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1.
Apoptosis ; 29(1-2): 142-153, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37698736

RESUMO

Increasing data and literature have illustrated that tumor immune escape represents a major source of tumor formation and recrudesce. Besides, novel findings also indicate that RNA N6-methyladenosine (m6A) participates in the human cancer immune escape. Here, our study investigated the functions of m6A reader YTHDF1 in prostate cancer (PCa) immune response and explored the functional mechanism. Results reported that YTHDF1 up-regulated in PCa samples and was closely correlated to poor clinical prognosis. Functionally, YTHDF1 inhibited the killing activity of CD8 + T cells to PCa cells, and moreover mitigated the ferroptosis. Mechanistically, PD-L1 acted as the target of YTHDF1, and YTHDF1 upregulated the transcriptional activity of PD-L1 mRNA. Collectively, YTHDF1 promoted functional PD-L1 partially through enhancing its transcriptional stability, which was necessary for PCa cells to evade effector T cell cytotoxicity and CD8 + T cells mediated ferroptosis. In conclusion, these findings indicate that YTHDF1 represses the CD8 + T cell-mediated antitumor immunity and ferroptosis in PCa via m6A-PD-L1 manner, which may provide novel insight for PCa immunotherapy.


Assuntos
Adenosina/análogos & derivados , Ferroptose , Neoplasias da Próstata , Humanos , Masculino , Ferroptose/genética , Antígeno B7-H1/genética , Apoptose , Neoplasias da Próstata/genética , Linfócitos T CD8-Positivos , Proteínas de Ligação a RNA/genética
2.
Wound Repair Regen ; 32(1): 47-54, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38087425

RESUMO

The aim of this case-control study was to explore the potential risk factors for venous ulceration in patients with varicose veins of lower extremities and to establish a simplified diagnostic score model. Seventy subjects with varicose veins of lower extremities and venous ulceration were compared with 1164 controls with varicose veins of lower extremities and no history of venous ulceration. Stepwise multivariate logistic regression analysis was used to identify the risk factors for venous ulceration. The steps in developing the diagnostic score model were based on the Framingham Heart study. The area under the receiver operating characteristic curve (AUC) was calculated to assess the diagnostic ability of the diagnostic score model. Multivariate analysis showed that men, overweight, obesity, longer duration varicose veins, deep venous valve insufficiency, low lymphocyte counts, and high fibrinogen content were independently associated with an increased risk of venous ulceration. The AUC for the diagnostic score model was 0.75, which indicated good discriminatory ability. Special attention should be paid to the high-risk group of patients with lower extremity varicose veins. The diagnostic score model might be a useful screening tool for clinicians, policy makers, and patients.


Assuntos
Úlcera Varicosa , Varizes , Insuficiência Venosa , Masculino , Humanos , Estudos de Casos e Controles , Cicatrização , Varizes/complicações , Varizes/diagnóstico , Úlcera Varicosa/diagnóstico , Insuficiência Venosa/complicações , Insuficiência Venosa/diagnóstico , Fatores de Risco , Extremidade Inferior
3.
BMC Pediatr ; 24(1): 291, 2024 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-38689221

RESUMO

BACKGROUND: X-linked agammaglobulinemia (XLA) is a primary immunodeficiency disease caused by mutations in the Bruton tyrosine kinase (BTK) gene. Individuals diagnosed with XLA are at an increased risk of developing autoimmune diseases. However, renal involvement are rare in cases of XLA. CASE PRESENTATION: In this report, we discussed a specific case involving a 6-year-old boy with XLA who experienced recurrent upper respiratory tract infections since the age of one. He presented with symptoms of hematuria and proteinuria, and renal pathology confirmed the presence of immunoglobulin (Ig) A nephropathy. Treatment comprised glucocorticoids, mycophenolate mofetil, and intermittent intravenous immunoglobulin replacement therapy. Consequently, there was a remission of proteinuria and a partial improvement in hematuria. CONCLUSIONS: In this study, we describe the first case of IgA nephropathy associated with XLA. This is an interesting phenotype found in XLA, and it provides valuable insights into the process of autoimmunity and the regulation of immune function in individuals with XLA. Based on our findings, we recommend the evaluation of immunoglobulin levels in patients diagnosed with IgA nephropathy.


Assuntos
Agamaglobulinemia , Doenças Genéticas Ligadas ao Cromossomo X , Glomerulonefrite por IGA , Humanos , Agamaglobulinemia/complicações , Agamaglobulinemia/diagnóstico , Agamaglobulinemia/genética , Masculino , Glomerulonefrite por IGA/complicações , Glomerulonefrite por IGA/diagnóstico , Doenças Genéticas Ligadas ao Cromossomo X/complicações , Doenças Genéticas Ligadas ao Cromossomo X/genética , Doenças Genéticas Ligadas ao Cromossomo X/diagnóstico , Criança , Imunoglobulinas Intravenosas/uso terapêutico , Glucocorticoides/uso terapêutico , Ácido Micofenólico/uso terapêutico , Imunossupressores/uso terapêutico
4.
J Neurosci ; 2022 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-35667851

RESUMO

Classical forward and reverse mouse genetics require germline mutations and, thus, are unwieldy to study sleep functions of essential genes or redundant pathways. It is also time-consuming to conduct electroencephalogram/electromyogram-based mouse sleep screening owing to labor-intensive surgeries and genetic crosses. Here, we describe a highly accurate SleepV (video) system and adeno-associated virus (AAV)-based adult brain chimeric (ABC)-expression/knockout (KO) platform for somatic genetics analysis of sleep in adult male or female mice. A pilot ABC screen identifies CREB and CRTC1, of which constitutive or inducible expression significantly reduces quantity and/or quality of non-rapid eye movement sleep. Whereas ABC-KO of exon 13 of Sik3 by AAV-Cre injection in Sik3-E13flox/flox adult mice phenocopies Sleepy (Sik3Slp/+) mice, ABC-CRISPR of Slp/Sik3 reverses hypersomnia of Sleepy mice, indicating a direct role of SLP/SIK3 kinase in sleep regulation. Multiplex ABC-CRISPR of both orexin/hypocretin receptors causes narcolepsy episodes, enabling one-step analysis of redundant genes in adult mice. Therefore, this somatic genetics approach should facilitate high-throughput analysis of sleep regulatory genes, especially for essential or redundant genes, in adult mice by skipping mouse development and minimizing genetic crosses.SIGNIFICANCE STATEMENTThe molecular mechanisms of mammalian sleep regulation remain unclear. Classical germline mouse genetics are unwieldy to study sleep functions of essential genes or redundant pathways. The EEG/EMG-based mouse sleep screening is time-consuming owing to labor-intensive surgeries and lengthy genetic crosses. To overcome these "bottlenecks", we developed a highly accurate video-based sleep analysis system and adeno-associated virus-mediated ABC-expression/knockout platform for somatic genetics analysis of sleep in adult mice. These methodologies facilitate rapid identification of sleep regulatory genes, but also efficient mechanistic studies of the molecular pathways of sleep regulation in mice.

5.
Mol Cell ; 59(5): 807-18, 2015 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-26257286

RESUMO

Assembly of the RNA-induced silencing complex (RISC) requires formation of the RISC loading complex (RLC), which contains the Dicer-2 (Dcr-2)-R2D2 complex and recruits duplex siRNA to Ago2 in Drosophila melanogaster. However, the precise composition and action mechanism of Drosophila RLC remain unclear. Here we identified the missing factor of RLC as TATA-binding protein-associated factor 11 (TAF11) by genetic screen. Although it is an annotated nuclear transcription factor, we found that TAF11 also associated with Dcr-2/R2D2 and localized to cytoplasmic D2 bodies. Consistent with defective RLC assembly in taf11(-/-) ovary extract, we reconstituted the RLC in vitro using the recombinant Dcr-2-R2D2 complex, TAF11, and duplex siRNA. Furthermore, we showed that TAF11 tetramer facilitates Dcr-2-R2D2 tetramerization to enhance siRNA binding and RISC loading activities. Together, our genetic and biochemical studies define the molecular nature of the Drosophila RLC and elucidate a cytoplasmic function of TAF11 in organizing RLC assembly to enhance RNAi efficiency.


Assuntos
Proteínas de Drosophila/química , Proteínas de Drosophila/metabolismo , Interferência de RNA , Complexo de Inativação Induzido por RNA/química , Complexo de Inativação Induzido por RNA/metabolismo , Fatores Associados à Proteína de Ligação a TATA/química , Fatores Associados à Proteína de Ligação a TATA/metabolismo , Animais , Animais Geneticamente Modificados , Proteínas Argonautas/química , Proteínas Argonautas/genética , Proteínas Argonautas/metabolismo , Sistemas CRISPR-Cas , Linhagem Celular , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Drosophila melanogaster/virologia , Feminino , Técnicas de Inativação de Genes , Genes de Insetos , Masculino , Modelos Biológicos , Mutagênese , Estrutura Quaternária de Proteína , RNA Helicases/química , RNA Helicases/genética , RNA Helicases/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Proteínas de Ligação a RNA/química , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Complexo de Inativação Induzido por RNA/genética , Ribonuclease III/química , Ribonuclease III/genética , Ribonuclease III/metabolismo , Fatores Associados à Proteína de Ligação a TATA/genética
6.
J Environ Manage ; 325(Pt A): 116430, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36240640

RESUMO

Although the coupling process of microbial fuel cell (MFC) and activated sludge is widely used for organic matter removal and electric energy recovery, the problem of high effluent nitrate still exists due to the lack of influent carbon source. Herein, a poly (butanediol succinate) (PBS) assembled MFC was established in an aerobic granular sludge (AGS) bioreactor for simultaneous promoting nitrogen removal and electricity generation. Compared to AGS-Control group, the total inorganic nitrogen (TIN) and COD removal efficiencies of AGS-MFC group were improved to 84.3 ± 2.6% and 93.5 ± 0.5% after 100-days operation. The average output voltage and the maximum power density of the MFC module were 223.7 mV and 59.6 mW/m2, respectively. Through high-throughput sequencing analysis, Thauera-related denitrifying bacteria had the highest relative abundances (20.0% and 31.4%) in both bioreactors. The relative abundance of Nitrosomonas-related ammonia oxidizing bacteria (AOB) in AGS-MFC (1.8%) was enriched than AGS-Control (1.1%). In MFC module, Thauera (16.2%) with denitrification and power generation was dominant in anodic biofilms under PBS enhancement. This study provides scientific basis for the application of submersible MFC enhanced deep nitrogen removal under aerobic conditions.


Assuntos
Fontes de Energia Bioelétrica , Esgotos , Esgotos/microbiologia , Fontes de Energia Bioelétrica/microbiologia , Nitrogênio , Desnitrificação , Carbono , Reatores Biológicos/microbiologia
7.
Molecules ; 28(22)2023 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-38005185

RESUMO

Glycosidases are essential for the industrial production of functional oligosaccharides and many biotech applications. A novel ß-galactosidase/α-L-arabinopyranosidase (PpBGal42A) of the glycoside hydrolase family 42 (GH42) from Paenibacillus polymyxa KF-1 was identified and functionally characterized. Using pNPG as a substrate, the recombinant PpBGal42A (77.16 kD) was shown to have an optimal temperature and pH of 30 °C and 6.0. Using pNPαArap as a substrate, the optimal temperature and pH were 40 °C and 7.0. PpBGal42A has good temperature and pH stability. Furthermore, Na+, K+, Li+, and Ca2+ (5 mmol/L) enhanced the enzymatic activity, whereas Mn2+, Cu2+, Zn2+, and Hg2+ significantly reduced the enzymatic activity. PpBGal42A hydrolyzed pNP-ß-D-galactoside and pNP-α-L-arabinopyranoside. PpBGal42A liberated galactose from ß-1,3/4/6-galactobiose and galactan. PpBGal42A hydrolyzed arabinopyranose at C20 of ginsenoside Rb2, but could not cleave arabinofuranose at C20 of ginsenoside Rc. Meanwhile, the molecular docking results revealed that PpBGal42A efficiently recognized and catalyzed lactose. PpBGal42A hydrolyzes lactose to galactose and glucose. PpBGal42A exhibits significant degradative activity towards citrus pectin when combined with pectinase. Our findings suggest that PpBGal42A is a novel bifunctional enzyme that is active as a ß-galactosidase and α-L-arabinopyranosidase. This study expands on the diversity of bifunctional enzymes and provides a potentially effective tool for the food industry.


Assuntos
Paenibacillus polymyxa , Paenibacillus , Paenibacillus polymyxa/metabolismo , Lactose , Simulação de Acoplamento Molecular , Galactose , Glicosídeo Hidrolases/metabolismo , Clonagem Molecular , beta-Galactosidase/metabolismo , Concentração de Íons de Hidrogênio , Especificidade por Substrato , Paenibacillus/genética , Paenibacillus/metabolismo
8.
Molecules ; 28(23)2023 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-38067658

RESUMO

Ferroptosis is a form of regulated cell death that is characterized by the accumulation of iron-dependent lipid peroxides. The regulation of ferroptosis involves both non-enzymatic reactions and enzymatic mechanisms. Natural products have demonstrated potential effects on various enzymes, including GPX4, HO-1, NQO1, NOX4, GCLC, and GCLM, which are mainly involved in glutathione metabolic pathway or oxidative stress regulation, and ACSL3 and ACSL4, which mainly participate in lipid metabolism, thereby influencing the regulation of ferroptosis. In this review, we have provided a comprehensive overview of the existing literature pertaining to the effects of natural products on enzymes involved in ferroptosis and discussed their potential implications for the prevention and treatment of ferroptosis-related diseases. We also highlight the potential challenge that the majority of research has concentrated on investigating the impact of natural products on the expression of enzymes involving ferroptosis while limited attention is given to the regulation of enzyme activity. This observation underscores the considerable potential and scope for exploring the influence of natural products on enzyme activity.


Assuntos
Produtos Biológicos , Ferroptose , Produtos Biológicos/farmacologia , Glutationa , Ferro , Metabolismo dos Lipídeos
9.
J Anim Physiol Anim Nutr (Berl) ; 107(1): 275-285, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36262057

RESUMO

This study evaluated the effects of different dietary metabolizable energy (ME) concentrations on the meat quality, carcass traits, volatile flavour and lipid metabolism-related gene expression levels in yellow-feathered chickens. In total, 600 Huxu female chickens aged 90 days were randomly assigned to six dietary treatments, each with 10 replicates of 10 birds. During the finisher phase, the birds were fed diets containing 2880 (low), 2940, 3000, 3060, 3120 and 3180 (high) kcal ME/kg. The results showed that the average daily gain of chickens increased as the dietary ME concentration increased, while the feed to gain improved (p < 0.05), and the intramuscular fat content of breast muscle increased (p < 0.05). The energy concentration had no effect on the breast muscle pH (45 min and 24 h), colour parameter (L*) or percentage of drip loss (p > 0.05), but the shear force values decreased significantly (p < 0.05). The diameter and area of the breast muscle fiber decreased and the muscle fibre density increased as the dietary ME concentration increased (p < 0.05). The highest ME concentration (3180 kcal) increased the percentages of aldehydes (hexanal, heptanal, 2,4-nonadienal, octanal, nonanal and 2-decenal), alcohols (2-nonen-1-ol, trans-2-undecen-1-ol, 7-hexadecenal, 2-hexyl-1-decanoal and n-nonadecanol-1,3,7,11-trimethyl-1-dodecanol), alkanes (2,6-dimethyl-heptadecane) and carboxylic acids (9-hexadecenoic acid), but reduced the percentages of octadecanal, octadecane, heneicosane and tetradecanal (p < 0.05). In addition, the mRNA gene expression levels of fatty acid-binding protein 3 and apolipoprotein B were significantly upregulated in the liver, whereas that of cholesteryl ester transfer protein was significantly downregulated. In conclusion, increasing the ME diet to 3180 kcal/kg significantly improved the quality and flavour of the meat from yellow-feathered broilers. our finding may help poultry producers to improve the taste of meat by regulating genes related to lipid metabolism, thereby achieving the flavour and taste characteristics preferred by consumers.


Assuntos
Galinhas , Suplementos Nutricionais , Animais , Feminino , Galinhas/fisiologia , Metabolismo dos Lipídeos , Dieta/veterinária , Carne/análise , Expressão Gênica , Ração Animal/análise
10.
Int Wound J ; 21(3): e14492, 2023 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-37989716

RESUMO

Eschar dermabrasion is an easy, cost-effective and dependable technique for debriding deep partial-thickness burn wounds, highly suitable for paediatric scalds. Postoperative dressing plays a crucial role in the subsequent healing process. While allogenic skin (AGS) has long been considered as the optimal coverage for abraded burn wounds by Chinese burn specialists, its clinical application on children has encountered challenges. In recent years, our department has observed promising results in the application of bacterial cellulose dressing on paediatric burn wounds after dermabrasion surgery. This study aimed to retrospectively review qualified cases from the past 5 years and categorize them into two groups: 201 cases in the AGS group and 116 cases in the bacterial cellulose dressing (BCD) group. Upon statistical analysis, no differences were oberved between the groups in terms of demographic information and wound characteristics. However, the BCD group had a significantly longer surgery time (44.3 ± 7.0 min vs. 31.5 ± 6.1 min, p < 0.01) and shorter healing time (19.6 ± 2.2 days vs. 24.4 ± 4.3 days, p < 0.01) compared to the AGS group. Moreover, the BCD group required fewer dressing changes (3.5 ± 0.8 vs. 6.7 ± 2.1, p < 0.01) and demonstrated lower rates of skin grafting (10/116 vs. 46/201, p = 0.036). In conclusion, our findings suggest that the bacterial cellulose material may serve as an optimal coverage option for paediatric abraded scald wounds.

11.
Int Wound J ; 2023 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-37743574

RESUMO

The current methods for the prediction of mortality and amputation for inpatients with diabetic foot (DF) use only conventional, simple variables, which limits their performance. Here, we used a random survival forest (RSF) model and multicomponent variables to improve the prediction of mortality and amputation for these patients. We performed a retrospective cohort study of 175 inpatients with DF who were recruited between 2014 and 2021. Thirty-one predictors in six categories were considered as potential covariates. Seventy percent (n = 122) of the participants were randomly selected to constitute a training set, and 30% (n = 53) were assigned to a testing set. The RSF model was used to screen appropriate variables for their value as predictors of 2-year all-cause mortality and amputation, and a multicomponent prediction model was established. Model performance was evaluated using the area under the curve (AUC) and the Hosmer-Lemeshow test. The AUCs were compared using the Delong test. Seventeen variables were selected to predict mortality and 23 were selected to predict amputation. Uric acid and alanine transaminase were the top two most useful variables for the prediction of mortality, whereas urine protein and platelet were the top variables for the prediction of amputation. The AUCs were 0.913 and 0.851 for the prediction of mortality for the training and testing sets, respectively; and the equivalent AUCs were 0.963 and 0.893 for the prediction of amputation. There were no significant differences between the AUCs for the training and testing sets for both the mortality and amputation models. These models showed a good degree of fit. Thus, the RSF model can predict mortality and amputation in inpatients with DF. This multicomponent prediction model could help clinicians consider predictors of different dimensions to effectively prevent DF from clinical outcomes .

12.
Physiol Mol Biol Plants ; 29(11): 1647-1661, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-38162922

RESUMO

The ascorbate-glutathione (AsA-GSH) cycle is essential for detoxifying reactive oxygen species (ROS) under environmental stresses. The toxicity of aluminum (Al) limits the growth and performance of cultivated plants in acidic soil. However, there is limited information available on the relationship between arbuscular mycorrhizal symbiosis and the AsA-GSH cycle in host plants under Al stress. This study aimed to examine the impact of arbuscular mycorrhizal fungi (AMF), specifically Funneliformis mosseae, on the growth, antioxidant enzymes, components of the AsA-GSH cycle, and stress response gene expressions in white clover (Trifolium repens L.) under Al stress. Our findings demonstrate that AMF inoculation significantly reduced Al accumulation and increased phosphorus (P) content in the roots of white clover, thereby promoting plant biomass accumulation and mycorrhizal colonization under Al stress. AMF effectively scavenged Al-induced ROS (H2O2 and O2-) by enhancing the activities of antioxidant enzymes, including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), as well as the components of the AsA-GSH cycle (e.g., enzymes and antioxidants) in the leaves and roots of white clover plants. Additionally, the mitigating effect of AMF was associated with the upregulation of genes involved in P transport (PHO1-2 and PHT1-7), the AsA-GSH pathway (GST-2 and APX-2), and Al stress (ALMT1) in white clover roots compared to control plants. Principal component analysis revealed that 65.9% of the total variance was explained by the first principal component. Dry mass showed a positive correlation with POD and P content, while exhibiting a highly negative correlation with ROS, antioxidant physiology index, Al content, and the expression of related genes in white clover. Overall, this study suggests that AMF enhances the tolerance of white clover to Al stress by improving P uptake and strengthening the AsA-GSH cycle.

13.
Angew Chem Int Ed Engl ; 62(22): e202300424, 2023 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-36929518

RESUMO

We report a ligand-controlled CoII -catalyzed C(sp3 )-C(sp3 ) coupling hydroalkylation for direct and ß-selective synthesis of 2-deoxy-C-glycosides from glycals. This reaction proceeds by a radical pathway for alkyl halide activation and is ß-selective through ligand control. This approach may inspire the development of further stereoselective coupling reactions with potential application in the field of carbohydrates.

14.
Mol Microbiol ; 116(6): 1433-1448, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34738268

RESUMO

Chlamydia muridarum actively grows in murine mucosae and is a representative model of human chlamydial genital tract disease. In contrast, C. trachomatis infections in mice are limited and rarely cause disease. The factors that contribute to these differences in host adaptation and specificity remain elusive. Overall genomic similarity leads to challenges in the understanding of these significant differences in tropism. A region of major genetic divergence termed the plasticity zone (PZ) has been hypothesized to contribute to the host specificity. To evaluate this hypothesis, lateral gene transfer was used to generate multiple hetero-genomic strains that are predominately C. trachomatis but have replaced regions of the PZ with those from C. muridarum. In vitro analysis of these chimeras revealed C. trachomatis-like growth as well as poor mouse infection capabilities. Growth-independent cytotoxicity phenotypes have been ascribed to three large putative cytotoxins (LCT) encoded in the C. muridarum PZ. However, analysis of PZ chimeras supported that gene products other than the LCTs are responsible for cytopathic and cytotoxic phenotypes. Growth analysis of associated chimeras also led to the discovery of an inclusion protein, CTL0402 (CT147), and homolog TC0424, which was critical for the integrity of the inclusion and preventing apoptosis.


Assuntos
Infecções por Chlamydia/microbiologia , Chlamydia muridarum/genética , Chlamydia trachomatis/genética , Transferência Genética Horizontal , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Chlamydia muridarum/metabolismo , Chlamydia trachomatis/metabolismo , Feminino , Variação Genética , Humanos , Camundongos Endogâmicos C57BL
15.
BMC Biotechnol ; 22(1): 11, 2022 03 20.
Artigo em Inglês | MEDLINE | ID: mdl-35307009

RESUMO

BACKGROUND: Feathers are the most abundant agricultural waste produced by poultry farms. The accumulation of a large number of feathers not only seriously pollutes the environment but also causes the waste of protein resources. The degradation of feather waste by keratinase-producing strains is currently a promising method. Therefore, screening high-producing keratinase strains from marine environment and studying the fermentation conditions, enzymatic properties and feather degradation mechanism are crucial for efficient degradation of feathers. RESULTS: A novel efficient feather-degrading bacteria, Gxun-17, isolated from the soil sample of a marine duck farm of Beibu Gulf in Guangxi, China, was identified as Bacillus tropicus. The optimum fermentation conditions were obtained by single factor and orthogonal tests as follows: feather concentration of 15 g/L, maltose concentration of 10.0 g/L, MgSO4 concentration of 0.1 g/L, initial pH of 7.0 and temperature of 32.5 °C. The strain completely degraded the feathers within 48 h, and the highest keratinase activity was 112.57 U/mL, which was 3.18-fold that obtained with the basic medium (35.37 U/mL). Detecting the keratinase activity and the content of sulphur-containing compounds in the fermentation products showed that the degradation of feathers by the strain might be a synergistic effect of the enzyme and sulphite. The keratinase showed optimal enzyme activity at pH 7.0 and temperature of 60 °C. The keratinase had the best performance on the casein substrate. When casein was used as the substrate, the Km and Vmax values were 15.24 mg/mL and 0.01 mg/(mL·min), respectively. Mg2+, Ca2+, K+, Co2+, Al3+, phenylmethylsulphonyl fluoride and isopropanol inhibited keratinase activity, which indicated that it was a serine keratinase. Conversely, the keratinase activity strongly increased with the addition of Mn2+ and ß-mercaptoethanol. CONCLUSIONS: A novel feather-degrading B. tropicus Gxun-17 was obtained from marine environment. The strain adapted the extreme conditions such as low temperature, high salt and high pressure. Thus, the keratinase had high activity, wide range of temperature and pH, salt tolerance and other characteristics, which had potential application value.


Assuntos
Caseínas , Plumas , Animais , Bacillus , Caseínas/metabolismo , Galinhas/metabolismo , China , Plumas/química , Concentração de Íons de Hidrogênio , Queratinas/análise , Queratinas/química , Queratinas/metabolismo , Peptídeo Hidrolases/metabolismo , Temperatura
16.
Cell Commun Signal ; 20(1): 52, 2022 04 12.
Artigo em Inglês | MEDLINE | ID: mdl-35413847

RESUMO

BACKGROUND: Skin innervation is crucial for normal wound healing. However, the relationship between nerve receptors and wound healing and the intrinsic mechanism remains to be further identified. In this study, we investigated the role of a calcitonin gene-related peptide (CGRP) receptor component, receptor activity-modifying protein 1 (RAMP1), in mouse skin fibroblast (MSF) proliferation. METHODS: In vivo, Western blotting and immunohistochemical (IHC) staining of mouse skin wounds tissue was used to detect changes in RAMP1 expression. In vitro, RAMP1 was overexpressed in MSF cell lines by infection with Tet-On-Flag-RAMP1 lentivirus and doxycycline (DOX) induction. An IncuCyte S3 Live-Cell Analysis System was used to assess and compare the proliferation rate differences between different treatment groups. Total protein and subcellular extraction Western blot analysis, quantitative real-time-polymerase chain reaction (qPCR) analysis, and immunofluorescence (IF) staining analysis were conducted to detect signalling molecule expression and/or distribution. The CUT & RUN assay and dual-luciferase reporter assay were applied to measure protein-DNA interactions. RESULTS: RAMP1 expression levels were altered during skin wound healing in mice. RAMP1 overexpression promoted MSF proliferation. Mechanistically, total Yes-associated protein (YAP) and nuclear YAP protein expression was increased in RAMP1-overexpressing MSFs. RAMP1 overexpression increased inhibitory guanine nucleotide-binding protein (G protein) α subunit 3 (Gαi3) expression and activated downstream protein kinase A (PKA), and both elevated the expression of cyclic adenosine monophosphate (cAMP) response element-binding protein (CREB) and activated it, promoting the transcription of YAP, elevating the total YAP level and promoting MSF proliferation. CONCLUSIONS: Based on these data, we report, for the first time, that changes in the total RAMP1 levels during wound healing and RAMP1 overexpression alone can promote MSF proliferation via the Gαi3-PKA-CREB-YAP axis, a finding critical for understanding RAMP1 function, suggesting that this pathway is an attractive and accurate nerve target for skin wound treatment. Video Abstract.


Assuntos
Proteínas Quinases Dependentes de AMP Cíclico , Subunidades alfa Gi-Go de Proteínas de Ligação ao GTP , Proteína 1 Modificadora da Atividade de Receptores , Transdução de Sinais , Pele , Proteínas de Sinalização YAP , Animais , Proliferação de Células , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Fibroblastos/citologia , Fibroblastos/metabolismo , Subunidades alfa Gi-Go de Proteínas de Ligação ao GTP/metabolismo , Camundongos , Proteína 1 Modificadora da Atividade de Receptores/genética , Proteína 1 Modificadora da Atividade de Receptores/metabolismo , Pele/citologia , Pele/metabolismo , Proteínas de Sinalização YAP/metabolismo
17.
Arch Microbiol ; 204(6): 298, 2022 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-35513460

RESUMO

A novel bacterium of the genus Pontibacter, designated GY10130T, was isolated from rhizosphere soil of a mangrove plant Rhizophora stylosa collected from Guangxi province, China. Strain GY10130T was Gram-stain negative, positive for oxidase activities, aerobic, short rod-shaped cells without flagella. Growth was observed at 10-40 °C (optimum, 28 °C), pH 6.0-9.0 (optimum, 7.0) and NaCl concentrations of 0-4% (optimum, 1%). Strain GY10130T is closely related to members of the genus Pontibacter, namely P. beigongshangensis CGMCC 1.17104T (97.8%) and P. amylolyticus CGMCC 1.12749T (95.0%), P. humi SWU8T (94.7%), and less than 94.0% with other currently described type strains of Pontibacter. The strain GY10130T showed an ANI value of 80.6% and dDDH value of 23.2% with P. beigongshangensis CGMCC 1.17104T, followed by P. amylolyticus CGMCC 1.12749T with ANI and dDDH values of 72.9 and 13.8%, respectively. Strain GY10130T contains carotenoid-like pigments, but flexirubin-type pigments were absent. The cellular fatty acids (> 10%) consist of summed feature 4 (17:1 iso I/anteiso B) and iso-C15:0. The predominant menaquinone is MK-7. The polar lipids comprise phosphatidylethanolamine, two unidentified glycolipids, two unidentified aminolipids and six unidentified phospholipids. The genome length of strain GY10130T was 6.2 Mbp with a DNA G + C content of 47.1 mol% and 4727 protein-coding genes. The result of polyphasic taxonomic study show strain GY10130T represents one novel species of Pontibacter, Pontibacter qinzhouensis sp. nov., with the type strain GY10130T (=NBRC 113901T = CGMCC 1.16772T).


Assuntos
Rhizophoraceae , Técnicas de Tipagem Bacteriana , Bacteroidetes/genética , China , DNA Bacteriano/genética , Ácidos Graxos/química , Filogenia , RNA Ribossômico 16S/genética , Rizosfera , Análise de Sequência de DNA , Solo , Vitamina K 2/química
18.
Mol Biol Rep ; 49(9): 8381-8390, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35716289

RESUMO

BACKGROUND: Currently, Liver cancer is the fifth most common tumor and the second most important reason for cancer-related death in the world. However, there are still many limitations of the clinical treatment of liver cancer, and new treatment options are clearly needed. Fortunately, studies have shown that L-Selenocysteine has a certain effect on cancer. This study was to investigate the effects of L-Selenocysteine on the inhibition of cell proliferation and the promotion of apoptosis of HepG-2 cells through ROS mediated fine signaling pathway. MATERIALS AND METHODS: CCK-8 assay was applied to evaluating the cytotoxic effect of L-Selenocysteine on HepG-2 cells. Electron microscopy, flow cytometry and Western Blot was utilization in further researching cells signaling pathways. RESULTS: The growth of HepG-2 cells was inhibited by L-selenocysteine ​​treatment in a dose-dependent manner. The cell viability decreased to 52.20%, 43.20% and 30.83% under the treatment of 4, 8, 16 µM L-selenocysteine, respectively. L-Selenocysteine had higher cytotoxicity towards HepG-2 cells than normal cells. L-Selenocysteine can induce the apoptosis of HepG-2 cells by increasing the DNA fragmentation, and activating the Caspase-3. In addition, it was found that the mechanism of the induction to HepG-2 cell apoptosis by L-Selenocysteine was closely related to the overproduction of ROS and promoted apoptosis through the Bcl-2 signaling pathway. CONCLUSIONS: Our data suggest that L-selenocysteine ​​may cause mitochondrial damage and subsequently stimulate ROS production. ROS can damage cellular DNA and mediate the production of Casapase-8, Bid, Bcl-2 and other proteins, affecting downstream signaling pathways, and ultimately induced apoptosis.


Assuntos
Neoplasias Hepáticas , Selenocisteína , Apoptose , Linhagem Celular Tumoral , Humanos , Neoplasias Hepáticas/metabolismo , Potencial da Membrana Mitocondrial , Mitocôndrias/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Espécies Reativas de Oxigênio/metabolismo , Selenocisteína/metabolismo , Selenocisteína/farmacologia , Selenocisteína/uso terapêutico , Transdução de Sinais
19.
Biomed Eng Online ; 21(1): 87, 2022 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-36528597

RESUMO

BACKGROUND: The evaluation of refraction is indispensable in ophthalmic clinics, generally requiring a refractor or retinoscopy under cycloplegia. Retinal fundus photographs (RFPs) supply a wealth of information related to the human eye and might provide a promising approach that is more convenient and objective. Here, we aimed to develop and validate a fusion model-based deep learning system (FMDLS) to identify ocular refraction via RFPs and compare with the cycloplegic refraction. In this population-based comparative study, we retrospectively collected 11,973 RFPs from May 1, 2020 to November 20, 2021. The performance of the regression models for sphere and cylinder was evaluated using mean absolute error (MAE). The accuracy, sensitivity, specificity, area under the receiver operating characteristic curve, and F1-score were used to evaluate the classification model of the cylinder axis. RESULTS: Overall, 7873 RFPs were retained for analysis. For sphere and cylinder, the MAE values between the FMDLS and cycloplegic refraction were 0.50 D and 0.31 D, representing an increase of 29.41% and 26.67%, respectively, when compared with the single models. The correlation coefficients (r) were 0.949 and 0.807, respectively. For axis analysis, the accuracy, specificity, sensitivity, and area under the curve value of the classification model were 0.89, 0.941, 0.882, and 0.814, respectively, and the F1-score was 0.88. CONCLUSIONS: The FMDLS successfully identified the ocular refraction in sphere, cylinder, and axis, and showed good agreement with the cycloplegic refraction. The RFPs can provide not only comprehensive fundus information but also the refractive state of the eye, highlighting their potential clinical value.


Assuntos
Aprendizado Profundo , Retinoscopia , Humanos , Retinoscopia/métodos , Refração Ocular , Midriáticos , Estudos Retrospectivos , Algoritmos
20.
Sensors (Basel) ; 22(17)2022 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-36080990

RESUMO

As a potential air control measure, RF-based surveillance is one of the most commonly used unmanned aerial vehicles (UAV) surveillance methods that exploits specific emitter identification (SEI) technology to identify captured RF signal from ground controllers to UAVs. Recently many SEI algorithms based on deep convolution neural network (DCNN) have emerged. However, there is a lack of the implementation of specific hardware. This paper proposes a high-accuracy and power-efficient hardware accelerator using an algorithm-hardware co-design for UAV surveillance. For the algorithm, we propose a scalable SEI neural network with SNR-aware adaptive precision computation. With SNR awareness and precision reconfiguration, it can adaptively switch between DCNN and binary DCNN to cope with low SNR and high SNR tasks, respectively. In addition, a short-time Fourier transform (STFT) reusing DCNN method is proposed to pre-extract feature of UAV signal. For hardware, we designed a SNR sensing engine, denoising engine, and specialized DCNN engine with hybrid-precision convolution and memory access, aiming at SEI acceleration. Finally, we validate the effectiveness of our design on a FPGA, using a public UAV dataset. Compared with a state-of-the-art algorithm, our method can achieve the highest accuracy of 99.3% and an F1 score of 99.3%. Compared with other hardware designs, our accelerator can achieve the highest power efficiency of 40.12 Gops/W and 96.52 Gops/W with INT16 precision and binary precision.

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