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1.
J Bacteriol ; : e0003324, 2024 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-38899896

RESUMO

Listeria monocytogenes is a foodborne bacterial pathogen that causes listeriosis. Positive regulatory factor A (PrfA) is a pleiotropic master activator of virulence genes of L. monocytogenes that becomes active upon the entry of the bacterium into the cytosol of infected cells. L. monocytogenes can survive and multiply at low temperatures; this is accomplished through the maintenance of appropriate membrane fluidity via branched-chain fatty acid (BCFA) synthesis. Branched-chain α-keto acid dehydrogenase (BKD), which is composed of four polypeptides encoded by lpd, bkdA1, bkdA2, and bkdB, is known to play a vital role in BCFA biosynthesis. Here, we constructed BKD-deficient Listeria strains by in-frame deletion of lpd, bkdA1, bkdA2, and bkdB genes. To determine the role in in vivo and in vitro, mouse model challenges, plaque assay in murine L2 fibroblast, and intracellular replication in J744A.1 macrophage were conducted. BKD-deficient strains exhibited defects in BCFA composition, virulence, and PrfA-regulon function within the host cells. Transcriptomics analysis revealed that the transcript level of the PrfA-regulon was lower in ΔbkdA1 strain than those in the wild-type. This study demonstrates that L. monocytogenes strains lacking BKD complex components were defective in PrfA-regulon function, and full activation of wild-type prfA may not occur within host cells in the absence of BKD. Further study will investigate the consequences of BKD deletion on PrfA function through altering BCFA catabolism.IMPORTANCEListeria monocytogenes is the causative agent of listeriosis, a disease with a high mortality rate. In this study, we have shown that the deletion of BKD can impact the function of PrfA and the PrfA-regulon. The production of virulence proteins within host cells is necessary for L. monocytogenes to promote its intracellular survival and is likely dependent on membrane integrity. We thus report a link between L. monocytogenes membrane integrity and the function of PrfA. This knowledge will increase our understanding of L. monocytogenes pathogenesis, which may provide insight into the development of antimicrobial agents.

2.
Biochem Biophys Res Commun ; 727: 150309, 2024 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-38936224

RESUMO

Versican is a large chondroitin sulfate proteoglycan in the extracellular matrix. It plays a pivotal role in the formation of the provisional matrix. S100a4, previously known as fibroblast-specific protein, functions as a calcium channel-binding protein. To investigate the role of versican expressed in fibroblasts, we generated conditional knockout mice in which versican expression is deleted in cells expressing S100a4. We found that S100a4 is expressed in adipose tissues, and these mice exhibit obesity under a normal diet, which becomes apparent as early as five months. The white adipose tissues of these mice exhibited decreased expression levels of S100a4 and versican and hypertrophy of adipocytes. qRT-PCR showed a reduced level of UCP1 in their white adipose tissues, indicating that the basic energy metabolism is diminished. These results suggest that versican in adipose tissues maintains the homeostasis of adipose tissues and regulates energy metabolism.

3.
Ann Rheum Dis ; 2024 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-38902010

RESUMO

OBJECTIVES: Autoantibodies targeting intracellular proteins are common in various autoimmune diseases. In the context of myositis, the pathologic significance of these autoantibodies has been questioned due to the assumption that autoantibodies cannot enter living muscle cells. This study aims to investigate the validity of this assumption. METHODS: Confocal immunofluorescence microscopy was employed to localise antibodies and other proteins of interest in myositis muscle biopsies. Bulk RNA sequencing was used to examine the transcriptomic profiles of 669 samples, including those from patients with myositis, disease controls and healthy controls. Additionally, antibodies from myositis patients were introduced into cultured myoblasts through electroporation, and their transcriptomic profiles were analysed using RNA sequencing. RESULTS: In patients with myositis autoantibodies, antibodies accumulated inside myofibres in the same subcellular compartment as the autoantigen. Bulk RNA sequencing revealed that muscle biopsies from patients with autoantibodies targeting transcriptional regulators exhibited transcriptomic patterns consistent with dysfunction of the autoantigen. For instance, in muscle biopsies from patients with anti-PM/Scl autoantibodies recognising components of the nuclear RNA exosome complex, an accumulation of divergent transcripts and long non-coding RNAs was observed; these RNA forms are typically degraded by the nuclear RNA exosome complex. Introducing patient antibodies into cultured muscle cells recapitulated the transcriptomic effects observed in human disease. Further supporting evidence suggested that myositis autoantibodies recognising other autoantigens may also disrupt the function of their targets. CONCLUSIONS: This study demonstrates that, in myositis, autoantibodies are internalised into living cells, causing biological effects consistent with the disrupted function of their autoantigen.

4.
Proc Natl Acad Sci U S A ; 115(50): 12793-12798, 2018 12 11.
Artigo em Inglês | MEDLINE | ID: mdl-30487218

RESUMO

DNA damage tolerance (DDT) releases replication blockage caused by damaged nucleotides on template strands employing two alternative pathways, error-prone translesion DNA synthesis (TLS) and error-free template switch (TS). Lys164 of proliferating cell nuclear antigen (PCNA) is SUMOylated during the physiological cell cycle. To explore the role for SUMOylation of PCNA in DDT, we characterized chicken DT40 and human TK6 B cells deficient in the PIAS1 and PIAS4 small ubiquitin-like modifier (SUMO) E3 ligases. DT40 cells have a unique advantage in the phenotypic analysis of DDT as they continuously diversify their immunoglobulin (Ig) variable genes by TLS and TS [Ig gene conversion (GC)], both relieving replication blocks at abasic sites without accompanying by DNA breakage. Remarkably, PIAS1-/-/PIAS4-/- cells displayed a multifold decrease in SUMOylation of PCNA at Lys164 and over a 90% decrease in the rate of TS. Likewise, PIAS1-/-/PIAS4-/- TK6 cells showed a shift of DDT from TS to TLS at a chemosynthetic UV lesion inserted into the genomic DNA. The PCNAK164R/K164R mutation caused a ∼90% decrease in the rate of Ig GC and no additional impact on PIAS1-/-/PIAS4-/- cells. This epistatic relationship between the PCNAK164R/K164R and the PIAS1-/-/PIAS4-/- mutations suggests that PIAS1 and PIAS4 promote TS mainly through SUMOylation of PCNA at Lys164. This idea is further supported by the data that overexpression of a PCNA-SUMO1 chimeric protein restores defects in TS in PIAS1-/-/PIAS4-/- cells. In conclusion, SUMOylation of PCNA at Lys164 promoted by PIAS1 and PIAS4 ensures the error-free release of replication blockage during physiological DNA replication in metazoan cells.


Assuntos
Linfócitos B/metabolismo , Galinhas/genética , Proteínas de Ligação a Poli-ADP-Ribose/genética , Antígeno Nuclear de Célula em Proliferação/genética , Proteínas Inibidoras de STAT Ativados/genética , Sumoilação/genética , Animais , Ciclo Celular/genética , Linhagem Celular , Dano ao DNA/genética , Reparo do DNA/genética , Replicação do DNA/genética , Conversão Gênica/genética , Genes de Imunoglobulinas/genética , Humanos , Região Variável de Imunoglobulina/genética , Ubiquitina-Proteína Ligases/genética
5.
FASEB J ; 32(7): 3583-3596, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29452569

RESUMO

Although a deficiency of surfactant protein B (SFTPB) has been associated with lung injury, SFTPB expression has not yet been linked with nicotinamide phosphoribosyltransferase (NAMPT), a potential biomarker of acute lung injury (ALI). The effects of Nampt in the pulmonary epithelial cell on both SFTPB expression and lung inflammation were investigated in a LPS-induced ALI mouse model. Pulmonary epithelial cell-specific knockdown of Nampt gene expression, achieved by the crossing of Nampt gene exon 2 floxed mice with mice expressing epithelial-specific transgene Cre or by the use of epithelial-specific expression of anti-Nampt antibody cDNA, significantly attenuated LPS-induced ALI. Knockdown of Nampt expression was accompanied by lower levels of bronchoalveolar lavage (BAL) neutrophil infiltrates, total protein and TNF-α levels, as well as lower lung injury scores. Notably, Nampt knockdown was also associated with significantly increased BAL SFTPB levels relative to the wild-type control mice. Down-regulation of NAMPT increased the expression of SFTPB and rescued TNF-α-induced inhibition of SFTPB, whereas overexpression of NAMPT inhibited SFTPB expression in both H441 and A549 cells. Inhibition of NAMPT up-regulated SFTPB expression by enhancing histone acetylation to increase its transcription. Additional data indicated that these effects were mainly mediated by NAMPT nonenzymatic function via the JNK pathway. This study shows that pulmonary epithelial cell-specific knockdown of NAMPT expression attenuated ALI, in part, via up-regulation of SFTPB expression. Thus, epithelial cell-specific knockdown of Nampt may be a potential new and viable therapeutic modality to ALI.-Bi, G., Wu, L., Huang, P., Islam, S., Heruth, D. P., Zhang, L. Q., Li, D.-Y., Sampath, V., Huang, W., Simon, B. A., Easley, R. B., Ye, S. Q. Up-regulation of SFTPB expression and attenuation of acute lung injury by pulmonary epithelial cell-specific NAMPT knockdown.


Assuntos
Lesão Pulmonar Aguda/metabolismo , Células Epiteliais Alveolares/metabolismo , Citocinas/genética , Nicotinamida Fosforribosiltransferase/genética , Surfactantes Pulmonares/metabolismo , Lesão Pulmonar Aguda/genética , Animais , Linhagem Celular Tumoral , Citocinas/metabolismo , Histonas/metabolismo , Humanos , MAP Quinase Quinase 4/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Nicotinamida Fosforribosiltransferase/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Regulação para Cima
6.
Nucleic Acids Res ; 43(13): 6359-72, 2015 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-26089390

RESUMO

Smarcal1 is a SWI/SNF-family protein with an ATPase domain involved in DNA-annealing activities and a binding site for the RPA single-strand-DNA-binding protein. Although the role played by Smarcal1 in the maintenance of replication forks has been established, it remains unknown whether Smarcal1 contributes to genomic DNA maintenance outside of the S phase. We disrupted the SMARCAL1 gene in both the chicken DT40 and the human TK6 B cell lines. The resulting SMARCAL1(-/-) clones exhibited sensitivity to chemotherapeutic topoisomerase 2 inhibitors, just as nonhomologous end-joining (NHEJ) null-deficient cells do. SMARCAL1(-/-) cells also exhibited an increase in radiosensitivity in the G1 phase. Moreover, the loss of Smarcal1 in NHEJ null-deficient cells does not further increase their radiosensitivity. These results demonstrate that Smarcal1 is required for efficient NHEJ-mediated DSB repair. Both inactivation of the ATPase domain and deletion of the RPA-binding site cause the same phenotype as does null-mutation of Smarcal1, suggesting that Smarcal1 enhances NHEJ, presumably by interacting with RPA at unwound single-strand sequences and then facilitating annealing at DSB ends. SMARCAL1(-/-)cells showed a poor accumulation of Ku70/DNA-PKcs and XRCC4 at DNA-damage sites. We propose that Smarcal1 maintains the duplex status of DSBs to ensure proper recruitment of NHEJ factors to DSB sites.


Assuntos
Quebras de DNA de Cadeia Dupla , Reparo do DNA por Junção de Extremidades , DNA Helicases/fisiologia , Animais , Camptotecina/toxicidade , Linhagem Celular , Galinhas , Dano ao DNA , DNA Helicases/química , DNA Helicases/genética , Proteínas de Ligação a DNA/metabolismo , Fase G1/genética , Deleção de Genes , Humanos , Estrutura Terciária de Proteína
7.
Genes Cells ; 19(10): 743-54, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25205350

RESUMO

RING finger protein 4 (RNF4) represents a subclass of ubiquitin ligases that target proteins modified by the small ubiquitin-like modifier (SUMO) for ubiquitin-mediated degradation. We disrupted the RNF4 gene in chicken DT40 cells and found that the resulting RNF4(-/-) cells gradually lost proliferation capability. Strikingly, this compromised proliferation was associated with an unprecedented cellular effect: the gradual decrease in the number of intact chromosomes. In the 6 weeks after gene targeting, there was a 25% reduction in the DNA content of the RNF4(-/-) cells. Regarding trisomic chromosome 2, 60% of the RNF4(-/-) cells lost one homologue, suggesting that DNA loss was mediated by whole chromosome loss. To determine the cause of this chromosome loss, we examined cell-cycle checkpoint pathways. RNF4(-/-) cells showed a partial defect in the spindle assembly checkpoint, premature dissociation of sister chromatids, and a marked increase in the number of lagging chromosomes at anaphase. Thus, combined defects in SAC and sister chromatid cohesion may result in increased lagging chromosomes, leading to chromosome loss without accompanying chromosome gain in RNF4(-/-) cells. We therefore propose that RNF4 plays a novel role in preventing the loss of intact chromosomes and ensures the maintenance of chromosome integrity.


Assuntos
Aberrações Cromossômicas , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Anáfase , Animais , Linhagem Celular , Proliferação de Células , Sobrevivência Celular/genética , Galinhas , Segregação de Cromossomos , Dano ao DNA , Replicação do DNA , Pontos de Checagem da Fase M do Ciclo Celular , Mutação , Ubiquitina-Proteína Ligases/genética
8.
Arch Toxicol ; 88(1): 145-60, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23963510

RESUMO

Although carbon nanotubes (CNTs) are promising nanomaterials, their potential carcinogenicity is a major concern. We previously established a genetic method of analyzing genotoxicity of chemical compounds, where we evaluated their cytotoxic effect on the DT40 lymphoid cell line comparing DNA-repair-deficient isogenic clones with parental wild-type cells. However, application of our DT40 system for the cytotoxic and genotoxic evaluation of nanomaterials seemed to be difficult, because DT40 cells only poorly internalized nanoparticles. To solve this problem, we have constructed a chimeric gene encoding a trans-membrane receptor consisting of the 5' region of the transferrin receptor (TR) gene (to facilitate internalization of nanoparticles) and the 3' region of the macrophage receptor with collagenous structure (MARCO) gene (which is a receptor for environmental particles). We expressed the resulting MARCO-TR chimeric receptor on DNA-repair-proficient wild-type cells and mutants deficient in base excision repair (FEN1 (-/-)) and translesion DNA synthesis (REV3 (-/-)). We demonstrated that the chimera mediates uptake of particles such as fluorescence-tagged polystyrene particles and multi-walled carbon nanotubes (MWCNTs), with very poor uptake of those particles by DT40 cells not expressing the chimera. MWCNTs were cytotoxic and this effect was greater in FEN1 (-/-)and REV3 (-/-) cells than in wild-type cells. Furthermore, MWCNTs induced greater oxidative damage (measured as 8-OH-dG formation) and a larger number of mitotic chromosomal aberrations in repair-deficient cells compared to repair-proficient cells. Taken together, our novel assay system using the chimeric receptor-expressing DT40 cells provides a sensitive method to screen for genotoxicity of CNTs and possibly other nanomaterials.


Assuntos
Linfócitos B/efeitos dos fármacos , Testes de Mutagenicidade/métodos , Nanotubos de Carbono/toxicidade , Receptores Imunológicos/metabolismo , 8-Hidroxi-2'-Desoxiguanosina , Animais , Linhagem Celular/efeitos dos fármacos , Galinhas , Aberrações Cromossômicas , Reparo do DNA/efeitos dos fármacos , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , Poliestirenos/farmacocinética , Receptores Imunológicos/genética , Receptores da Transferrina/genética , Receptores da Transferrina/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo
9.
Front Microbiol ; 15: 1403694, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38881664

RESUMO

Listeria monocytogenes causes listeriosis, an infectious and potentially fatal disease of animals and humans. A diverse network of transcriptional regulators, including LysR-type catabolite control protein C (CcpC), is critical for the survival of L. monocytogenes and its ability to transition into the host environment. In this study, we explored the physiological and genetic consequences of deleting ccpC and the effects of such deletion on the ability of L. monocytogenes to cause disease. We found that ccpC deletion did not impact hemolytic activity, whereas it resulted in significant reductions in phospholipase activities. Western blotting revealed that the ΔccpC strain produced significantly reduced levels of the cholesterol-dependent cytolysin LLO relative to the wildtype F2365 strain. However, the ΔccpC mutant displayed no significant intracellular growth defect in macrophages. Furthermore, ΔccpC strain exhibited reduction in plaque numbers in fibroblasts compared to F2365, but plaque size was not significantly affected by ccpC deletion. In a murine model system, the ΔccpC strain exhibited a significantly reduced bacterial burden in the liver and spleen compared to the wildtype F2365 strain. Interestingly, the deletion of this gene also enhanced the survival of L. monocytogenes under conditions of H2O2-induced oxidative stress. Transcriptomic analyses performed under H2O2-induced oxidative stress conditions revealed that DNA repair, cellular responses to DNA damage and stress, metalloregulatory proteins, and genes involved in the biosynthesis of peptidoglycan and teichoic acids were significantly induced in the ccpC deletion strain relative to F2365. In contrast, genes encoding internalin, 1-phosphatidylinositol phosphodiesterase, and genes associated with sugar-specific phosphotransferase system components, porphyrin, branched-chain amino acids, and pentose phosphate pathway were significantly downregulated in the ccpC deletion strain relative to F2365. This finding highlights CcpC as a key factor that regulates L. monocytogenes physiology and responses to oxidative stress by controlling the expression of important metabolic pathways.

10.
medRxiv ; 2024 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-38313303

RESUMO

Objectives: Myositis is a heterogeneous family of autoimmune muscle diseases. As myositis autoantibodies recognize intracellular proteins, their role in disease pathogenesis has been unclear. This study aimed to determine whether myositis autoantibodies reach their autoantigen targets within muscle cells and disrupt the normal function of these proteins. Methods: Confocal immunofluorescence microscopy was used to localize antibodies and other proteins of interest in myositis muscle biopsies. Bulk RNA sequencing was used to study the transcriptomic profiles of 668 samples from patients with myositis, disease controls, and healthy controls. Antibodies from myositis patients were introduced into cultured myoblasts by electroporation and the transcriptomic profiles of the treated myoblasts were studied by bulk RNA sequencing. Results: In patients with myositis autoantibodies, antibodies accumulated inside myofibers in the same subcellular compartment as the autoantigen. Each autoantibody was associated with effects consistent with dysfunction of its autoantigen, such as the derepression of genes normally repressed by Mi2/NuRD in patients with anti-Mi2 autoantibodies, the accumulation of RNAs degraded by the nuclear RNA exosome complex in patients with anti-PM/Scl autoantibodies targeting this complex, and the accumulation of lipids within myofibers of anti-HMGCR-positive patients. Internalization of patient immunoglobulin into cultured myoblasts recapitulated the transcriptomic phenotypes observed in human disease, including the derepression of Mi2/NuRD-regulated genes in anti-Mi2-positive dermatomyositis and the increased expression of genes normally degraded by the nuclear RNA exosome complex in anti-PM/Scl-positive myositis. Conclusions: In myositis, autoantibodies are internalized into muscle fibers, disrupt the biological function of their autoantigen, and mediate the pathophysiology of the disease.

11.
J Biol Chem ; 287(15): 11656-64, 2012 Apr 06.
Artigo em Inglês | MEDLINE | ID: mdl-22334704

RESUMO

T lymphocytes circulate between the blood, tissues, and lymph. These T cells carry out immune functions, using the C-C chemokine receptor 7 (CCR7) and its cognate ligands, CCL19 and CCL21, to enter and travel through the lymph nodes. Distinct roles for each ligand in regulating T lymphocyte trafficking have remained elusive. We report that in the human T cell line HuT78 and in primary murine T lymphocytes, signaling from CCR7/CCL19 leads to increased expression and phosphorylation of extracellular signal-regulated kinase 5 (ERK5) within eight hours of stimulation. Within 48-72 h we observed peak levels of endothelial differentiation gene 1 (EDG-1), which mediates the egress of T lymphocytes from lymph nodes. The increased expression of EDG-1 was preceded by up-regulation of its transcription factor, Krüppel-like factor 2 (KLF-2). To determine the cellular effect of disrupting ERK5 signaling from CCR7, we examined the migration of ERK5(flox/flox)/Lck-Cre murine T cells to EDG-1 ligands. While CCL19-stimulated ERK5(flox/flox) naïve T cells showed increased migration to EDG-1 ligands at 48 h, the migration of ERK5(flox/flox)/Lck-Cre T cells remained at a basal level. Accordingly, we define a novel signaling pathway that controls EDG-1 up-regulation following stimulation of T cells by CCR7/CCL19. This is the first report to link the two signaling events that control migration through the lymph nodes: CCR7 mediates entry into the lymph nodes and EDG-1 signaling controls their subsequent exit.


Assuntos
Quimiocina CCL19/fisiologia , Regulação da Expressão Gênica , Receptores CCR7/fisiologia , Receptores de Lisoesfingolipídeo/metabolismo , Linfócitos T/metabolismo , Transporte Ativo do Núcleo Celular , Animais , Linhagem Celular , Quimiocina CCL19/genética , Quimiocina CCL19/metabolismo , Quimiocina CCL21/fisiologia , Quimiotaxia , Células Dendríticas/metabolismo , Expressão Gênica , Humanos , Lisofosfolipídeos/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Proteína Quinase 7 Ativada por Mitógeno/genética , Proteína Quinase 7 Ativada por Mitógeno/metabolismo , Fosforilação , Receptores CCR7/metabolismo , Receptores de Lisoesfingolipídeo/genética , Transdução de Sinais , Esfingosina/análogos & derivados , Esfingosina/fisiologia , Receptores de Esfingosina-1-Fosfato , Linfócitos T/fisiologia
12.
Proc Natl Acad Sci U S A ; 107(23): 10638-43, 2010 Jun 08.
Artigo em Inglês | MEDLINE | ID: mdl-20479237

RESUMO

Cholera toxin (CT) elicits a mucosal immune response in mice when used as a vaccine adjuvant. The mechanisms by which CT exerts its adjuvant effects are incompletely understood. We show that protection against inhalation anthrax by an irradiated spore vaccine depends on CT-mediated induction of IL-17-producing CD4 Th17 cells. Furthermore, IL-17 is involved in the induction of serum and mucosal antibody responses by CT. Th17 cells induced by CT have a unique cytokine profile compared with those induced by IL-6 and TGF-beta, and their induction by CT requires cAMP-dependent secretion of IL-1beta and beta-calcitonin gene-related peptide by dendritic cells. These findings demonstrate that Th17 cells mediate mucosal adjuvant effects of CT and identify previously unexplored pathways involved in Th17 induction that could be targeted for development of unique mucosal adjuvants.


Assuntos
Adjuvantes Imunológicos/farmacologia , Vacinas contra Antraz/imunologia , Toxina da Cólera/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Animais , Formação de Anticorpos , Toxina da Cólera/farmacologia , Imunidade nas Mucosas , Inalação , Interleucina-17/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Mucosa/imunologia
13.
Environ Sci Pollut Res Int ; 30(12): 33195-33205, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36478548

RESUMO

Waterlogging due to rain-fed floods is considered a natural calamity worldwide that causes lands in the north region of Bangladesh to remain underwater and uncultivable for most crops during the rainy season. This unused wasteland has immense potential for additional earnings for the marginal farmer by converting it to cultivable land through proper utilization by cultivating water chestnuts. Increasing the productivity and efficiency of water chestnut farming in these wastelands would facilitate higher food production for the growing populations. Therefore, this study estimates the farmers' profitability of water chestnut production along with technical efficiency (TE) and land use efficiency (LUE). Primary data from 150 farmers of Natore, Naogaon, and Jamalpur districts were used and stochastic frontier analysis was employed. Results reveal that no farmers had any training in water chestnut farming. Human labor and land use costs incur more than 80% of the total cost and laborers shortage was found due to skin problems working in the water. However, water chestnut farming was profitable in all districts and the average benefit-cost ratio was 1.37. TE results indicate that there was an opportunity to increase the water chestnut production by 20.2% using the same amount of inputs. LUE by the water chestnut farmers was found to be very low; hence, they were using their land inefficiently and this inefficiency was positively affected by farm size and number of family members and negatively affected by age, year of schooling, and income. The study suggests research focusing on improved management of water chestnut without affecting laborers and training to facilitate higher productivity and LUE.


Assuntos
Eleocharis , Humanos , Bangladesh , Agricultura , Fazendas , Verduras
14.
J Glob Antimicrob Resist ; 33: 177-185, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-36944411

RESUMO

OBJECTIVES: Edwardsiella ictaluri is an important pathogen in farmed raised catfish. Recently, we showed that resistance to tetracycline and florfenicol in the E. ictaluri MS-17-156 strain isolated from channel catfish was facilitated by acquisition of a 135 kb plasmid (named pEIMS-171561). METHODS: We described the genetic structure of pEIMS-171561. Plasmid copy number and stability within E. ictaluri strain MS-17-156 was determined. We also investigated the in vitro and in vivo transferability of pEIMS-171561 using catfish as a model for in vivo transfer. RESULTS: pEIMS-171561 belonged to the IncA/C group and contained florfenicol efflux major facilitator superfamily (MFS) (floR), sulfonamides (sul2), and tetracycline efflux MFS (tetD) genes. The plasmid contained two conjugative transfer-associated regions and encoded six transposases and insertion sequences. In vitro conjugation experiments demonstrated that the IncA/C plasmid can transfer from E. ictaluri to Escherichia coli. The plasmid was stable in E. ictaluri without selection pressure for 33 days. We showed that pEIMS-171561 did not transfer from E. ictaluri MS-17-156 to endogenous microbiota in catfish. Moreover, we could not detect in vivo conjugal transfer of pEIMS-171561 from E. ictaluri to E. coli. Results from real-time PCR revealed upregulation of the floR gene in the intestines of catfish receiving florfenicol-medicated feed, compared with that in catfish receiving unmedicated feed. CONCLUSION: This study demonstrated that pEIMS-171561 did not disseminate from E. ictaluri to gut microbiota under selective pressure. This result suggests a limited role of the fish microbiota as a reservoir for this plasmid and for the spread of resistance.


Assuntos
Peixes-Gato , Infecções por Enterobacteriaceae , Animais , Edwardsiella ictaluri/genética , Escherichia coli/genética , Infecções por Enterobacteriaceae/veterinária , Infecções por Enterobacteriaceae/tratamento farmacológico , Plasmídeos/genética , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Tetraciclina/uso terapêutico , Peixes-Gato/genética , Resistência Microbiana a Medicamentos
15.
Sci Immunol ; 8(82): eabn0484, 2023 04 28.
Artigo em Inglês | MEDLINE | ID: mdl-37115913

RESUMO

The networks of transcription factors (TFs) that control intestinal-resident memory CD8+ T (TRM) cells, including multipotency and effector programs, are poorly understood. In this work, we investigated the role of the TF Bcl11b in TRM cells during infection with Listeria monocytogenes using mice with post-activation, conditional deletion of Bcl11b in CD8+ T cells. Conditional deletion of Bcl11b resulted in increased numbers of intestinal TRM cells and their precursors as well as decreased splenic effector and circulating memory cells and precursors. Loss of circulating memory cells was in part due to increased intestinal homing of Bcl11b-/- circulating precursors, with no major alterations in their programs. Bcl11b-/- TRM cells had altered transcriptional programs, with diminished expression of multipotent/multifunctional (MP/MF) program genes, including Tcf7, and up-regulation of the effector program genes, including Prdm1. Bcl11b also limits the expression of Ahr, another TF with a role in intestinal CD8+ TRM cell differentiation. Deregulation of TRM programs translated into a poor recall response despite TRM cell accumulation in the intestine. Reduced expression of MP/MF program genes in Bcl11b-/- TRM cells was linked to decreased chromatin accessibility and a reduction in activating histone marks at these loci. In contrast, the effector program genes displayed increased activating epigenetic status. These findings demonstrate that Bcl11b is a frontrunner in the tissue residency program of intestinal memory cells upstream of Tcf1 and Blimp1, promoting multipotency and restricting the effector program.


Assuntos
Linfócitos T CD8-Positivos , Fatores de Transcrição , Camundongos , Animais , Linfócitos T CD8-Positivos/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Diferenciação Celular , Intestinos , Proteínas Supressoras de Tumor/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo
16.
Heliyon ; 8(9): e10618, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36158096

RESUMO

The present study was mainly an impact of farm and off-farm activities on household income and participation of rural women in the Thakurgaon district of Bangladesh. The specific aim was to compare the rural women's participation considering their socioeconomic characteristics, income contribution to household income, and its influencing factors. An Independent Sample T-test was used to compare socioeconomic differences. Pearson's correlation test was used to determine the relationship between women's personal income and household income. The propensity Score Matching (PSM) model was used for impact evaluation of off-farm activities. The result of the t-test showed that off-farm activities were significantly ahead for women from the farm women in terms of socioeconomic variables. There was positive and a strong correlation between women's personal income from off-farm activities and their household income. The result of PSM shows that off-farm activities have a positive and significant impact on rural women's income. Estimation of the binary Probit model and marginal effects of related explanatory variables revealed that educational status, family size, work experiences, personal income, saving, and training, significantly affected rural women's involvement in off-farm activities. Therefore, to increase the pace of work, the participation of rural women in off-farm activities needs to be made more effective and efficient, for which government and non-government organizations need to take necessary steps in area-based development (such as work environment, credit facilities, communication, infrastructure, etc.).

17.
Matrix Biol ; 107: 59-76, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35176450

RESUMO

Versican is a large chondroitin sulfate/dermatan sulfate proteoglycan that plays a key role in the formation of the provisional matrix. Here, we generated dextran sulfate sodium-induced colitis in knockin-mice, R/R, expressing ADAMTS-resistant versican, and investigated the impact of accumulating versican and its turnover in the inflammatory colon mucosa. Histologically, R/R colon showed decreased levels of tissue destruction and an increased number of myofibroblasts and macrophages. Characterization of inflammatory cells revealed an increase in F4/80+ macrophages in R/R colon, compared with wildtype, without a clear shift between M1 and M2 populations. Intestinal stroma exhibited a higher number of myofibroblasts in R/R, suggesting increased levels of tissue regeneration. Coculture of macrophages and stromal fibroblasts obtained from inflammatory colon showed that wild-type macrophages inhibited myofibroblastic differentiation of R/R fibroblasts but not wild-type. This inhibitory effect was due to an increased level of versikine, a cleaved fragment of versican by ADAMTS proteinases. Taken together, our results demonstrate versikine as the direct regulator that inhibits repair of inflamed tissue.


Assuntos
Colite , Versicanas , Animais , Colite/induzido quimicamente , Colite/genética , Fibroblastos , Camundongos , Versicanas/genética , Cicatrização
18.
PLoS One ; 17(9): e0274005, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36054213

RESUMO

Listeria monocytogenes is a ubiquitous opportunistic foodborne pathogen capable of survival in various adverse environmental conditions. Pathogenesis of L. monocytogenes is tightly controlled by a complex regulatory network of transcriptional regulators that are necessary for survival and adaptations to harsh environmental conditions both inside and outside host cells. Among these regulatory pathways are members of the DeoR-family transcriptional regulators that are known to play a regulatory role in sugar metabolism. In this study, we deciphered the role of FruR, a DeoR family protein, which is a fructose operon transcriptional repressor protein, in L. monocytogenes pathogenesis and growth. Following intravenous (IV) inoculation in mice, a mutant strain with deletion of fruR exhibited a significant reduction in bacterial burden in liver and spleen tissues compared to the parent strain. Further, the ΔfruR strain had a defect in cell-to-cell spread in L2 fibroblast monolayers. Constitutive activation of PrfA, a pleiotropic activator of L. monocytogenes virulence factors, did not restore virulence to the ΔfruR strain, suggesting that the attenuation was not a result of impaired PrfA activation. Transcriptome analysis revealed that FruR functions as a positive regulator for genes encoding enzymes involved in the pentose phosphate pathway (PPP) and as a repressor for genes encoding enzymes in the glycolysis pathway. These results suggested that FruR may function to facilitate NADPH regeneration, which is necessary for full protection from oxidative stress. Interestingly, deletion of fruR increased sensitivity of L. monocytogenes to H2O2, confirming a role for FruR in survival of L. monocytogenes during oxidative stress. Using anti-mouse neutrophil/monocyte monoclonal antibody RB6-8C5 (RB6) in an in vivo infection model, we found that FruR has a specific function in protecting L. monocytogenes from neutrophil/monocyte-mediated killing. Overall, this work clarifies the role of FruR in controlling L. monocytogenes carbon flow between glycolysis and PPP for NADPH homeostasis, which provides a new mechanism allowing metabolic adaptation of L. monocytogenes to oxidative stress.


Assuntos
Listeria monocytogenes , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica , Peróxido de Hidrogênio/metabolismo , Listeria monocytogenes/genética , Listeria monocytogenes/patogenicidade , Camundongos , Fatores de Terminação de Peptídeos/metabolismo , Regulon , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Virulência
19.
Heliyon ; 7(5): e07152, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-34136702

RESUMO

Financial and factor demand analysis of broiler production has been estimated in this paper using a farm survey data of 210 farmers from four major broiler producing areas (i.e., Dhaka, Rajshahi, Mymensingh and Chittagong) of Bangladesh. Findings showed that broiler farming incurred most of its cost from its operating input, mainly feed. Broiler farming was financially profitable, but the performance of Mymensingh division was comparatively low, arising from a high unit cost of production and low unit price selling than the others. The net return was highest in Dhaka division, while Rajshahi division showed the highest ratio in returns on investment. However, in terms of cost (variable) and net return of broiler farming, no significant difference among the study areas was observed. The value of own price elasticity for feed, chick price, and labour price were negative and inelastic, which were -0.00249, -0.05718, and -0.13101, respectively. Besides, a complementary relationship was found between feed and day-old chick and feed and labour while day-old chick and labour were substitutes. The study also revealed that cross price elasticity was highly inelastic, and changes in the prices of inputs did not result in massive changes in the quantity demanded of other inputs for broiler farming.

20.
J Histochem Cytochem ; 68(11): 763-775, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-33131383

RESUMO

Versican is a large chondroitin sulfate/dermatan sulfate proteoglycan belonging to the aggrecan/lectican family. In adults, this proteoglycan serves as a structural macromolecule of the extracellular matrix in the brain and large blood vessels. In contrast, versican is transiently expressed at high levels during development and under pathological conditions when the extracellular matrix dramatically changes, including in the inflammation and repair process. There are many reports showing the upregulation of versican in cancer, which correlates with cancer aggressiveness. Versican has four classical splice variants, and all the variants contain G1 and G3 domains at N- and C-termini, respectively. There are two glycosaminoglycan attachment domains CSα and CSß. The largest V0 variant contains both CSα and CSß, V1 contains CSß, V2 contains CSα, and the shortest G3 variant has neither of them. Versican degradation is initiated by cleavage at a site in the CSß domain by ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) proteinases. The N-terminal fragment containing the G1 domain has been reported to exert various biological functions, although its mechanisms of action have not yet been elucidated. In this review, we describe the role of versican in inflammation and cancer and also address the biological function of versikine.


Assuntos
Matriz Extracelular/metabolismo , Neoplasias/metabolismo , Versicanas/metabolismo , Animais , Humanos , Inflamação/metabolismo
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