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1.
Front Microbiol ; 15: 1301204, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38591032

RESUMO

Introduction: Mycobacterium tuberculosis (Mtb), the main cause of tuberculosis (TB), has brought a great burden to the world's public health. With the widespread use of Mtb drug-resistant strains, the pressure on anti-TB treatment is increasing. Anti-TB drugs with novel structures and targets are urgently needed. Previous studies have revealed a series of CYPs with important roles in the survival and metabolism of Mtb. However, there is little research on the structure and function of CYP138. Methods: In our study, to discover the function and targetability of CYP138, a cyp138-knockout strain was built, and the function of CYP138 was speculated by the comparison between cyp138-knockout and wild-type strains through growth curves, growth status under different carbon sources, infection curves, SEM, MIC tests, quantitative proteomics, and lipidomics. Results and discussion: The knockout of cyp138 was proven to affect the Mtb's macrophage infection, antibiotics susceptibility, and the levels of fatty acid metabolism, membrane-related proteins, and lipids such as triacylglycerol. We proposed that CYP138 plays an important role in the synthesis and decomposition of lipids related to the cell membrane structure as a new potential anti-tuberculosis drug target.

2.
Food Microbiol ; 120: 104483, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38431328

RESUMO

Wooden vats are used in the production of some traditional cheeses as the biofilms on wooden vat surfaces are known to transfer large quantities of microbes to cheese. However, the safety of using wooden vats for cheese production remains controversial as the porous structure of wood provides an irregular surface that may protect any attached pathogen cells from cleaning and sanitation processes. On the other hand, the absence of pathogens in wooden vats has been reported in multiple studies and wooden materials have not been associated with foodborne illness outbreaks. The present study determined the survival of Listeria monocytogenes and Shiga toxin-producing Escherichia coli (STEC) during the production of an uncooked pressed cheese in wooden vats as well as their ability to transfer to the wood and then to milk used in subsequent batches of cheese production in the absence of formal cleaning. Results from the study indicate that pathogens inoculated in milk grew during production of the uncooked cheese, but showed limited ability to colonize the wooden vats and contaminate subsequent batches. These results suggest that the risks of using wooden vats to produce cheese is low if the milk is of high microbiological quality.


Assuntos
Queijo , Listeria monocytogenes , Escherichia coli Shiga Toxigênica , Animais , Queijo/microbiologia , Leite/microbiologia , Dinâmica Populacional , Microbiologia de Alimentos
3.
Exp Dermatol ; 32(12): 2102-2111, 2023 12.
Artigo em Inglês | MEDLINE | ID: mdl-37846925

RESUMO

Acne is a chronic disease that often persists for years. Skin microbial communities play an essential role in the development of acne. However, limited information is available about the dynamic patterns of skin microbiota in acne. This study aimed to characterize microbial community changes in skin pores and surfaces of acne patients with varying disease time. In this study, a total of 70 skin samples from 22 subjects were collected and sequenced using 16S rRNA amplicon sequencing. Although microbial compositions in skin pores were similar over time, significant differences in microbial structure were observed on the skin surface, with the dominance of Cutibacterium in the first 3 years and replacement by Staphylococcus in 4-6 years. Lactobacillus and Acinetobacter were more abundant in the normal group and continuingly decreased with disease time on the skin surface. Microbial networks further revealed substantial increases in microbial interactions in the 4-6 years group in both skin surfaces and pores. These results demonstrate that the skin microbiota alters with the disease duration and may provide a potential guide in redirecting skin microbiota towards healthy states.


Assuntos
Acne Vulgar , Microbiota , Humanos , RNA Ribossômico 16S/genética , Estudos Transversais , Acne Vulgar/microbiologia , Pele/microbiologia , Microbiota/genética , Estudos de Coortes
4.
Acta Pharm Sin B ; 13(9): 3678-3693, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37719365

RESUMO

Polymyxin B and polymyxin E (colistin) are presently considered the last line of defense against human infections caused by multidrug-resistant Gram-negative organisms such as carbapenemase-producer Enterobacterales, Acinetobacter baumannii, and Klebsiella pneumoniae. Yet resistance to this last-line drugs is a major public health threat and is rapidly increasing. Polymyxin S2 (S2) is a polymyxin B analogue previously synthesized in our institute with obviously high antibacterial activity and lower toxicity than polymyxin B and colistin. To predict the possible resistant mechanism of S2 for wide clinical application, we experimentally induced bacterial resistant mutants and studied the preliminary resistance mechanisms. Mut-S, a resistant mutant of K. pneumoniae ATCC BAA-2146 (Kpn2146) induced by S2, was analyzed by whole genome sequencing, transcriptomics, mass spectrometry and complementation experiment. Surprisingly, large-scale genomic inversion (LSGI) of approximately 1.1 Mbp in the chromosome caused by IS26 mediated intramolecular transposition was found in Mut-S, which led to mgrB truncation, lipid A modification and hence S2 resistance. The resistance can be complemented by plasmid carrying intact mgrB. The same mechanism was also found in polymyxin B and colistin induced drug-resistant mutants of Kpn2146 (Mut-B and Mut-E, respectively). This is the first report of polymyxin resistance caused by IS26 intramolecular transposition mediated mgrB truncation in chromosome in K. pneumoniae. The findings broaden our scope of knowledge for polymyxin resistance and enriched our understanding of how bacteria can manage to survive in the presence of antibiotics.

5.
Food Microbiol ; 115: 104319, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37567629

RESUMO

Wooden vats are used in the production of some traditional cheeses as the biofilms on wooden vat surfaces are known to transfer large quantities of microbes to cheese. Variability in microbial communities on wooden vats could lead to inconsistent cheese production. In the present study, the influences of environmental conditions and milk type (raw or heat-treated) on the microbial composition of vat biofilms and cheeses made in the vats were studied using amplicon sequencing of bacterial 16S rRNA and fungal internal transcribed spacer genes. Results showed that the microbial composition of biofilms was influenced by environmental conditions but not the milk type used in cheese production. The microbial composition of cheeses can be further affected by bacterial contributions from milk and the selective forces of environmental conditions. Results of this study suggest that controlling environmental conditions could maintain a more consistent microbial composition of biofilms on wooden vats and resulting cheeses. The use of wooden vats coupled with heat-treated milk at one or more stages of cheese production might be a viable approach to produce cheese with high microbial diversities and reduce risks of undesirable microbes related to food safety and quality.


Assuntos
Queijo , Microbiota , Animais , Leite/microbiologia , Queijo/microbiologia , RNA Ribossômico 16S/genética , Bactérias/genética , Microbiota/genética , Microbiologia de Alimentos
6.
Microbiol Spectr ; 11(1): e0369922, 2023 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-36622182

RESUMO

Enterococci can cause various infectious diseases, including urinary tract infection, wound infection, and life-threatening endocarditis and meningitis. The emergence and transmission of vancomycin-resistant enterococci (VRE) have presented a challenge to clinical treatment. There is an urgent need to develop new strategies to fight against this pathogen. This study investigated the antibacterial and anti-biofilm activity of celastrol (CEL), a natural product originating from Tripterygium wilfordii Hook F, against enterococci, and its adjuvant capacity of restoring the susceptibility of VRE to vancomycin in vitro and in vivo. CEL inhibited all enterococcus strains tested, with MICs ranging from 0.5 to 4 µg/mL. More than 50% of biofilm was eliminated by CEL at 16 µg/mL after 24 h of exposure. The combination of CEL and vancomycin showed a synergistic effect against all 23 strains tested in checkerboard assays. The combination of sub-MIC levels of CEL and vancomycin showed a synergistic effect in a time-kill assay and exhibited significant protective efficacy in Galleria mellonella larval infection model compared with either drug used alone. The underlying mechanisms of CEL were explored by conducting biomolecular binding interactions and an enzyme inhibition assay of CEL on bacterial cell-division protein FtsZ. CEL presented strong binding and suppression ability to FtsZ, with Kd and IC50 values of 2.454 µM and 1.04 ± 0.17 µg/mL, respectively. CEL exhibits a significant antibacterial and synergic activity against VRE in vitro and in vivo and has the potential to be a new antibacterial agent or adjuvant to vancomycin as a therapeutic option in combating VRE. IMPORTANCE The emergence and transmission of VRE pose a significant medical and public health challenge. CEL, well-known for a wide range of biological activities, has not previously been investigated for its synergistic effect with vancomycin against VRE. In the present study, CEL exhibited antibacterial activity against enterococci, including VRE strains, and restored the activity of vancomycin against VRE in vitro and in vivo. Hence, CEL has the potential to be a new antibacterial adjuvant to vancomycin and could provide a promising therapeutic option in combating VRE.


Assuntos
Enterococos Resistentes à Vancomicina , Vancomicina , Vancomicina/farmacologia , Antibacterianos/farmacologia , Triterpenos Pentacíclicos/farmacologia , Testes de Sensibilidade Microbiana
7.
Food Microbiol ; 109: 104154, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36309450

RESUMO

Wooden vats are used in the production of some traditional cheeses as the biofilms on wooden vat surfaces are known to transfer large quantities of microbes to cheese. However, very few studies have investigated the microbial composition of biofilms on newly developed wooden vats and how communities assemble and evolve. In the present study, the microbial communities of biofilms were characterized over the activation process on new wooden vats using amplicon sequencing of bacterial 16s rRNA and fungal internal transcribed spacer genes. Results showed that microbes from the whey effectively developed on wooden vats. Lactococcus was highly dominant throughout the vat activation process with substantial increases in the relative abundance of Acetobacter and Lactobacillus at the end of the vat activation (day 7). This was in contrast with fungal communities that stabilized early (day 1) and were dominated by Kluyveromyces. Predicted functions corresponded with the different stages of biofilm formation whereby functions associated with biofilm initiation were enriched on day 1 and those associated with growth and maturation were enriched on days 4 and 7. Microbial succession on wooden vat surfaces is expected to be reproducible based on the early onset and dominance of the deterministic process.


Assuntos
Queijo , Microbiota , Animais , Queijo/microbiologia , RNA Ribossômico 16S/genética , Leite/microbiologia , Microbiota/genética , Biofilmes
8.
Nanoscale ; 15(2): 515-521, 2023 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-36519408

RESUMO

Bacterial infection can cause a series of diseases and play a vital role in medical care. Therefore, early diagnosis of pathogenic bacteria is crucial for effective treatment and the prevention of further infection. However, restricted by the current technology, bacterial detection is usually time-consuming and laborious and the samples need tedious processing even to be tested. Herein, we present a terahertz metasensor based on the coupling of electrical and toroidal dipoles to achieve rapid, non-destructive, label-free identification and highly sensitive quantitative detection of the two most common pathogenic bacteria. The reinforcement of the toroidal dipole significantly boosts the light-matter interactions around the surface of the microstructure, and thus the sensitivity and Q factor of the designed metasensor reach as high as 378 GHz per refractive index unit (RIU) and 21.28, respectively. Combined with the aforementioned advantages, the proposed metasensor successfully identified Escherichia coli and Staphylococcus aureus and quantitatively detected four concentrations with the lowest detectable concentration being ∼104 cfu mL-1 in the experiment. This work naturally enriches the research on THz metasensors based on the interference mechanism and inspires more innovations to facilitate the development of biosensing applications.


Assuntos
Técnicas Biossensoriais , Infecções Estafilocócicas , Humanos , Limite de Detecção , Escherichia coli , Bactérias
9.
Cells ; 11(20)2022 10 13.
Artigo em Inglês | MEDLINE | ID: mdl-36291085

RESUMO

Non-alcoholic fatty liver disease affects one-fourth of the world's population. Central to the disease progression is lipid accumulation in the liver, followed by inflammation, fibrosis and cirrhosis. The underlying mechanism behind the early stages of the disease is poorly understood. We have exposed human hepatic HepG2/C3A cells-based spheroids to 65 µM oleic acid and 45 µM palmitic acid and employed proteomics and lipidomics analysis to investigate their effect on hepatocytes. The treatment successfully induced in vivo hallmarks of NAFLD, as evidenced by intracellular lipid accumulation and increased ATP levels. Quantitative lipidome analysis revealed an increase in ceramides, LPC and saturated triglycerides and a decrease in the ratio of PC/PE, similar to the changes observed in patients' liver biopsies. The proteomics analysis combined with qPCR showed increased epithelial to mesenchymal transition (EMT) signalling. Activation of EMT was further validated by transcriptomics in TGF-ß treated spheroids, where an increase in mesenchymal cell markers (N-cadherin and collagen expression) was found. Our study demonstrates that this model system thus closely echoes several of the clinical features of non-alcoholic fatty liver disease and can be used to investigate the underlying molecular changes occurring in the condition.


Assuntos
Lipidômica , Hepatopatia Gordurosa não Alcoólica , Humanos , Trifosfato de Adenosina/metabolismo , Caderinas/metabolismo , Ceramidas/metabolismo , Transição Epitelial-Mesenquimal , Hepatopatia Gordurosa não Alcoólica/metabolismo , Ácido Oleico/metabolismo , Ácido Palmítico/metabolismo , Proteoma/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Triglicerídeos/metabolismo , Células Hep G2
10.
Mater Horiz ; 9(12): 2984-2992, 2022 11 28.
Artigo em Inglês | MEDLINE | ID: mdl-36073353

RESUMO

Metasurfaces with a strongly enhanced local field are envisioned as a powerful platform for ultrasensitive optical sensors to significantly amplify imperceptible differences between compatible bioanalytes. Through the use of phototunable silicon-based terahertz (THz) metasurfaces, we experimentally demonstrate ultrafast switchable sensing functions. It is found that the THz responses of the coupled-resonances in the metasurfaces shift from Lorentz-lattice mode to electromagnetism-induced transparency (EIT) mode under optical pumping within an ultrashort time of 32 ps, enabling an ultrafast sensitive sensor. For the Lorentz-lattice mode, the THz time-domain signal directly shows a highly sensitive response to detect tiny analytes without extra Fourier transformation as the mismatch between the two modes increases. Once the metasurfaces are switched to the EIT mode, the silicon-metal hybrid structure supports frequency-domain sensing ability due to strong field confinement with a sensitivity of 118.4 GHz/RIU. Both of the sensing configurations contribute to more subtle information and guarantee the accuracy of the sensor performance. Combined with the aforementioned advantages, the proposed metasurfaces have successfully identified colorectal cells between normal, adenoma, and cancer states in experiments. This work furnishes a new paradigm of constructing reliable and flexible metasurface sensors and can be extended to other optics applications.


Assuntos
Técnicas Biossensoriais , Neoplasias Colorretais , Humanos , Neoplasias Colorretais/diagnóstico , Silício
11.
Front Cell Dev Biol ; 10: 979262, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36176276

RESUMO

Background: Lung adenocarcinoma (LUAD) is the most common type of lung cancer with a complex tumor microenvironment. Neddylation, as a type of post-translational modification, plays a vital role in the development of LUAD. To date, no study has explored the potential of neddylation-associated genes for LUAD classification, prognosis prediction, and treatment response evaluation. Methods: Seventy-six neddylation-associated prognostic genes were identified by Univariate Cox analysis. Patients with LUAD were classified into two patterns based on unsupervised consensus clustering analysis. In addition, a 10-gene prognostic signature was constructed using LASSO-Cox and a multivariate stepwise regression approach. Results: Substantial differences were observed between the two patterns of LUAD in terms of prognosis. Compared with neddylation cluster2, neddylation cluster1 exhibited low levels of immune infiltration that promote tumor progression. Additionally, the neddylation-related risk score correlated with clinical parameters and it can be a good predictor of patient outcomes, gene mutation levels, and chemotherapeutic responses. Conclusion: Neddylation patterns can distinguish tumor microenvironment and prognosis in patients with LUAD. Prognostic signatures based on neddylation-associated genes can predict patient outcomes and guide personalized treatment.

12.
J Lipid Res ; 63(9): 100251, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35841948

RESUMO

The outer membrane (OM) of Gram-negative bacteria is an evolving antibiotic barrier composed of a glycerophospholipid (GP) inner leaflet and a lipopolysaccharide (LPS) outer leaflet. The two-component regulatory system CrrAB has only recently been reported to confer high-level polymyxin resistance and virulence in Klebsiella pneumoniae. Mutations in crrB have been shown to lead to the modification of the lipid A moiety of LPS through CrrAB activation. However, functions of CrrAB activation in the regulation of other lipids are unclear. Work here demonstrates that CrrAB activation not only stimulates LPS modification but also regulates synthesis of acyl-glycerophosphoglycerols (acyl-PGs), a lipid species with undefined functions and biosynthesis. Among all possible modulators of acyl-PG identified from proteomic data, we found expression of lipid A palmitoyltransferase (PagP) was significantly upregulated in the crrB mutant. Furthermore, comparative lipidomics showed that most of the increasing acyl-PG activated by CrrAB was decreased after pagP knockout with CRISPR-Cas9. These results suggest that PagP also transfers a palmitate chain from GPs to PGs, generating acyl-PGs. Further investigation revealed that PagP mainly regulates the GP contents within the OM, leading to an increased ratio of acyl-PG to PG species and improving OM hydrophobicity, which may contribute to resistance against certain cationic antimicrobial peptides resistance upon LPS modification. Taken together, this work suggests that CrrAB regulates the palmitoylation of PGs and lipid A within the OM through upregulated PagP, which functions together to form an outer membrane barrier critical for bacterial survival.


Assuntos
Proteínas de Escherichia coli , Lipoilação , Aciltransferases/metabolismo , Antibacterianos , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Glicerofosfatos , Glicerofosfolipídeos , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/metabolismo , Lipídeo A/metabolismo , Lipopolissacarídeos/metabolismo , Lipopolissacarídeos/farmacologia , Palmitatos/metabolismo , Polimixinas/metabolismo , Proteômica
13.
Nanoscale ; 14(27): 9681-9685, 2022 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-35723251

RESUMO

Terahertz (THz) plasmonic resonance based on an arbitrarily designed resonance metasurface is the key technique of choice for enhancing fingerprint absorption spectroscopy identification of biomolecules. Here, we report a broadband THz micro-photonics sensor based on a pixelated frequency-agile metasurface and illustrate its application ability to enhance and differentiate the detection of broadband absorption fingerprint spectra. The design uses symmetrical metal C-shape resonators with the functional graphene micro-ribbons selectively patterned into the gaps. A strong electric resonance with a high quality factor was formed, consisting of an electric dipole mode associated with the excitation of a dark toroidal dipole (TD) mode through the coupling from the electric dipole moment of the individual frequency-agile meta-unit. The resonance positions are nearly linearly modulated with the varying Fermi level of graphene. The configuration arranges a certain metapixel of the metasurface to multiple response spectra assembling a one-to-many mapping between spatial and spectral information which is instrumental in greatly shrinking the actual size of the sensor. By the synchronous regulation of graphene and C-shape rings, we have obtained highly surface-sensitive resonances over a wide spectral range (∼1.5 THz) with a spectral resolution less than 20 GHz. The target multiple enhanced absorption spectrum of glucose molecules is read out in a broadband region with high sensitivity. More importantly, the design can be extended to cover a larger spectral region by altering the range of geometrical parameters. Our microphotonic technique can resolve absorption fingerprints without the need for spectrometry and frequency scanning, thereby providing an approach for highly sensitive and versatile miniaturized THz spectroscopy devices.

14.
iScience ; 25(4): 104091, 2022 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-35378860

RESUMO

The phenotype of ß-thalassemia underlies multigene interactions, making clinical stratification complicated. An increasing number of genetic modifiers affecting the disease severity have been identified, but are still unable to meet the demand of precision diagnosis. Here, we systematically conducted a comparative plasma proteomic profiling on patients with ß-thalassemia and healthy controls. Among 246 dysregulated proteins, 13 core protein signatures with excellent biomarker potential are proposed. The combination of proteome and patients' clinical data revealed patients with codons 41/42 -TTCT mutations have an elevated risk of higher iron burden, dysplasia, and osteoporosis than patients with other genotypes. Notably, 85 proteins correlating to fetal hemoglobin (Hb F) were identified, among which the abundance of 27 proteins may affect the transfusion burden in patients with ß-thalassemia. The current study thus provides protein signatures as potential diagnostic biomarkers or therapeutic clues for ß-thalassemia.

15.
Front Microbiol ; 13: 980217, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36619997

RESUMO

Antivirulence strategy has been developed as a nontraditional therapy which would engender a lower evolutionary pressure toward the development of antimicrobial resistance. However, the majority of the antivirulence agents currently in development could not meet clinical needs due to their narrow antibacterial spectrum and limited indications. Therefore, our main purpose is to develop broad-spectrum antivirulence agents that could target on both Gram-positive and Gram-negative pathogens. We discovered ML364, a novel scaffold compound, could inhibit the productions of both pyocyanin of Pseudomonas aeruginosa and staphyloxanthin of Staphylococcus aureus. Further transcriptome sequencing and enrichment analysis showed that the quorum sensing (QS) system of pathogens was mainly disrupted by ML364 treatment. To date, autoinducer-2 (AI-2) of the QS system is the only non-species-specific signaling molecule that responsible for the cross-talk between Gram-negative and Gram-positive species. And further investigation showed that ML364 treatment could significantly inhibit the sensing of AI-2 or its nonborated form DPD signaling in Vibrio campbellii MM32 and attenuate the biofilm formation across multi-species pathogens including Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae and Staphylococcus aureus. The results of molecular docking and MM/GBSA free energy prediction showed that ML364 might have higher affinity with the receptors of DPD/AI-2, when compared with DPD molecule. Finally, the in vivo study showed that ML364 could significantly improve the survival rates of systemically infected mice and attenuate bacterial loads in the organs of mice. Overall, ML364 might interfere with AI-2 quorum sensing system to exert broad-spectrum antivirulence effect both in vitro and in vivo.

16.
Nanoscale ; 13(44): 18467-18472, 2021 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-34726683

RESUMO

An ultra-sensitive THz metasensor is presented based on quasi-BIC Fano resonance, which can distinguish extremely dilute concentrations (nM) of solutions. It provides a nondestructive sensing approach for disease prevention and diagnosis. However, the main drawback limiting the performance of THz-based bio-chemical sensors is the weak interaction between the optical field and the analyte, the characteristic scale of which is mismatched with the THz wavelength, leading to low sensitivity. Herein, we present an ultra-sensitive THz metasensor based on an electric Fano resonant metasurface which consists of three gold microrods arranged periodically. The designed electric Fano resonance provides a strong near-field enhancement near the surface of the microstructure, significantly boosting the light-analyte interactions and thus the sensitivity. Such an electric Fano resonance is formed by the interference between a leaky electric dipole resonance and a bound toroidal dipole mode which is a symmetry-protected bound state in the continuum supported by the sub-diffractive periodic system here. Owing to the strong electric fields generated near the interface of our microstructure around the toroidal dipole BIC, the proposed structure can distinguish extremely dilute concentrations (nM) of solutions. Importantly, by controlling the degree of geometrical asymmetry, the BIC-inspired mechanism provides an important and simple tool to engineer and tailor the linewidth and Q-factor of our proposed electric Fano resonance, indicating the ability to realize different biosensors for different optical regimes. Our results open new possibilities to realize a non-destructive and non-contact quantitative inspection of low-concentration solutions, providing a useful sensing approach for disease prevention and diagnosis.

17.
mSystems ; 6(5): e0083021, 2021 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-34581601

RESUMO

Prior to the advent of milk pasteurization and the use of defined-strain starter cultures, the production and ripening of cheese relied on the introduction and growth of adventitious microbes from the environment. This study characterized microbial community structures throughout a traditional farmstead cheese production continuum and evaluated the role of the environment in microbial transfer. In total, 118 samples (e.g., raw milk, cheese, and environmental surfaces) were collected from milk harvesting through cheese ripening. Microbial communities were characterized based on amplicon sequencing of bacterial 16S rRNA and fungal internal transcribed spacer genes using the Illumina MiSeq platform. Results indicated that the environment in each processing room harbored unique microbial ecosystems and consistently contributed microbes to milk, curd, and cheese. The diverse microbial composition of milk was initially attributed to milker hands and cow teats and then changed substantially following overnight ripening in a wooden vat to one dominated by lactic acid bacteria, including Lactococcus lactis, Lactobacillus, and Leuconostoc, as well as fungi such as Exophiala, Kluyveromyces, and Candida. Additional microbial contributions were attributed to processing tools, but the composition of the cheese paste remained relatively stable over 60 days of ripening. In contrast, rind communities that were largely influenced by direct contact with bamboo aging mats showed a distinct succession pattern compared to the interior sections. Overall, these findings highlight the critical role of traditional tools and practices in shaping the microbial composition of cheese and broaden our understanding of processing environments as important sources of microbes in food. IMPORTANCE Throughout the 20th century, especially in the United States, sanitation practices, pasteurization of milk, and the use of commercial defined-strain starter cultures have enhanced the safety and consistency of cheese. However, these practices can reduce cheese microbial diversity. The rapid growth of the artisanal cheese industry in the United States has renewed interest in recapturing the diversity of dairy products and the microbes involved in their production. Here, we demonstrate the essential role of the environment, including the use of wooden tools and cheesemaking equipment, as sources of dominant microbes that shape the fermentation and ripening processes of a traditional farmstead cheese produced without the addition of starter cultures or direct inoculation of any other bacteria or fungi. These data enrich our understanding of the microbial interactions between products and the environment and identify taxa that contribute to the microbial diversity of cheese and cheese production.

18.
J Antibiot (Tokyo) ; 74(12): 863-873, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34480092

RESUMO

Antimicrobial resistance has been an increasingly serious threat to global public health. Anti-virulence strategies are being developed to manage antibiotic resistance because they apply a lower selective pressure for antimicrobial-resistant pathogens than that created using traditional bactericides. We aimed to discover novel small molecules that can reduce the production of virulence factors in Pseudomonas aeruginosa and determine the mechanism of action underlying these effects. A clinical compound library was screened, and ostarine was identified as a potential anti-virulence agent. The effects of ostarine were studied via antimicrobial susceptibility testing, bacterial growth assays, pyocyanin quantitation assays, transcriptomic analysis, quorum sensing signal molecule quantification, and real-time PCR assays. Ostarine treatment significantly decreased the synthesis of pyocyanin without any bactericidal action. Besides, ostarine treatment did not affect the relative growth rate and cell morphology of bacteria. Treatment with ostarine interfered with quorum sensing by decreasing the transcription of genes associated with quorum sensing systems and the production of signalling molecules. The inhibition of ostarine on pyocyanin production and gene expression can be alleviated when signalling molecules were supplemented externally. Overall, ostarine may act as a novel anti-virulence agent that can attenuate P. aeruginosa pyocyanin by interfering with quorum sensing systems.


Assuntos
Anilidas/farmacologia , Antibacterianos/farmacologia , Pseudomonas aeruginosa/efeitos dos fármacos , Piocianina/metabolismo , Farmacorresistência Bacteriana , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/patogenicidade , Piocianina/genética , Percepção de Quorum/efeitos dos fármacos , Virulência/efeitos dos fármacos , Fatores de Virulência
19.
Front Cell Infect Microbiol ; 11: 701625, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34395312

RESUMO

Resistance to colistin, especially mobilized colistin resistance (mcr), is a serious threat to public health since it may catalyze a return of the "pre-antibiotic era". Outer membrane vesicles (OMVs) play a role in antibiotic resistance in various ways. Currently, how OMVs participate in mcr-1-mediated colistin resistance has not been established. In this study, we showed that both OMVs from the mcr-1 negative and positive Escherichia coli (E. coli) strains conferred dose-dependent protection from colistin. However, OMVs from the mcr-1 positive strain conferred attenuated protection when compared to the OMVs of a mcr-1 negative strain at the same concentration. The attenuated protective effect of OMVs was related to the reduced ability to absorb colistin from the environment, thus promoting the killing of colistin sensitive E. coli strains. Lipid A modified with phosphoethanolamine was presented in the OMVs of the mcr-1 positive E. coli strain and resulted in decreased affinity to colistin and less protection. Meanwhile, E. coli strain carrying the mcr-1 gene packed more unmodified lipid A in OMVs and kept more phosphoethanolamine modified lipid A in the bacterial cells. Our study provides a first glimpse of the role of OMVs in mcr-1 -mediated colistin resistance.


Assuntos
Colistina , Proteínas de Escherichia coli , Antibacterianos/farmacologia , Colistina/farmacologia , Farmacorresistência Bacteriana , Escherichia coli/genética , Proteínas de Escherichia coli/genética , Testes de Sensibilidade Microbiana , Plasmídeos
20.
Cancer Med ; 10(7): 2509-2522, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33704935

RESUMO

Prostate cancer-associated fibroblasts (prostate CAFs) are essential components of the tumor microenvironment and can promote tumor progression through their immunosuppressive functions. MPSSS, a novel polysaccharide purified from Lentinus edodes, has been reported to have anti-tumor activity. MPSSS could also inhibit the immunosuppressive function of prostate CAFs, which has been demonstrated through that the secretome of MPSSS-treated prostate CAFs could inhibit the proliferation of T cells. However, how the secretome of MPSSS-treated prostate CAFs influence prostate cancer progression is still unclear. Interestingly, we found that the low molecular weight (3-100kD) secretome of prostate CAFs (lmwCAFS) could promote the growth of PC-3 cells, while that of MPSSS-treated prostate CAFs (MT-lmwCAFS) could inhibit their growth. We carried out comparative secretomic analysis of lmwCAFS and MT-lmwCAFS to identify functional molecules that inhibit the growth of PC-3 cells, and proteomic analysis of lmwCAFS-treated PC-3 cells and MT-lmwCAFS-treated PC-3 cells to investigate the underlying molecular mechanism. These analyses suggest that TGF-ß3 from MT-lmwCAFS may inhibit the growth of PC-3 cells. The validated experiments revealed that TGF-ß3 from MT-lmwCAFS activated p21 expression in PC-3 cells by regulating the FoxO pathway thereby inducing G0/G1 cell cycle arrest of PC-3 cells. Overall, our data demonstrated that MPSSS reversed the ability of prostate CAFs to suppress the cell viability of PC-3 cells, which might provide a potential therapeutic strategy to prevent prostate cancer progression.


Assuntos
Fibroblastos Associados a Câncer/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Fatores de Transcrição Forkhead/metabolismo , Polissacarídeos Fúngicos/farmacologia , Neoplasias da Próstata/patologia , Proteômica , Fator de Crescimento Transformador beta3/farmacologia , Actinas/metabolismo , Animais , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/imunologia , Fibroblastos Associados a Câncer/química , Fibroblastos Associados a Câncer/fisiologia , Pontos de Checagem do Ciclo Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Progressão da Doença , Vesículas Extracelulares/química , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Peso Molecular , Células PC-3 , Neoplasias da Próstata/terapia , Cogumelos Shiitake/química , Microambiente Tumoral/imunologia
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