Elongação Transcricional Positiva/metabolismo , Regiões Promotoras Genéticas , Ligação Proteica , Domínios Proteicos , RNA Polimerase II/genética
8.
Glycobiology ; 32(1): 73-82, 2022 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-34791227

RESUMO

Enhanced sebocyte proliferation is associated with the pathogenesis of human skin diseases related to sebaceous gland hyperfunction and androgens, which are known to induce sebocyte proliferation, are key mediators of this process. Galectin-12, a member of the ß-galactoside-binding lectin family that is preferentially expressed by adipocytes and functions as an intrinsic negative regulator of lipolysis, has been shown to be expressed by human sebocytes. In this study, we identified galectin-12 as an important intracellular regulator of sebocyte proliferation. Galectin-12 knockdown in the human SZ95 sebocyte line suppressed cell proliferation, and its overexpression promoted cell cycle progression. Inhibition of galectin-12 expression reduced the androgen-induced SZ95 sebocyte proliferation and growth of sebaceous glands in mice, respectively. The mRNA expression of the key cell cycle regulators cyclin A1 (CCNA1) and cyclin-dependent kinase 2CDK2 was reduced in galectin-12 knockdown SZ95 sebocytes, suggesting a pathway of galectin-12 regulation of sebocyte proliferation. Further, galectin-12 enhanced peroxisome proliferator-activated receptor gamma (PPARγ) expression and transcriptional activity in SZ95 sebocytes, consistent with our previous studies in adipocytes. Rosiglitazone, a PPARγ ligand, induced CCNA1 levels, suggesting that galectin-12 may upregulate CCNA1 expression via PPARγ. Our findings suggest the possibility of targeting galectin-12 to treat human sebaceous gland hyperfunction and androgen-associated skin diseases.


Assuntos
Ciclina A1 , Glândulas Sebáceas , Animais , Ciclo Celular/genética , Proliferação de Células , Ciclina A1/metabolismo , Quinase 2 Dependente de Ciclina , Galectinas/genética , Galectinas/metabolismo , Camundongos , Glândulas Sebáceas/metabolismo
9.
Anal Chem ; 94(24): 8732-8739, 2022 06 21.
Artigo em Inglês | MEDLINE | ID: mdl-35678832

RESUMO

Herein, a novel tetrahedral DNA walker with four arms was engineered to travel efficiently on the 3D-tracks via catalyzed hairpin assembly autonomously, realizing the sensitive detection and activity assessment as well as intracellular imaging of apurinic/apyrimidinic endonuclease 1 (APE1). In contrast to traditional DNA walkers, the tetrahedral DNA walker with the rigid 3D framework structure and nonplanar multi-sites walking arms endowed with high collision efficiency, showing a fast walking rate and high nuclease resistance. Impressively, the initial rate of the tetrahedral DNA walker with four arms was 4.54 times faster than that of the free bipedal DNA walker and produced a significant fluorescence recovery in about 40 min, achieving a sensitive detection of APE1 with a low detection limit of 5.54× 10-6 U/µL as well as ultrasensitive intracellular APE1 fluorescence activation imaging. This strategy provides a novel DNA walker for accurate identification of low-abundance cancer biomarker and potential medical diagnosis.


Assuntos
DNA Liase (Sítios Apurínicos ou Apirimidínicos) , DNA , DNA/química , Reparo do DNA , DNA Liase (Sítios Apurínicos ou Apirimidínicos)/metabolismo , Endonucleases , Fluorescência
10.
Anal Chem ; 94(27): 9934-9939, 2022 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-35766464

RESUMO

Herein, Ag@pyrenecarboxaldehyde nanocapsules (Ag@Pyc nanocapsules) as emitters were prepared to construct an ultrasensitive electrochemiluminescence (ECL) biosensor for the detection of the human apurinic/apyrimidinic endonuclease1 (APE1) activity. Ag nanoparticles on the surface of Pyc nanocapsules as coreaction accelerators could significantly promote coreactant peroxydisulfate (S2O82-) to generate massive reactive intermediates of sulfate radical anion (SO4•-), which interacted with the Pyc nanocapsules to achieve a strong ECL response. In addition, with the aid of APE1-triggered 3D DNA machine, trace target could be converted into a large number of mimic targets (MTs), which were positively correlated with the activity of APE1. Consequently, the proposed ECL biosensor realized an ultrasensitive detection of APE1 activity with an exceptional linear working range from 5 × 10-10 to 5 × 10-4 U·µL-1 and a lower limit of detection of 1.36 × 10-11 U·µL-1. This strategy provided a new approach to construct an efficient ternary system for the detection of biomolecules and early diagnosis of diseases.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Nanocápsulas , Técnicas Eletroquímicas , Humanos , Limite de Detecção , Medições Luminescentes , Prata
11.
Int J Mol Sci ; 23(22)2022 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-36430445

RESUMO

Multicellular tumor spheroids and tumoroids are considered ideal in vitro models that reflect the features of the tumor microenvironment. Biomimetic components resembling the extracellular matrix form scaffolds to provide structure to 3-dimensional (3D) culture systems, supporting the growth of both spheroids and tumoroids. Although Matrigel has long been used to support 3D culture systems, batch variations, component complexity, and the use of components derived from tumors are complicating factors. To address these issues, we developed the ACD 3D culture system to provide better control and consistency. We evaluated spheroid and tumoroid formation using the ACD 3D culture system, including the assessment of cell viability and cancer marker expression. Under ACD 3D culture conditions, spheroids derived from cancer cell lines exhibited cancer stem cell characteristics, including a sphere-forming size and the expression of stem cell marker genes. The ACD 3D culture system was also able to support patient-derived primary cells and organoid cell cultures, displaying adequate cell growth, appropriate morphology, and resistance to oxaliplatin treatment. These spheroids could also be used for drug screening purposes. In conclusion, the ACD 3D culture system represents an efficient tool for basic cancer research and therapeutic development.


Assuntos
Neoplasias , Esferoides Celulares , Humanos , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Técnicas de Cultura de Células , Linhagem Celular , Células-Tronco/metabolismo , Microambiente Tumoral
12.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 53(6): 1081-1089, 2022 Nov.
Artigo em Zh | MEDLINE | ID: mdl-36443056

RESUMO

Objective: To study with quantitative computed tomography (QCT) the correlation between grip strength and physical composition and waist and hip circumferences in young people with different body mass indexes (BMIs). Methods: A total of 1310 young people who came to West China Hospital, Sichuan University for physical checkups and underwent chest QCT at our hospital from April to July 2021 were included in the study. Their data were collected and their BMIs were calculated. The subjects were divided according to their BMIs into 4 groups, underweight group (BMI<18.5 kg/m 2), normal-weight group (18.5 kg/m 2≤BMI<24 kg/m 2), overweight group (24 kg/m 2≤BMI<28 kg/m 2), and obesity group (BMI≥28 kg/m 2). The raw data were uploaded to QCT Mindways Pro 6.1 software to be processed for measurement of the fat content (area) of the physical components of the L2 vertebral body, including total adipose tissue (TAT), visceral adipose tissue (VAT), subcutaneous adipose tissue (SAT), and abdominal fat ratio, or VAT/SAT. Grip strength was measured with CAMRY EH101 digital grip dynamometer. Statistical analysis of the data was performed, and the correlations between grip strength and various physical components, waist circumference, and hip circumference in subjects of different BMIs were examined. In addition, stratified analysis of normal-weight and overweight subjects of different age groups was conducted. Results: In the normal-weight group, height, body mass, hip circumference and grip strength were positively correlated with grip strength in males aged 21-40 years, SAT was negatively correlated with grip strength in males aged 36-40 years, and VAT/SAT was positively correlated with grip strength in males aged 36-40 years. In normal-weight females aged 21-25 years, SAT was negatively correlated with grip strength, while VAT and VAT/SAT were positively correlated with grip strength. In normal-weight females aged 26-40 years, height, body mass, and hip circumference were positively correlated with grip strength. In normal-weight females aged 36-40 years, VAT/SAT was positively correlated with grip strength. In overweight males aged 21-25 years, hip circumference and body mass were positively correlated with grip strength. In overweight males aged 26-30 years, TAT, waist-to-hip ratio, and waist-to-height ratio were negatively correlated with grip strength. In overweight males aged 31-40 years, height and body mass were positively correlated with grip strength, while waist-to-hip ratio and waist-to-height ratio were negatively correlated with grip strength. In addition, hip circumference was positively correlated with grip strength in overweight males aged 31-35 years. In overweight females aged 21-25 years, waist circumference, hip circumference, and waist-to-height ratio were positively correlated with grip strength. In overweight females aged 26-30 years, height and body mass were positively correlated with grip strength. In overweight females aged 31-35 years, TAT, SAT, waist circumference, waist-to-hip ratio, and waist-to-height ratio were negatively correlated with grip strength. In overweight females aged 36-40 years, SAT and waist-to-height ratio were negatively correlated with grip strength, while VAT, VAT/SAT, height and body mass were positively correlated with grip strength. The height and body mass of males and females in the underweight group were positively correlated with grip strength, and the hip circumference of females in the underweight group was also positively correlated with grip strength. In the obesity group, TAT, VAT, and waist-to-height ratio were negatively correlated with grip strength in males, but no such correlation was observed in females. Conclusion: There is a close association between abdominal fat content and grip strength in young people with different BMIs, indicating that young people should control abdominal fat content and hip fat content in order to maintain the strength of corresponding muscles.


Assuntos
Sobrepeso , Magreza , Feminino , Masculino , Humanos , Adolescente , Força da Mão , Tomografia Computadorizada por Raios X , Obesidade
13.
J Autoimmun ; 119: 102618, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33714796

RESUMO

BACKGROUND: We investigated the correlation between a history of human papillomavirus (HPV) infection and alopecia areata risk. METHODS: The study cohort comprised 30,001 patients with newly diagnosed HPV infection between 2000 and 2012; and with use of computer-generated randomly numbers, patients not had HPV infection were randomly selected as the comparison cohort. HPV infection cohort were matched to comparison individuals at a 1:1 ratio by age, gender and index year. All study individuals were followed up until they developed alopecia areata, withdraw from the insurance program, lost to follow-up, or until the end of 2013. Cox proportional hazards regression analysis was used to analyze the risk of alopecia areata with hazard ratios (HRs) and 95% confidence intervals (CIs) between the HPV and control cohort. RESULTS: The adjusted hazard ratio (aHR) of alopecia areata for HPV patients relative to controls was 2.55 (95% C.I. = 1.88-3.47) after adjusting sex, age and comorbidities. Subgroup analysis indicated that patients with HPV infections had a significantly greater risk of alopecia areata for both genders, all age subgroups, and those with mental disorder diseases. CONCLUSIONS: A history of HPV infection is associated with the development of subsequent alopecia areata in Taiwanese subjects.


Assuntos
Alopecia em Áreas/epidemiologia , Alopecia em Áreas/etiologia , Infecções por Papillomavirus/complicações , Infecções por Papillomavirus/epidemiologia , Adolescente , Adulto , Idoso , Estudos de Casos e Controles , Comorbidade , Bases de Dados Factuais , Suscetibilidade a Doenças , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Infecções por Papillomavirus/virologia , Vigilância da População , Modelos de Riscos Proporcionais , Medição de Risco , Fatores de Risco , Taiwan/epidemiologia , Adulto Jovem
14.
Int J Syst Evol Microbiol ; 71(12)2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34914571

RESUMO

A novel Gram-stain-positive, strictly anaerobic, elliptical, non-motile and non-flagellated bacterium, designed LZLJ-2T, was isolated from the mud in a fermentation cellar used for the production of Chinese Luzhou-flavour Baijiu. Growth occurred at 28-45 °C (optimum, 37 °C), at pH 6.0-7.0 (optimum, pH 6.0) and with concentrations of NaCl up to 2 % (w/v; optimum, 0 %). On the basis of 16S rRNA gene sequence similarity, strain LZLJ-2T belonged to the genus Thermophilibacter and was most closely related to Thermophilibacter mediterraneus Marseille-P3256T (similarity 96.9 %), Olsenella gallinarum ClaCZ62T (similarity 96.6 %) and Thermophilibacter provencensis Marseille-P2912T (similarity 96.4 %). In addition, strain LZLJ-2T had high similarity to the genus Olsenella, including Olsenella profusa DSM 13989T (similarity 94.9 %), Olsenella umbonata DSM 22620T (similarity 94.9 %), Olsenella uli ATCC 49627T (similarity 94.22 %), Tractidigestivibacter scatoligenes DSM 28304T (similarity 93.9 %) and Paratractidigestivibacter faecalis KCTC 15699T (similarity 93.25 %). Comparative genome analysis showed that orthoANI values between strain LZLJ-2T and Thermophilibacter mediterraneus Marseille-P3256T, Olsenella gallinarum ClaCZ62T, Thermophilibacter provencensis Marseille-P2912T, Olsenella profusa DSM 13989T, Olsenella umbonata DSM 22620T, Olsenella uli ATCC 49627T, Tractidigestivibacter scatoligenes DSM 28304T and Paratractidigestivibacter faecalis KCTC 15699T were 78.68, 78.99, 78.29, 73.40, 74.00, 74.30, 75.08 and 77.23 %, and the genome-to-genome distance values were respectively 22.3, 22.5, 22.4, 19.6, 20.5, 19.7, 20.5 and 21.5 %. The genomic DNA G+C content of strain LZLJ-2T was 65.21 mol%. The predominant cellular fatty acids (>10 %) of strain LZLJ-2T were C18 : 1 cis 9 (33.7 %), C14 : 0 (22.0 %) and C18 : 1 cis 9 DMA (13.5 %). d-Glucose, sucrose, mannose, maltose, lactose (weak), salicin, glycerol (weak), cellobiose and trehalose (weak) could be used by strain LZLJ-2T as sole carbon sources. Enzyme activity results showed positive reactions with valine arylamidase, leucine arylamidase, crystine arylamidase, acid phosphatase, alkaline phosphatase, esterase (C4) (weakly positive), naphthol-AS-BI-phosphohydrolase, α-glucosidase and ß-glucosidase. The major end products of glucose fermentation were lactic acid and acetic acid. It produced skatole from indole acetic acid, and produced p-cresol from modified peptone-yeast extract medium with glucose. Based on the 16S rRNA gene trees as well as the genome core gene tree, it is suggested that Olsenella gallinarum are transferred to genus Thermophilibacter as Thermophilibacter gallinarum comb. nov. Based on phenotypic, genotypic and phylogenetic data, strain LZLJ-2T is considered to represent a novel species of the genus Thermophilibacter, for which the name Thermophilibacter immobilis sp. nov. is proposed. The type strain is LZLJ-2T (=KCTC 25162T=JCM 34224T).


Assuntos
Actinobacteria/classificação , Ácidos Graxos , Fermentação , Filogenia , Microbiologia do Solo , Actinobacteria/isolamento & purificação , Bebidas Alcoólicas , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
15.
Biol Proced Online ; 22: 20, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32884451

RESUMO

BACKGROUND: Ligamentum flavum hypertrophy (LFH) is among the most crucial factors in degenerative lumbar spinal stenosis, which can cause back pain, lower extremity pain, cauda equina syndrome and neurogenic claudication. The exact pathogenesis of LFH remains elusive despite extensive research. Most in vitro studies investigating LFH have been carried out using conventional two-dimensional (2D) cell cultures, which do not resemble in vivo conditions, as they lack crucial pathophysiological factors found in three-dimensional (3D) LFH tissue, such as enhanced cell proliferation and cell cluster formation. In this study, we generated ligamentum flavum (LF) clusters using spheroid cultures derived from primary LFH tissue. RESULTS: The cultured LF spheroids exhibited good viability and growth on an ultra-low attachment 96-well plate (ULA 96-plate) platform according to live/dead staining. Our results showed that the 100-cell culture continued to grow in size, while the 1000-cell culture maintained its size, and the 5000-cell culture exhibited a decreasing trend in size as the culture time increased; long-term culture was validated for at least 28 days. The LF spheroids also maintained the extracellular matrix (ECM) phenotype, i.e., fibronectin, elastin, and collagen I and III. The 2D culture and 3D culture were further compared by cell cycle and Western blot analyses. Finally, we utilized hematoxylin and eosin (H&E) staining to demonstrate that the 3D spheroids resembled part of the cell arrangement in LF hypertrophic tissue. CONCLUSIONS: The developed LF spheroid model has great potential, as it provides a stable culture platform in a 3D model that can further improve our understanding of the pathogenesis of LFH and has applications in future studies.

16.
Plant Biotechnol J ; 17(2): 397-409, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-29992702

RESUMO

Morella rubra, red bayberry, is an economically important fruit tree in south China. Here, we assembled the first high-quality genome for both a female and a male individual of red bayberry. The genome size was 313-Mb, and 90% sequences were assembled into eight pseudo chromosome molecules, with 32 493 predicted genes. By whole-genome comparison between the female and male and association analysis with sequences of bulked and individual DNA samples from female and male, a 59-Kb region determining female was identified and located on distal end of pseudochromosome 8, which contains abundant transposable element and seven putative genes, four of them are related to sex floral development. This 59-Kb female-specific region was likely to be derived from duplication and rearrangement of paralogous genes and retained non-recombinant in the female-specific region. Sex-specific molecular markers developed from candidate genes co-segregated with sex in a genetically diverse female and male germplasm. We propose sex determination follow the ZW model of female heterogamety. The genome sequence of red bayberry provides a valuable resource for plant sex chromosome evolution and also provides important insights for molecular biology, genetics and modern breeding in Myricaceae family.


Assuntos
Evolução Molecular , Genoma de Planta/genética , Myrica/genética , Mapeamento Cromossômico , Flores/genética , Flores/crescimento & desenvolvimento , Flores/fisiologia , Frutas/genética , Frutas/crescimento & desenvolvimento , Frutas/fisiologia , Marcadores Genéticos/genética , Anotação de Sequência Molecular , Myrica/crescimento & desenvolvimento , Myrica/fisiologia , Especificidade de Órgãos , Melhoramento Vegetal
17.
J Phys Chem A ; 122(9): 2479-2484, 2018 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-29457903

RESUMO

We investigated the excited-state proton transfer (ESPT) reaction for 3-cyano-7-azaindole (3CAI) in aqueous solution and in ice. 3CAI undergoes water-catalyzed ESPT in the aqueous solution, giving normal (355 nm) and proton transfer tautomer (∼472 nm) emission bands. Detailed temperature-dependent studies showed that the values of activation free energy (Δ G‡) were similar between N-H and N-D isotopes. Therefore, water-catalyzed ESPT involves a stepwise mechanism incorporating solvation equilibrium ( Keq) to form a 1:1 (molar ratio) water:3CAI cyclic hydrogen-bonded complex as an intermediate, followed by perhaps proton tunneling reaction. In sharp contrast, 3CAI in ice undergoes entirely different photophysical properties, in which 3CAI self-organizes to form a double-hydrogen-bonded dimers at the grain boundary of the polycrystalline. Upon excitation, the dimer proceeds with a fast excited-state double proton transfer reaction, giving rise to solely a tautomer emission (∼450 nm). The distinct difference in ESPT properties between water and ice makes azaindoles feasible for the investigation of water-ice interface property.

18.
Molecules ; 23(8)2018 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-30096898

RESUMO

Amino acid contents and their derived volatile compositions in Cabernet Sauvignon grapes and wines after regulated deficit irrigation (RDI) were investigated during the 2015 and 2016 growing seasons in Yinchuan (NingXia, China). High-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) were used for amino acid and volatile compound analyses. Three RDI strategies were tested: 60% (RDI-1), 70% (RDI-2), and 80% (RDI-3) of grapevine estimated evapotranspiration (ETc), and 100% ETc was used as the control group (CK). RDI-treated vines had lower yields and berry weights with higher total soluble solids than the control treatment. RDI-1 increased proline levels in berries and wines. RDI-2 enhanced tyrosine and asparagine levels in wines. RDI-3 enhanced arginine, alanine, valine, leucine, and isoleucine levels in berries and wines. RDI-2 and RDI-3 increased the concentrations of 2-methyl-1-butyl acetate, benzaldehyde, 3-methyl-1-pentanol, and 3-methyl-1-butanol in wines. The accumulation of volatile compounds was closely related to the amino acid concentrations-especially isoleucine, valine, and leucine-in grapes. Our results showed that RDI treatments altered amino acid concentrations and their derived volatile compositions in wines.


Assuntos
Irrigação Agrícola , Aminoácidos/análise , Frutas/química , Vitis/química , Compostos Orgânicos Voláteis/análise , Vinho/análise , Análise por Conglomerados , Tempo (Meteorologia)
19.
Angew Chem Int Ed Engl ; 57(18): 5020-5024, 2018 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-29468788

RESUMO

The compound 6-azaindole undergoes self-assembly by formation of N(1)-H⋅⋅⋅N(6) hydrogen bonds (H bonds), forming a cyclic, triply H-bonded trimer. The formation phenomenon is visualized by scanning tunneling microscopy. Remarkably, the H-bonded trimer undergoes excited-state triple proton transfer (ESTPT), resulting in a proton-transfer tautomer emission maximized at 435 nm (325 nm of the normal emission) in cyclohexane. Computational approaches affirm the thermodynamically favorable H-bonded trimer formation and the associated ESTPT reaction. Thus, nearly half a century after Michael Kasha discovered the double H-bonded dimer of 7-azaindole and its associated excited-state double-proton-transfer reaction, the triply H-bonded trimer formation of 6-azaindole and its ESTPT reaction are demonstrated.

20.
Cytometry A ; 87(1): 49-60, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25352187

RESUMO

A high throughput 3D image cytometer have been developed that improves imaging speed by an order of magnitude over current technologies. This imaging speed improvement was realized by combining several key components. First, a depth-resolved image can be rapidly generated using a structured light reconstruction algorithm that requires only two wide field images, one with uniform illumination and the other with structured illumination. Second, depth scanning is implemented using the high speed remote depth scanning. Finally, the large field of view, high NA objective lens and the high pixelation, high frame rate sCMOS camera enable high resolution, high sensitivity imaging of a large cell population. This system can image at 800 cell/sec in 3D at submicron resolution corresponding to imaging 1 million cells in 20 min. The statistical accuracy of this instrument is verified by quantitatively measuring rare cell populations with ratio ranging from 1:1 to 1:10(5) . © 2014 International Society for Advancement of Cytometry.


Assuntos
Algoritmos , Citometria por Imagem/instrumentação , Imageamento Tridimensional/instrumentação , Microscopia/instrumentação , Animais , Fibroblastos/ultraestrutura , Corantes Fluorescentes , Citometria por Imagem/métodos , Imageamento Tridimensional/métodos , Rim/ultraestrutura , Lentes , Luz , Iluminação , Camundongos , Microscopia/métodos , Cervo Muntjac , Fatores de Tempo
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