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1.
Proc Natl Acad Sci U S A ; 121(11): e2317430121, 2024 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-38437540

RESUMO

Brown-and-white giant pandas (hereafter brown pandas) are distinct coat color mutants found exclusively in the Qinling Mountains, Shaanxi, China. However, its genetic mechanism has remained unclear since their discovery in 1985. Here, we identified the genetic basis for this coat color variation using a combination of field ecological data, population genomic data, and a CRISPR-Cas9 knockout mouse model. We de novo assembled a long-read-based giant panda genome and resequenced the genomes of 35 giant pandas, including two brown pandas and two family trios associated with a brown panda. We identified a homozygous 25-bp deletion in the first exon of Bace2, a gene encoding amyloid precursor protein cleaving enzyme, as the most likely genetic basis for brown-and-white coat color. This deletion was further validated using PCR and Sanger sequencing of another 192 black giant pandas and CRISPR-Cas9 edited knockout mice. Our investigation revealed that this mutation reduced the number and size of melanosomes of the hairs in knockout mice and possibly in the brown panda, further leading to the hypopigmentation. These findings provide unique insights into the genetic basis of coat color variation in wild animals.


Assuntos
Ursidae , Animais , Camundongos , Ursidae/genética , Peptídeo Hidrolases , Precursor de Proteína beta-Amiloide , Animais Selvagens , Camundongos Knockout
2.
Rev Med Virol ; 34(2): e2522, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38348583

RESUMO

Recently, patients with Mpox breakthrough infection or reinfection were constantly reported. However, the induction, risk factors, and important clinical symptoms of breakthrough infection and reinfection of Mpox virus (MPXV), as well as the factors affecting the effectiveness of Mpox vaccine are not characterized. Herein, a literature review was preformed to summarize the risk factors and important clinical symptoms of patients with Mpox breakthrough infection or reinfection, as well as the factors affecting the effectiveness of smallpox vaccine against Mpox. Results showed that MSM sexual behavior, condomless sexual behavior, multiple sexual partners, close contact, HIV infection, and the presence of comorbidity are important risk factors for Mpox breakthrough infection and reinfection. Genital ulcers, proctitis, and lymphadenopathy are the important clinical symptoms of Mpox breakthrough infection and reinfection. The effectiveness of emergent vaccination of smallpox vaccine for post-exposure of MPXV is associated with smallpox vaccination history, interval between exposure and vaccination, and history of HIV infection. This review provides a better understanding for the risk factors and important clinical symptoms of Mpox breakthrough infection and reinfection, as well as the formulation of Mpox vaccine vaccination strategies.


Assuntos
Infecções por HIV , Mpox , Vacina Antivariólica , Humanos , Reinfecção/epidemiologia , Reinfecção/prevenção & controle , Infecções Irruptivas , Infecções por HIV/complicações , Infecções por HIV/epidemiologia , Antígenos Virais
3.
Plant J ; 116(1): 87-99, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37340958

RESUMO

Nitrogen (N) is a vital major nutrient for rice (Oryza sativa). Rice responds to different applications of N by altering its root morphology, including root elongation. Although ammonium ( NH 4 + ) is the primary source of N for rice, NH 4 + is toxic to rice roots and inhibits root elongation. However, the precise molecular mechanism that NH 4 + -inhibited root elongation of rice is not well understood. Here, we identified a rice T-DNA insert mutant of OsMADS5 with a longer seminal root (SR) under sufficient N conditions. Reverse-transcription quantitative PCR analysis revealed that the expression level of OsMADS5 was increased under NH 4 + compared with NO 3 - supply. Under NH 4 + conditions, knocking out OsMADS5 (cas9) produced a longer SR, phenocopying osmads5, while there was no significant difference in SR length between wild-type and cas9 under NO 3 - supply. Moreover, OsMADS5-overexpression plants displayed the opposite SR phenotype. Further study demonstrated that enhancement of OsMADS5 by NH 4 + supply inhibited rice SR elongation, likely by reducing root meristem activity of root tip, with the involvement of OsCYCB1;1. We also found that OsMADS5 interacted with OsSPL14 and OsSPL17 (OsSPL14/17) to repress their transcriptional activation by attenuating DNA binding ability. Moreover, loss of OsSPL14/17 function in osmads5 eliminated its stimulative effect on SR elongation under NH 4 + conditions, implying OsSPL14/17 may function downstream of OsMADS5 to mediate rice SR elongation under NH 4 + supply. Overall, our results indicate the existence of a novel modulatory pathway in which enhancement of OsMADS5 by NH 4 + supply represses the transcriptional activities of OsSPL14/17 to restrict SR elongation of rice.


Assuntos
Compostos de Amônio , Oryza , Meristema/metabolismo , Oryza/metabolismo , Raízes de Plantas/metabolismo , Compostos de Amônio/metabolismo , Proliferação de Células , Regulação da Expressão Gênica de Plantas
4.
Clin Immunol ; 265: 110300, 2024 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-38950722

RESUMO

A comprehensive analysis of spatial transcriptomics was carried out to better understand the progress of halo nevus. We found that halo nevus was characterized by overactive immune responses, triggered by chemokines and dendritic cells (DCs), T cells, and macrophages. Consequently, we observed abnormal cell death, such as apoptosis and disulfidptosis in halo nevus, some were closely related to immunity. Interestingly, we identified aberrant metabolites such as uridine diphosphate glucose (UDP-G) within the halo nevus. UDP-G, accompanied by the infiltration of DCs and T cells, exhibited correlations with certain forms of cell death. Subsequent experiments confirmed that UDP-G was increased in vitiligo serum and could activate DCs. We also confirmed that oxidative response is an inducer of UDP-G. In summary, the immune response in halo nevus, including DC activation, was accompanied by abnormal cell death and metabolites. Especially, melanocyte-derived UDP-G may play a crucial role in DC activation.

5.
Clin Immunol ; 263: 110199, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38565329

RESUMO

Cell-cell communication is crucial for regulating signaling and cellular function. However, the precise cellular and molecular changes remain poorly understood in skin aging. Based on single-cell and bulk RNA data, we explored the role of cell-cell ligand-receptor interaction in skin aging. We found that the macrophage migration inhibitory factor (MIF)/CD74 ligand-receptor complex was significantly upregulatedin aged skin, showing the predominant paracrine effect of keratinocytes on fibroblasts. Enrichment analysis and in vitro experiment revealed a close association of the activation of the MIF/CD74 with inflammatory pathways and immune response. Mechanistically, MIF/CD74 could significantly inhibit PPARγ protein, which thus significantly increased the degree of fibroblast senescence, and significantly up-regulated the expression of senescence-associated secretory phenotype (SASP) factors and FOS gene. Therefore, our study reveals that MIF/CD74 inhibits the activation of the PPAR signaling pathway, subsequently inducing the production of SASP factors and the upregulation of FOS expression, ultimately accelerating fibroblast senescence.


Assuntos
Antígenos de Diferenciação de Linfócitos B , Fibroblastos , Antígenos de Histocompatibilidade Classe II , Fatores Inibidores da Migração de Macrófagos , Análise de Célula Única , Envelhecimento da Pele , Fatores Inibidores da Migração de Macrófagos/genética , Fatores Inibidores da Migração de Macrófagos/metabolismo , Antígenos de Diferenciação de Linfócitos B/genética , Antígenos de Diferenciação de Linfócitos B/metabolismo , Humanos , Antígenos de Histocompatibilidade Classe II/genética , Antígenos de Histocompatibilidade Classe II/metabolismo , Fibroblastos/metabolismo , Envelhecimento da Pele/genética , Envelhecimento da Pele/fisiologia , Análise de Célula Única/métodos , Transdução de Sinais , Senescência Celular/genética , Oxirredutases Intramoleculares/genética , Oxirredutases Intramoleculares/metabolismo , Análise de Sequência de RNA , Queratinócitos/metabolismo , Queratinócitos/imunologia , PPAR gama/metabolismo , PPAR gama/genética , Pessoa de Meia-Idade , Masculino , Feminino , Pele/metabolismo , Pele/imunologia , Células Cultivadas , Adulto
6.
Environ Sci Technol ; 58(27): 12225-12236, 2024 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-38885124

RESUMO

Nanoscale zerovalent iron synthesized using borohydride (B-NZVI) has been widely applied in environmental remediation in recent decades. However, the contribution of boron in enhancing the inherent reactivity of B-NZVI and its effectiveness in removing hexavalent chromium [Cr(VI)] have not been well recognized and quantified. To the best of our knowledge, herein, a core-shell structure of B-NZVI featuring an Fe-B alloy shell beneath the iron oxide shell is demonstrated for the first time. Alloyed boron can reduce H+, contributing to more than 35.6% of H2 generation during acid digestion of B-NZVIs. In addition, alloyed B provides electrons for Fe3+ reduction during Cr(VI) removal, preventing in situ passivation of the reactive particle surface. Meanwhile, the amorphous oxide shell of B-NZVI exhibits an increased defect density, promoting the release of Fe2+ outside the shell to reduce Cr(VI), forming layer-structured precipitates and intense Fe-O bonds. Consequently, the surface-area-normalized capacity and surface reaction rate of B-NZVI are 6.5 and 6.9 times higher than those of crystalline NZVI, respectively. This study reveals the importance of alloyed B in Cr(VI) removal using B-NZVI and presents a comprehensive approach for investigating electron pathways and mechanisms involved in B-NZVIs for contaminant removal.


Assuntos
Boroidretos , Boro , Ferro , Ferro/química , Boroidretos/química , Boro/química , Cromo/química , Elétrons , Ligas/química
7.
J Adolesc ; 96(1): 196-208, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37908044

RESUMO

BACKGROUND: The adaptation of students to academic challenges in high school is crucial for academic performance. This study proposes the concept of "learning crafting," a previously under-researched area, and investigates its associated variables. METHODS: Using a diary method, we studied 187 Chinese high school students (64% female; Mage = 15.57) over a 9-day period. We examined the effect of daily life events on learning crafting, and considered academic emotions as mediators and regulatory focus as moderators. RESULTS: Hierarchical linear modeling revealed that daily positive events were positively correlated with learning crafting at both within-person and between-person levels. Positive academic emotions served as mediators of this relationship. Furthermore, promotion focus had a positive moderating effect on the relationship between daily positive events and positive academic emotions. Conversely, daily negative events were only negatively correlated with learning crafting at the between-person level, and no additional significant relationships were identified. CONCLUSION: This study elucidated the effect of daily life events on learning crafting, its mediating mechanisms, and conditional factors. These results not only contribute to crafting theory, but also provide theoretical underpinnings for future interventions targeting high school students' learning crafting.


Assuntos
Desempenho Acadêmico , Emoções , Humanos , Feminino , Adolescente , Masculino , Inquéritos e Questionários , Estudantes , Instituições Acadêmicas
8.
J Cell Physiol ; 238(9): 2161-2171, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37417881

RESUMO

Ultraviolet (UV) radiation is the primary exogenous inducer of skin pigmentation, although the mechanism has not been fully elucidated. N6-methyladenosine (m6 A) modification is one of the key epigenetic form of gene regulation that affects multiple biological processes. The aim of this study was to explore the role and underlying mechanisms of m6 A modification in UVB-induced melanogenesis. Low-dose UVB increased global m6 A modification in melanocytes (MCs) and MNT1 melanoma cell line. The GEPIA database predicted that methyltransferase METTL3 is positively correlated with the melanogenic transcription factor MITF in the sun-exposed skin tissues. After METTL3 respectively overexpressed and knocked down in the MNT1, the melanin content and melanogenesis-related genes were significantly upregulated after overexpression of METTL3, especially with UVB irradiation, and downregulated after METTL3 knockdown. METTL3 levels were also higher in melanocytic nevi with high melanin content. METTL3 overexpression and knockdown also altered the protein level of YAP1. SRAMP analysis predicted four high-potential m6 A modification sites on YAP1 mRNA, of which three were confirmed by methylated RNA immunoprecipitation. Inhibition of YAP1 expression can partially reverse melanogenesis induced by overexpression of METTL3. In conclusion, UVB irradiation promotes global m6 A modification in MCs and upregulates METTL3, which increases the expression level of YAP1 through m6 A modification, thereby activating the co-transcription factor TEAD1 and promoting melanogenesis.


Assuntos
Melaninas , Melanócitos , Metiltransferases , Humanos , Melaninas/biossíntese , Melanócitos/metabolismo , Melanócitos/efeitos da radiação , Metiltransferases/genética , Metiltransferases/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Raios Ultravioleta , Linhagem Celular Tumoral
9.
Mol Biol Evol ; 39(6)2022 06 02.
Artigo em Inglês | MEDLINE | ID: mdl-35599233

RESUMO

Incorrect species delimitation will lead to inappropriate conservation decisions, especially for threatened species. The takin (Budorcas taxicolor) is a large artiodactyl endemic to the Himalayan-Hengduan-Qinling Mountains and is well known for its threatened status and peculiar appearance. However, the speciation, intraspecies taxonomy, evolutionary history, and adaptive evolution of this species still remain unclear, which greatly hampers its scientific conservation. Here, we de novo assembled a high-quality chromosome-level genome of takin and resequenced the genomes of 75 wild takins. Phylogenomics revealed that takin was positioned at the root of Caprinae. Population genomics based on the autosome, X chromosome, and Y chromosome SNPs and mitochondrial genomes consistently revealed the existence of two phylogenetic species and recent speciation in takins: the Himalayan takin (B. taxicolor) and the Chinese takin (B. tibetana), with the support of morphological evidence. Two genetically divergent subspecies were identified in both takin species, rejecting three previously proposed taxonomical viewpoints. Furthermore, their distribution boundaries were determined, suggesting that large rivers play important roles in shaping the genetic partition. Compared with the other subspecies, the Qinling subspecies presented the lowest genomic diversity, higher linkage disequilibrium, inbreeding, and genetic load, thus is in urgent need of genetic management and protection. Moreover, coat color gene (PMEL) variation may be responsible for the adaptive coat color difference between the two species following Gloger's rule. Our findings provide novel insights into the recent speciation, local adaptation, scientific conservation of takins, and biogeography of the Himalaya-Hengduan biodiversity hotspot.


Assuntos
Genoma Mitocondrial , Ruminantes , Animais , Espécies em Perigo de Extinção , Especiação Genética , Genômica , Filogenia , Ruminantes/genética
10.
Plant Cell Environ ; 46(11): 3558-3574, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37545348

RESUMO

Adequate distribution of mineral sulphur (S) nutrition to nodules mediated by sulphate transporters is crucial for nitrogen fixation in symbiosis establishment process. However, the molecular mechanisms underlying this process remain largely unknown. In this study, we characterized the function of Early Senescent Nodule 2 (MtESN2), a gene crucial to nitrogen fixation in Medicago truncatula. Mutations in MtESN2 resulted in severe developmental and functional defects including dwarf shoots, early senescent nodules, and lower nitrogenase activity under symbiotic conditions compared to wild-type plants. MtESN2 encodes an M. truncatula sulphate transporter that is expressed only in roots and nodules, with the highest expression levels in the transition zone and nitrogen-fixing zone of nodules. MtESN2 exhibited sulphate transport activity when expressed in yeast. Immunolocalization analysis showed that MtESN2-yellow fluorescent protein fusion protein was localized to the plasma membranes of both uninfected and infected cells of nodules, where it might transport sulphate into both rhizobia-infected and uninfected cells within the nodules. Our results reveal an unreported sulphate transporter that contributes to effective symbiosis and prevents nodule early senescence in M. truncatula.


Assuntos
Medicago truncatula , Fixação de Nitrogênio , Fixação de Nitrogênio/genética , Nódulos Radiculares de Plantas/metabolismo , Medicago truncatula/genética , Medicago truncatula/metabolismo , Transportadores de Sulfato/genética , Transportadores de Sulfato/metabolismo , Simbiose/genética , Sulfatos/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
11.
Exp Dermatol ; 32(4): 310-323, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36394984

RESUMO

The Koebner phenomenon, also known as isomorphic reaction, refers to the development of secondary lesions with the same clinical manifestations and histopathological characteristics as the primary lesions in normal skin after trauma or other stimuli. The triggering factors of Koebner phenomenon include physical trauma, chemical stimulation, mechanical stress, iatrogenic stimulation and pathogenic infection. Vitiligo, psoriasis and lichen planus are considered true Koebner phenomenon. Recent studies have shown that immunological disorders, oxidative stress, defective melanocyte adhesion and growth factor deficiency are the main pathological mechanisms of vitiligo Koebner phenomenon. In psoriasis, triggers may drive skin inflammation to induce a psoriatic phenotype through multiple signalling pathways and thereby cause Koebner phenomenon in susceptible individuals. Significantly, keratinocytes mediate the occurrence of Koebner phenomenon in psoriasis through mechano-induced signalling pathways after sensing mechanical signals and explains the high frequency of psoriasis lesions on the extensor side of the elbow and knee joints. On the contrary, TRPA1-driven mechano-transduction, autoimmunity and actinic damage are the underlying mechanisms of Koebner phenomenon in lichen planus. In this review, we have summarized the current understanding of the characteristics and pathogenesis of Koebner phenomenon.


Assuntos
Dermatite , Líquen Plano , Psoríase , Vitiligo , Humanos , Vitiligo/complicações , Psoríase/patologia
12.
Cell Commun Signal ; 21(1): 332, 2023 11 20.
Artigo em Inglês | MEDLINE | ID: mdl-37986081

RESUMO

Anemia is the most common manifestation in myelodysplastic syndrome (MDS) patients, but the cause of ineffective hematopoiesis is not fully understood. Enucleation is an important event in the maturation process of erythroblasts. According to a series of morphological phenotypes of the pathological development of MDS erythroblasts, we speculate that there may be enucleation disorders. To verify this hypothesis, we cultured MDS bone marrow CD34+ cells in vitro and induced erythroblast development. The results showed that erythroblast enucleation in MDS was significantly lower than that in the normal group, and the rate of enucleation was positively correlated with hemoglobin concentration. Risk stratification of MDS was performed to further analyze the differences in enucleation among the normal group, low-middle risk group and high-risk group. The results showed that the enucleation rate of the high risk group was higher than that of the low-middle risk group but still lower than that of the normal group. Moreover, the expression of pERK and pAKT in MDS erythroblasts in the high risk group was higher than that in the normal group, while the expression of pERK and pAKT in the low-middle risk group was lower than that in the normal group. Furthermore, the enucleation of MDS was positively correlated with the phosphorylation degree of ERK and AKT. In conclusion, this study reveals that the enucleation of erythroblasts is one of the possible causes of anemia in MDS. Video Abstract.


Assuntos
Anemia , Síndromes Mielodisplásicas , Humanos , Eritroblastos/metabolismo , Eritroblastos/patologia , Síndromes Mielodisplásicas/complicações , Síndromes Mielodisplásicas/metabolismo , Anemia/complicações , Anemia/metabolismo , Anemia/patologia , Fatores de Risco , Células da Medula Óssea/patologia
13.
J Fish Dis ; 46(2): 99-112, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36263741

RESUMO

The Streptomyces lateritius Z1-26 was isolated from soil samples which showed broad-spectrum antibacterial activity against a broad range of fish pathogens. The In Vivo Imaging System (IVIS) monitored that strain Z1-26 could survive and colonize in the gills and abdomen of crucian carp. The effects of dietary supplementation with strain Z1-26 were evaluated with respect to the growth performance, antioxidant capacity, and immune response of crucian carp. The results showed that the Z1-26-fed fish had a significantly higher growth rate than the fish fed the control diet. The immune and antioxidant parameters revealed that the non-specific immune indicators (AKP, SOD, and LZM) of the serum, the expression of immune-related genes (IgM, C3, and LZM), and antioxidant-related genes (Nrf2 and Keap1) of the immune organs were significantly increased, whereas the expression of pro-inflammatory factors (IL-1ß, IL-8, and TNF-α) of the immune organs was significantly down-regulated in crucian carp fed strain Z1-26 compared with fish fed a control diet. Moreover, fish fed with Z1-26 supplemented diets showed a significantly improved survival rate after Aeromonas hydrophila infection. In addition, the whole genome analysis showed that strain Z1-26 possesses 28 gene clusters, including 6 polyketide synthetase (PKS), 4 non-ribosomal peptide-synthetase (NRPS), 1 bacteriocin, and 1 lantipeptide. In summary, these results indicated that strain Z1-26 could improve the growth performance and disease resistance in crucian carp, and has the potential to be developed as a candidate probiotics for the control of bacterial diseases in aquaculture.


Assuntos
Carpas , Doenças dos Peixes , Infecções por Bactérias Gram-Negativas , Animais , Carpa Dourada/genética , Carpas/metabolismo , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Antioxidantes/metabolismo , Doenças dos Peixes/microbiologia , Fator 2 Relacionado a NF-E2/metabolismo , Dieta , Antibacterianos/farmacologia , Aeromonas hydrophila/fisiologia , Proteínas de Peixes/genética , Ração Animal/análise
14.
Mol Biol Evol ; 38(2): 531-544, 2021 01 23.
Artigo em Inglês | MEDLINE | ID: mdl-32960966

RESUMO

Antagonistic coevolution between host and parasite drives species evolution. However, most of the studies only focus on parasitism adaptation and do not explore the coevolution mechanisms from the perspective of both host and parasite. Here, through the de novo sequencing and assembly of the genomes of giant panda roundworm, red panda roundworm, and lion roundworm parasitic on tiger, we investigated the genomic mechanisms of coevolution between nonmodel mammals and their parasitic roundworms and those of roundworm parasitism in general. The genome-wide phylogeny revealed that these parasitic roundworms have not phylogenetically coevolved with their hosts. The CTSZ and prolyl 4-hydroxylase subunit beta (P4HB) immunoregulatory proteins played a central role in protein interaction between mammals and parasitic roundworms. The gene tree comparison identified that seven pairs of interactive proteins had consistent phylogenetic topology, suggesting their coevolution during host-parasite interaction. These coevolutionary proteins were particularly relevant to immune response. In addition, we found that the roundworms of both pandas exhibited higher proportions of metallopeptidase genes, and some positively selected genes were highly related to their larvae's fast development. Our findings provide novel insights into the genetic mechanisms of coevolution between nonmodel mammals and parasites and offer the valuable genomic resources for scientific ascariasis prevention in both pandas.


Assuntos
Ascaridoidea/genética , Coevolução Biológica , Genoma Helmíntico , Interações Hospedeiro-Parasita/genética , Tigres/parasitologia , Ursidae/parasitologia , Animais , Filogenia , Mapas de Interação de Proteínas , Seleção Genética , Tigres/genética , Tigres/metabolismo , Ursidae/genética , Ursidae/metabolismo
15.
Microb Pathog ; 169: 105646, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35716927

RESUMO

Aeromonas veronii AvX005 is a pathogenic bacterium with high toxicity to grass carp (Ctenopharyngodon idellus). The expression levels of g-type (goose-type lysozyme, Lys-g) and c-type lysozyme (chicken-type lysozyme, Lys-c) in the spleen of grass carp infected with AvX005 were significantly increased by approximately 4.5 times and 27 times, respectively. The recombinant proteins rLys-g and rLys-c produced in a recombinant expression system of Escherichia coli showed significant antibacterial activity against the pathogenic bacteria AvX005. A challenge test was conducted after rLys-g and rLys-c were expressed in grass carp L8824 liver cells, and compared with the survival rate of the control cells (46.3%), the survival rate of the experimental cells (77.6% for rLys-g and 68.6% for rLys-c) was significantly increased. Grass carp were infected with AvX005 on the second day after delivering pcDNA3.1-lys-g and pcDNA-lys-c with the Quil A/cholesterol/DDA/Carbopol (QCDC) adjuvant, and both pcDNA3.1-lys-g and pcDNA-lys-c provided 70% relative protection for grass carp. The activity of lysozyme and alkaline phosphatase in the serum of grass carp was significantly increased after injection of DNA. The expression of the immune factors IgM, C3 and IL8 in the kidney was upregulated to varying degrees for pcDNA3.1-lys-g and immune factors C3 and IgM was upregulated for pcDNA-lys-c. The results indicated that pcDNA3.1-lys-g and pcDNA-lys-c may be used as immunostimulants to protect grass carp from the pathogenic bacterium AvX005.


Assuntos
Carpas , Doenças dos Peixes , Infecções por Bactérias Gram-Negativas , Resinas Acrílicas , Adjuvantes Imunológicos/farmacologia , Aeromonas hydrophila/fisiologia , Aeromonas veronii , Animais , Carpas/metabolismo , Colesterol , Doenças dos Peixes/microbiologia , Imunidade Inata , Imunoglobulina M , Muramidase/genética , Muramidase/farmacologia , Saponinas de Quilaia
16.
J Nat Prod ; 85(4): 838-845, 2022 04 22.
Artigo em Inglês | MEDLINE | ID: mdl-35290062

RESUMO

Hyperpigmented skin diseases such as melasma, freckles, and melanosis usually mar the appearance of patients. Traditional herbal medicines are highly accepted in inhibiting skin pigmentation, with advantages of high efficiency, low cost, and low side effects. Selaginellin (SEL), one of the active compounds of selaginella, has been proved to be exhibit antineoplastic, antioxidant, antisenescence, and antiapoptosis activities. In this study, we found that SEL can inhibit melanogenesis in vitro and in vivo. A mechanism study found that SEL inhibits melanogenesis through inhibiting the mitogen-activated protein kinase (MAPK) signaling pathway, then down-regulating the expression of microphthalmia-associated transcription factor (MITF) and downstream genes tyrosinase (TYR) and tyrosinase-related protein 2 (TYRP2). UVB-activated paracrine function of fibroblasts and keratinocytes promotes melanogenesis of melanocytes. Interestingly, SEL antagonizes UVB-activated paracrine function of fibroblasts and keratinocytes. These findings indicate that SEL can be a potential whitening compound to inhibit melanogenesis.


Assuntos
Melaninas , Proteínas Quinases Ativadas por Mitógeno , Humanos , Melanócitos , Fator de Transcrição Associado à Microftalmia/genética , Fator de Transcrição Associado à Microftalmia/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Monofenol Mono-Oxigenase/metabolismo , Transdução de Sinais
17.
Appl Microbiol Biotechnol ; 106(2): 647-661, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-35019997

RESUMO

Filamentous fungi are extensively used as an important expression host for the production of a variety of essential industrial proteins. They have significant promise as an expression system for protein synthesis due to their inherent superior secretory capabilities. The purpose of this study was to develop a novel expression system by utilizing a Penicillium oxalicum strain that possesses a high capacity for protein secretion. The expression of glycoside hydrolases in P. oxalicum was evaluated in a cleaner extracellular background where the formation of two major amylases was inhibited. Four glycoside hydrolases (CBHI, Amy15B, BGL1, and Cel12A) were expressed under the highly constitutive promoter PubiD. It was found that the proteins exhibited high purity in the culture supernatant after cultivation with starch. Two inducible promoters, Pamy15A and PempA, under the activation of the transcription factor AmyR were used as elements in the construction of versatile vectors. When using the cellobiohydrolase CBHI as the extracellular quantitative reporter, the empA promoter screened from the AmyR-overexpressing strain was shown to be superior to the amy15A promoter based on RNA-sequencing data. Therefore, we designed an expression system consisting of a cleaner background host strain and an adjustable promoter. This system enables rapid and high-throughput evaluation of glycoside hydrolases from filamentous fungi.Key points• A new protein expression system derived from Penicillium oxalicum has been developed.• The expression platform is capable of secreting recombinant proteins with high purity.• The adjustable promoter may allow for further optimization of recombinant protein synthesis.


Assuntos
Proteínas Fúngicas , Penicillium , Amilases/genética , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Fungos/metabolismo , Glicosídeo Hidrolases/genética , Penicillium/genética , Penicillium/metabolismo , Regiões Promotoras Genéticas
18.
Appl Microbiol Biotechnol ; 106(23): 7857-7866, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36326838

RESUMO

Xenorhabdus can produce a large number of secondary metabolites with insecticidal, bacteriostatic, and antitumor activities. Efficient gene editing tools will undoubtedly facilitate the functional genomics research and bioprospecting in Xenorhabdus. In this study, BlastP analysis using the amino acid sequences of Redαß or RecET recombinases as queries resulted in the identification of an operon (XBJ1_operon 0213) containing RecET-like recombinases encoding genes from the genome of Xenorhabdus bovienii strain SS-2004. Three proteins encoded by this operon was indispensable for full activity of recombineering, namely XBJ1-1173 (RecE-like protein), XBJ1-1172 (RecT-like protein), and XBJ1-1171 (single-strand annealing protein). Using this newly developed recombineering system, a gene cluster responsible for biosynthesis of a novel secondary metabolite (Min16) was identified from X. stockiae HN_xs01 strain. Min16 which exhibited antibacterial and cytotoxic activities was determined to be a cyclopeptide composed of Acyl-Phe-Thr-Phe-Pro-Pro-Leu-Val by using high-resolution mass spectrometry and nuclear magnetic resonance analysis, and was designated as changshamycin. This host-specific recombineering system was proven to be effective for gene editing in Xenorhabdus, allowing for efficient discovery of novel natural products with attractive bioactivities. KEY POINTS: • Screening and identification of efficient gene editing tools from Xenorhabdus • Optimization of the Xenorhabdus electroporation parameters • Discovery of a novel cyclopeptide compound with multiple biological activities.


Assuntos
Produtos Biológicos , Xenorhabdus , Xenorhabdus/genética , Recombinases/genética , Recombinases/metabolismo , Produtos Biológicos/metabolismo , Óperon , Peptídeos Cíclicos/metabolismo
19.
Appl Microbiol Biotechnol ; 106(8): 3081-3091, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35376972

RESUMO

PII signal transduction proteins are widely found in bacteria and plant chloroplast, and play a central role in nitrogen metabolism regulation, which interact with many key proteins in metabolic pathways to regulate carbon/nitrogen balance by sensing changes in concentrations of cell-mediated indicators such as α-ketoglutarate. In this study, the knockout strain Saccharopolyspora pogona-ΔpII and overexpression strain S. pogona-pII were constructed using CRISPR/Cas9 technology and the shuttle vector POJ260, respectively, to investigate the effects on the growth and secondary metabolite biosynthesis of S. pogona. Growth curve, electron microscopy, and spore germination experiments were performed, and it was found that the deletion of the pII gene inhibited the growth to a certain extent in the mutant. HPLC analysis showed that the yield of butenyl-spinosyn in the S. pogona-pII strain increased to 245% than that in the wild-type strain while that in S. pogona-ΔpII decreased by approximately 51%. This result showed that the pII gene can promote the growth and butenyl-spinosyn biosynthesis of S. pogona. This research first investigated PII nitrogen metabolism regulators in S. pogona, providing significant scientific evidence and a research basis for elucidating the mechanism by which these factors regulate the growth of S. pogona, optimizing the synthesis network of butenyl-spinosyn and constructing a strain with a high butenyl-spinosyn yield. KEY POINTS: • pII key nitrogen regulatory gene deletion can inhibit the growth and development of S. pogona. • Overexpressed pII gene can significantly promote the butenyl-spinosyn biosynthesis. • pII gene can affect the amino acid circulation and the accumulation of butenyl-spinosyn precursors in S. pogona.


Assuntos
Nitrogênio , Saccharopolyspora , Proteínas de Bactérias/genética , Genes Reguladores , Macrolídeos/metabolismo , Nitrogênio/metabolismo , Saccharopolyspora/metabolismo
20.
Angew Chem Int Ed Engl ; 61(37): e202208587, 2022 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-35791044

RESUMO

Interfacial photo-vapor conversion has been suggested as a cost-effective and sustainable technology for seawater desalination. However, the conversion performance was still limited by some drawbacks, like salt accumulation and poor mechanical stability. Herein, a scalable MoS2 -based porous hydrogel (SMoS2 -PH) with good mechanical stability and salt resistance was successfully constructed through a crosslinking foaming polymerization method. With the high porosity (92.63 %), the SMoS2 -PH performed an impressive evaporation rate of 3.297 kg m-2 h-1 and photothermal conversion efficiency of 93.4 % under 1-sun illumination. Most importantly, the SMoS2 -PH could maintain high and stable photothermal properties for 15 days on the surface of seawater. We believe that the excellent salt resistance, the high photothermal conversion efficiency, the ease of scale preparation method and the available commercial MoS2 make the SMoS2 -PH a promising device for full-scale seawater desalination.

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