The envE gene of Salmonella enterica serovar Typhimurium is encoded within Salmonella Pathogenicity Island-11 (SPI-11) and is located immediately upstream of the virulence gene msgA (macrophage survival gene A) in the same transcriptional orientation. To date, the characteristics and roles of envE remain largely unexplored. In this study, we show that EnvE, a predicted lipoprotein, is localized on the outer membrane using sucrose gradient ultracentrifugation. Under oxidative stress conditions, envE transcription is suppressed, while msgA transcription is induced, indicating an inverse correlation between the mRNA levels of the two neighboring genes. Importantly, inactivation of envE leads to constitutive transcription of msgA regardless of the presence of oxidative stress. Moreover, trans-complementation of the envE mutant with a plasmid-borne envE fails to prevent the induction of msgA transcription, suggesting that envE functions as a cis-regulatory element rather than a trans-acting factor. We further show that both inactivation and complementation of envE confer wild-type levels of resistance to oxidative stress by ensuring the expression of msgA. Our data suggest that the S. enterica envE gene encodes an outer membrane lipoprotein, and its transcription represses msgA expression in a cis-acting manner, probably by transcriptional interference, although the exact molecular details are yet unclear.
Lipopolysaccharide (LPS) is a critical component of the extracellular leaflet within the bacterial outer membrane, forming an effective physical barrier against environmental threats in Gram-negative bacteria. After LPS is synthesized and matured in the bacterial cytoplasm and the inner membrane (IM), LPS is inserted into the outer membrane (OM) through the ATP-driven LPS transport (Lpt) pathway, which is an energy-intensive process. A trans-envelope complex that contains seven Lpt proteins (LptA-LptG) is crucial for extracting LPS from the IM and transporting it across the periplasm to the OM. The last step in LPS transport involves the mediation of the LptDE complex, facilitating the insertion of LPS into the outer leaflet of the OM. As the Lpt system plays an essential role in maintaining the impermeability of the OM via LPS decoration, the interactions between these interconnected subunits, which are meticulously regulated, may be potential targets for the development of new antibiotics to combat multidrug-resistant Gram-negative bacteria. In this review, we aimed to provide an overview of current research concerning the structural interactions within the Lpt system and their implications to clarify the function and regulation of LPS transport in the overall process of OM biogenesis.
Additionally, we explored studies on the development of therapeutic inhibitors of LPS transport, the factors that limit success, and future prospects.
Echoviruses belong to the genus Enterovirus in the Picornaviridae family, forming a large group of Enterovirus B (EVB)
within the Enteroviruses. Previously, Echoviruses were classified based on the coding sequence of VP1. In this study,
we performed a reliable phylogenetic classification of 277 sequences isolated from 1992 to 2019 based on the full-length
genomes of Echovirus. In this report, phylogenetic, phylogeographic, recombination, and amino acid variability landscape
analyses were performed to reveal the evolutional characteristics of Echovirus worldwide. Echoviruses were clustered into
nine major clades, e.g., G1–G9. Phylogeographic analysis showed that branches G2–G9 were linked to common strains,
while the branch G1 was only linked to G5. In contrast, strains E12, E14, and E16 clustered separately from their G3 and
G7 clades respectively, and became a separate branch. In addition, we identified a total of 93 recombination events, where
most of the events occurred within the VP1-VP4 coding regions. Analysis of amino acid variation showed high variability in
the a positions of VP2, VP1, and VP3. This study updates the phylogenetic and phylogeographic information of Echovirus
and indicates that extensive recombination and significant amino acid variation in the capsid proteins drove the emergence
of new strains.
Silver nanoparticles (AgNPs) exhibit strong antibacterial activity and do not easily induce drug resistance; however, the
poor stability and biocompatibility in solution limit their widespread application. In this study, AgNPs were modified with
Polygonatum sibiricum Polysaccharide (PSP) to synthesize PSP@AgNPs with good stability, biocompatibility, and antibacterial
activity. When PSP@AgNP synthesis was performed under a reaction time of 70 min, a reaction temperature of 35 °C,
and an AgNO3-
to-PSP volume ratio of 1:1, the synthesized PSP@AgNPs were more regular and uniform than AgNPs, and
their particle size was around 10 nm. PSP@AgNPs exhibited lower cytotoxicity and hemolysis, and stronger bacteriostatic
activity. PSP@AgNPs damage the integrity and internal structure of cells, resulting in the leakage of intracellular nucleic
acids and proteins. The rate of cell membrane damage in Escherichia coli and Staphylococcus aureus treated with PSP@
AgNPs increased by 38.52% and 43.75%, respectively, compared with that of AgNPs. PSP@AgNPs inhibit the activities
of key enzymes related to antioxidant, energy and substance metabolism in cells. The inhibitory effects on the activities of
superoxide dismutase (SOD), catalase (CAT), adenosine triphosphate enzyme (ATPase), malate dehydrogenase (MDH),
and succinate dehydrogenase (SDH) in E. coli and S. aureus cells were significantly higher than those of AgNPs. In addition,
compared with AgNPs, PSP@AgNPs promote faster healing of infected wounds. Therefore, PSP@AgNPs represent
potential antibacterial agents against wound infections.
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The potential role of the gut microbiota in the pathogenesis
of feeding intolerance (FI) remains unclear. Understanding
the role of the gut microbiota could provide a new avenue for
microbiota-targeted therapeutics. This study aimed to explore
the associations between aberrant gut microbiota and FI in
very low or extremely low birth weight (VLBW/ELBW) preterm
infants. In this observational case-control study, VLBW/
ELBW infants were divided into two groups: FI group and
feeding tolerance (FT) group. 16S rRNA gene sequencing was
performed to analyze the gut microbial diversity and composition
of the infants. The differences in the gut microbiota of
the two groups were compared. In total, 165 stool samples
were obtained from 44 infants, among which, 31 developed
FI and 13 served as controls. Alpha diversity was the highest
in the meconium samples of the two groups. LEfSe analysis
revealed that the abundances of Peptostreptococcaceae, Clostridiales
and Clostridia in the FT group were significantly higher
than in the FI group. At the phylum level, the FI group was dominated
by Proteobacteria, and the FT group was dominated
by Firmicutes. The meconium samples of the FI group had
higher proportions of γ-proteobacteria and Escherichia-Shigella
and a lower proportion of Bacteroides compared with the FT
group. Kyoto Encyclopedia of Genes and Genomes (KEGG)
analysis demonstrated that aberrant gut bacteria in the FI group
were strongly associated with dysregulation of C5-Brancheddibasic-
acid-metabolism, protein kinases, and sporulation.
These findings reveal candidate microbial markers to prevent
FI. Increased relative abundances of γ-proteobacteria
and Escherichia-Shigella and decreased abundance of Bacteroides
in meconium were associated with an increased risk
of FI, while Peptostreptococcaceae, Clostridiales and Clostridia
reduced the risk of FI in VLBW/ELBW infants.
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The differences in methanogen abundance and community
composition were investigated between nearshore and offshore
sediments in the South Yellow Sea (SYS). Shannon,
Simpson, and Chao1 indices revealed a higher diversity of
methanogens in the nearshore sediments than in the offshore
sediments. The Mann–Whitney U test demonstrated that the
relative abundance of Methanococcoides was significantly
higher in the offshore sediments, while the relative abundances
of Methanogenium, Methanosarcina, Methanosaeta,
Methanolinea, and Methanomassiliicoccus were significantly
higher in the nearshore sediments (P < 0.05). The abundance
of the mcrA gene in the nearshore sediments was significantly
higher than that in the offshore sediments. Furthermore, a
similar vertical distribution of the methanogen and sulfatereducing
bacteria (SRB) abundances was observed in the SYS
sediments, implying there is potential cooperation between
these two functional microbes in this environment. Finally,
total organic carbon (TOC) was significantly correlated with
methanogen community composition.
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Protein phosphatase (PPase) inhibition assay (PPIA) is widely
used to analyze the concentration of microcystins (MCs) because
it is comparatively less expensive and faster than other
assays. This study aimed to optimize the PPIA by determining
a suitable reaction terminator and an optimal methanol
concentration in the sample. The most suitable reaction time
was 90 min, with the corresponding methanol concentration
in the sample being 15% or less. When p-nitrophenyl phosphate
(pNPP) was used as a substrate, copper chloride solution
was suitably used as a reaction terminator, and when 4-
methylumbelliferyl phosphate (MUP) was used, a glycine buffer
not only increased the measurement sensitivity of the reaction
product but also terminated the enzymatic reaction.
When PPase 1 and MUP were used as an enzyme and a substrate,
respectively, the limit of quantitation for MC-leucine/
arginine (LR) was 0.02 μg/L, whereas it was 0.1 μg/L when
pNPP was used as a substrate. The proposed method facilitated
the measurement of MC-LR concentration without
additional pretreatments, such as concentration or purification;
therefore, this method was suitable and feasible for the
continuous monitoring of MCs in drinking water.
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Tuberculosis, an infectious disease, is caused by Mycobacterium
tuberculosis. It remains a significant public health issue
around the globe, causing about 1.8 million deaths every year.
Drug-resistant M. tuberculosis, including multi-drug-resistant
(MDR), extremely-drug-resistant (XDR), and totally drugresistant
(TDR) M. tuberculosis, continues to be a threat to
public health. In the case of antibiotic-resistant tuberculosis,
the treatment effect of conventional antibiotics is low. Side
effects caused by high doses over a long period are causing
severe problems. To overcome these problems, there is an urgent
need to develop a new anti-tuberculosis drug that is different
from the existing compound-based antibiotics. Probiotics
are defined as live microorganisms conferring health
benefits. They can be potential therapeutic agents in this context
as the effectiveness of probiotics against different infectious
diseases has been well established. Here, we report that
Lactobacillus crispatus PMC201 shows a promising effect on
tuberculosis isolated from vaginal fluids of healthy Korean
women. Lactobacillus crispatus PMC201 reduced M. tuberculosis
H37Rv under co-culture conditions in broth and reduced
M. tuberculosis H37Rv and XDR M. tuberculosis in macrophages.
Lactobacillus crispatus PMC201 was not toxic to a
guinea pig model and did not induce dysbiosis in a human
intestinal microbial ecosystem simulator. Taken together, these results indicate that L. crispatus PMC201 can be a promising
alternative drug candidate in the current tuberculosis drug
regime. Further study is warranted to assess the in vivo efficacy
and confirm the mode of action of L. crispatus PMC201.
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The VosA-VelB heterocomplex governs expression of several
genes associated with fungal development and secondary
metabolism. In this study, we have investigated the functions
of one of the VosA-VelB-activated developmental genes vadJ
in development and production of the mycotoxin sterigmatocystin
in the model fungus Aspergillus nidulans. The vadJ
gene is predicted to encode a 957-amino acid length protein
containing a highly conserved sensor histidine kinase domain.
The deletion of vosA or velB resulted in decreased mRNA
levels of vadJ throughout the life cycle, suggesting that VosA
and VelB are necessary for proper expression of vadJ. Nullifying
vadJ led to highly restricted colony growth, lowered formation
of asexual spores, and about two-fold reduction in
conidial viability. Conversely, the deletion of vadJ resulted in
elevated production of sexual fruiting bodies and sterigmatocystin.
These suggest that VadJ is necessary for proper coordination
of asexual and sexual development, and sterigmatocystin
production. In accordance with this idea, the deletion
of vadJ led to elevated mRNA levels of the two key sexual
developmental activators esdC and nsdD. In summary, the
putative sensor histidine kinase VadJ represses sexual development
and sterigmatocystin production, but activates
asexual development in A. nidulans.
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Dragon bamboo (Dendrocalamus giganteus) is a giant sympodial
bamboo species widely distributed in Asia. However,
it remains unclear how dragon bamboo and soil microbes interact
to affect soil properties. In this study, we investigated
the planting patterns (semi-natural and artificial) on different
slopes (sunny and shady) to determine the effects on soil properties
and microbial community. The results showed that
the soil in which dragon bamboo was grown was acidic, with
a pH value of ~5. Also, the soil organic matter content, nitrogen
hydrolysate concentration, total nitrogen, available potassium,
and total potassium of the dragon bamboo seminatural
forest significantly improved, especially on the sunny
slope. In contrast, the available phosphorus level was higher
in the artificial bamboo forest, probably owing to the phosphate
fertilizer application. The bacterial and fungal diversity
and the bacterial abundance were all higher on the sunny
slope of the semi-natural forest than those in the other samples.
The microbial operational taxonomic units (OTUs)
shared between the shady and sunny slopes accounted for
47.8–62.2%, but the core OTUs of all samples were only 24.4–
30.4% of each sample, suggesting that the slope type had a
significant effect on the microbial community. Some acidophilic
microbes, such as Acidobacteria groups, Streptomyces
and Mortierella, became dominant in dragon bamboo forest
soil. A PICRUSt analysis of the bacterial functional groups
revealed that post-translational modification, cell division,
and coenzyme transport and metabolism were abundant in
the semi-natural forest. However, some microorganisms with
strong stress resistance might be activated in the artificial
forest. Taken together, these results illustrated the influence
of dragon bamboo growth on soil physicochemical property
and microbial community, which might help understand the
growth status of dragon bamboo under different planting
patterns.
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The soil organic carbon (SOC) mineralization rate in sandy
soil plays an important role in improving soil quality, and a
research is needed to determine management practices that
optimize the mineralization rate. When sandy soil is improved
by adding soft rock, the specific promotion process of bacterium
to SOC mineralization remain unclear. To investigate
these mechanisms, we selected four treatments with soft
rock to sand volume ratios of 0:1 (CK), 1:5 (C1), 1:2 (C2)
and 1:1 (C3) to study. The mineralization rate of organic carbon
was measured using the lye absorption method. Highthroughput
sequencing and scanning electron microscopy
were used to determine the bacterial community structure
and soil microstructure, respectively. The results showed that
the organic carbon content of the sandy soil increased significantly
(182.22–276.43%) after using the soft rock treatments.
The SOC mineralization rate could be divided into two
stages: a rapid decline during days 1–8 and a slow decline
during days 8–60. With increased incubation time, the intensity
of the cumulative release of organic carbon gradually
weakened. Compared with the CK treatment, the SOC mineralization
accumulation (Ct) and the potential mineralizable
organic carbon content (C0) in the C1, C2, and C3 treatments
increased significantly, by 106.98–225.94% and 112.22–
254.08%, respectively. The cumulative mineralization rate (Cr)
was 18.11% and 21.38% smaller with treatments C2 and C3,
respectively. The SOC mineralization rate constant (k) decreased
significantly after the addition of soft rock, while the
half-turnover period (Th) changed inversely with k. Compared
with the CK treatment, the number of gene copies of
the soil bacteria increased by 15.38–272.53% after adding soft
rock, with the most significant increase in treatment C3. The
bacterial diversity index also increased significantly under
treatment C3. The three dominant bacteria were Proteobacteria,
Actinobacteria, and Chloroflexi. The correlation between
Cr and one of the non-dominant bacteria, Firmicutes,
was large, and the bacteria had a significant positive correlation
with k. At the same time, the abundance of Firmicutes
under treatments C2 and C3 was small. As the proportion
of soft rock increased, the soil particles changed from point
contact to surface contact, and the adhesion on the surface
of the particles gradually increased. Results from this study
show that the retention time of SOC can be increased and
the carbon sequestration effect is better when the ratio of
soft rock to sand is set to 1:2.
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Outer membrane vesicles (OMVs) are nanostructures of 20–
200 nm diameter deriving from the surface of several Gramnegative
bacteria. OMVs are emerging as shuttles involved in
several mechanisms of communication and environmental
adaptation. In this work, OMVs were isolated and characterized
from Novosphingobium sp. PP1Y, a Gram-negative
non-pathogenic microorganism lacking LPS on the outer
membrane surface and whose genome was sequenced and
annotated. Scanning electron microscopy performed on samples
obtained from a culture in minimal medium highlighted
the presence of PP1Y cells embedded in an extracellular matrix
rich in vesicular structures. OMVs were collected from
the exhausted growth medium during the mid-exponential
phase, and purified by ultracentrifugation on a sucrose gradient.
Atomic force microscopy, dynamic light scattering and
nanoparticle tracking analysis showed that purified PP1Y
OMVs had a spherical morphology with a diameter of ca. 150
nm and were homogenous in size and shape. Moreover, proteomic
and fatty acid analysis of purified OMVs revealed a
specific biochemical “fingerprint”, suggesting interesting details
concerning their biogenesis and physiological role. Moreover,
these extracellular nanostructures do not appear to be
cytotoxic on HaCaT cell line, thus paving the way to their
future use as novel drug delivery systems.
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were assessed. The results showed that cordycepin effectively
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subtilis, respectively. Scanning electron microscope and
transmission electron microscope examination confirmed
that cordycepin caused obvious damages in the cytoplasmatic
membranes of both E. coli and B. subtilis. Outer membrane
permeability assessment indicated the loss of barrier function
and the leakage of cytoplasmic contents. Propidium
iodide and carboxyfluorescein diacetate double staining approach
coupled with flow cytometry analysis indicated that
the integrity of cell membrane was severely damaged during
a short time, while the intracellular enzyme system still
remained active. This clearly suggested that membrane damage
was one of the reasons for cordycepin efficacy against
bacteria. Additionally, results from circular dichroism and
fluorescence analysis indicated cordycepin could insert to
genome DNA base and double strand, which disordered the
structure of genomic DNA. Basis on these results, the mode
of bactericidal action of cordycepin against E. coli and B.
subtilis was found to be a dual mechanism, disrupting bacterial
cell membranes and binding to bacterial genomic DNA
to interfere in cellular functions, ultimately leading to cell
death.
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The increasing antibiotic resistance of Acinetobacter species
in both natural and hospital environments has become a serious
problem worldwide in recent decades. Because of both
intrinsic and acquired antimicrobial resistance (AMR) against
last-resort antibiotics such as carbapenems, novel therapeutics
are urgently required to treat Acinetobacter-associated infectious
diseases. Among the many pathogenic Acinetobacter
species, A. baumannii has been reported to be resistant to all
classes of antibiotics and contains many AMR genes, such as
blaADC (Acinetobacter-derived cephalosporinase). The AMR
of pathogenic Acinetobacter species is the result of several
different mechanisms, including active efflux pumps, mutations
in antibiotic targets, antibiotic modification, and low
antibiotic membrane permeability. To overcome the limitations
of existing drugs, combination theraphy that can increase
the activity of antibiotics should be considered in the
treatment of Acinetobacter infections. Understanding the
molecular mechanisms behind Acinetobacter AMR resistance
will provide vital information for drug development and
therapeutic strategies using combination treatment. Here,
we summarize the classic mechanisms of Acinetobacter AMR,
along with newly-discovered genetic AMR factors and currently
available antimicrobial adjuvants that can enhance drug
efficacy in the treatment of A. baumannii infections.
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and benzo[a]pyrene. In this study, using transmission electron
microscopy, we confirmed that N. pentaromativorans
US6-1 produces outer membrane vesicles (OMVs). N. pentaromativorans
OMVs (hereafter OMVNovo) are spherical in
shape, and the average diameter of OMVNovo is 25–70 nm.
Proteomic analysis revealed that outer membrane proteins
and periplasmic proteins of N. pentaromativorans are the
major protein components of OMVNovo. Comparative proteomic
analysis with the membrane-associated protein fraction
and correlation analysis demonstrated that the outer
membrane proteins of OMVNovo originated from the membrane-
associated protein fraction. To the best of our knowledge,
this study is the first to characterize OMV purified
from halophilic marine bacteria.
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