To study the role of intestinal flora in the development of bloodstream infections (BSIs). 42 patients and 19 healthy controls (HCs) were screened into the study and their intestinal flora was measured by 16S rRNA gene sequencing.
The bacterial diversity was significantly lower in the BSI group compared with that in the HCs (P < 0.001), and beta diversity was significantly differentiated between the two groups (PERMANOVA, P = 0.001). The four keystone species [Roseburia, Faecalibacterium, Prevotella, and Enterococcus (LDA > 4)] differed significantly between the two groups. Dysbiosis of fecal microbial ecology is a common condition present in patients with BSI. The proliferation of certain pathogens or reduction of SCFA-producing bacteria would cause susceptibility to BSI.
Over the past two decades, as the importance of gut microbiota to human health has become widely known, attempts have been made to treat diseases by correcting dysbiosis of gut microbiota through fecal microbiota transplantation (FMT). Apart from current knowledge of gut microbiota, FMT to treat disease has a long history, from the treatment of food poisoning in the fourth century to the treatment of Clostridioides difficile infections in the twentieth century. In 2013, FMT was recognized as a standard treatment for recurrent C. difficile because it consistently showed high efficacy. Though recurrent C. difficile is the only disease internationally recognized for FMT efficacy, FMT has been tested for other diseases and shown some promising preliminary results. Different FMT methods have been developed using various formulations and administration routes.
Despite advances in FMT, some issues remain to be resolved, such as donor screening, manufacturing protocols, and unknown components in the fecal microbiota. In this review, we discuss the mechanisms, clinical indications, methods, and challenges of current FMT. We also discuss the development of alternative therapies to overcome the challenges of FMT.
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Dengue virus (DENV), from the Flaviviridae family, is the causative agent of dengue fever and poses a significant global health challenge. The virus primarily affects the vascular system and liver; however, a growing body of evidence suggests its involvement in the gastrointestinal (GI) tract, contributing to clinical symptoms such as abdominal pain, vomiting, and diarrhea. However, the mechanisms underlying DENV infection in the digestive system remain largely unexplored. Prior research has detected viral RNA in the GI tissue of infected animals; however, whether the dengue virus can directly infect human enterocytes remains unclear. In this study, we examine the infectivity of human intestinal cell lines to the dengue virus and their subsequent response. We report that the Caco-2 cell line, a model of human enterocytes, is susceptible to infection and capable of producing viruses. Notably, differentiated Caco-2 cells exhibited a lower infection rate yet a higher level of virus production than their undifferentiated counterparts. These findings suggest that human intestinal cells are a viable target for the dengue virus, potentially elucidating the GI symptoms observed in dengue fever and offering a new perspective on the pathogenetic mechanisms of the virus.
Ubiquitin is highly conserved in most eukaryotes and involved
in diverse physiological processes, including cell division, protein
quality control, and protein degradation mediated by the
ubiquitin-proteasome system after heat shock, glucose-starvation,
and oxidative stress. However, the role of the ubiquitin
gene UBI4, which contains five consecutive head-to-tail ubiquitin
repeats, in meiosis has not been investigated. In this
study, we show that the Saccharomyces cerevisiae polyubiquitin
precursor gene, UBI4, is required to promote synaptonemal
complex (SC) formation and suppress excess doublestrand
break formation. Moreover, the proportion of Zip1
polycomplexes, which indicate abnormal SC formation, in
cells with a mutation in UBI4 (i.e., ubi4Δ cells) is higher than
that of wild-type cells, implying that the UBI4 plays an important
role in the early meiotic prophase I. Interestingly, although
ubi4Δ cells rarely form full-length SCs in the pachytene
stage of prophase I, the Zip3 foci are still seen, as in
wild-type cells. Moreover, ubi4Δ cells proficiently form crossover
and noncrossover products with a slight delay compared
to wild-type cells, suggesting that UBI4 is dispensable in SCcoupled
recombination. Our findings demonstrate that UBI4
exhibits dual functions that are associated with both positive
and negative roles in SC formation and recombination during
meiosis.
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The deubiquitinase Usp7 in
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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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Zaire ebolavirus, commonly called Ebola virus (EBOV), is an
RNA virus that causes severe hemorrhagic fever with high
mortality. Viral protein 35 (VP35) is a virulence factor encoded
in the EBOV genome. VP35 inhibits host innate immune
responses and functions as a critical cofactor for viral
RNA replication. EBOV VP35 contains a short conserved
motif that interacts with dynein light chain 8 (LC8), which
serves as a regulatory hub protein by associating with various
LC8-binding proteins. Herein, we present the crystal structure
of human LC8 bound to the peptide comprising residues
67−76 of EBOV VP35. Two VP35 peptides were found to
interact with homodimeric LC8 by extending the central β-
sheets, constituting a 2:2 complex. Structural analysis demonstrated
that the intermolecular binding between LC8 and
VP35 is mainly sustained by a network of hydrogen bonds
and supported by hydrophobic interactions in which Thr73
and Thr75 of VP35 are involved. These findings were verified
by binding measurements using isothermal titration calorimetry.
Biochemical analyses also verified that residues 67−76
of EBOV VP35 constitute a core region for interaction with
LC8. In addition, corresponding motifs from other members
of the genus Ebolavirus commonly bound to LC8 but with
different binding affinities. Particularly, VP35 peptides originating
from pathogenic species interacted with LC8 with
higher affinity than those from noninfectious species, suggesting
that the binding of VP35 to LC8 is associated with
the pathogenicity of the Ebolavirus species.
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Phosphate sugar isomerases, catalyzing the isomerization between
ketopentose/ketohexose phosphate and aldopentose/
aldohexose phosphate, play an important role in microbial
sugar metabolism. They are present in a wide range of microorganisms.
They have attracted increasing research interest
because of their broad substrate specificity and great potential
in the enzymatic production of various rare sugars. Here,
the enzymatic properties of various phosphate sugar isomerases
are reviewed in terms of their substrate specificities and
their applications in the production of valuable rare sugars because
of their functions such as low-calorie sweeteners, bulking
agents, and pharmaceutical precursor. Specifically, we
focused on the industrial applications of D-ribose-5-phosphate
isomerase and D-mannose-6-phosphate isomerase to
produce D-allose and L-ribose, respectively.
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Bovine mastitis is a common disease in the dairy industry
that causes great economic losses. As the primary pathogen
of contagious mastitis, Staphylococcus aureus (S. aureus) can
invade bovine mammary epithelial cells, thus evading immune
defenses and resulting in persistent infection. Recently,
autophagy has been considered an important mechanism for
host cells to clear intracellular pathogens. In the current study,
autophagy caused by S. aureus was detected, and the correlation
between autophagy and intracellular S. aureus survival
was assessed. First, a model of intracellular S. aureus infection
was established. Then, the autophagy of MAC-T cells was
evaluated by confocal microscopy and western blot. Moreover,
the activation of the PI3K-Akt-mTOR and ERK1/2 signaling
pathways was determined by western blot. Finally, the
relationship between intracellular bacteria and autophagy
was analyzed by using autophagy regulators (3-methyladenine
[3-MA], rapamycin [Rapa] and chloroquine [CQ]). The results showed that S. aureus caused obvious induction of
autophagosome formation, transformation of LC3I/II, and
degradation of p62/SQSTM1 in MAC-T cells; furthermore,
the PI3K-Akt-mTOR and ERK1/2 signaling pathways were
activated. The number of intracellular S. aureus increased
significantly with autophagy activation by rapamycin, whereas
the number decreased when the autophagy flux was inhibited
by chloroquine. Therefore, this study indicated that intracellular
S. aureus can induce autophagy and utilize it to survive
in bovine mammary epithelial cells.
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bacterial infections before the golden age of antibiotics,
but the therapeutic potential of phages was largely ignored
after the discovery of penicillin. Recently, with antibioticresistant
infections on the rise, these phages are receiving renewed
attention to combat problematic bacterial infections.
Our approach is to enhance bacteriophages with antimicrobial
peptides, short peptides with broad-spectrum antibiotic or
antibiofilm effects. We inserted coding sequences for 1018,
an antimicrobial peptide previously shown to be an effective
broad-spectrum antimicrobial and antibiofilm agent, or the
fluorescent marker mCherry, into the T7Select phage genome.
Transcription and production of 1018 or mCherry began
rapidly after E. coli cultures were infected with genetically modified
phages. mCherry fluorescence, which requires a 90 min
initial maturation period, was observed in infected cultures
after 2 h of infection. Finally, we tested phages expressing 1018
(1018 T7) against bacterial planktonic cultures and biofilms,
and found the 1018 T7 phage was more effective than the
unmodified T7Select phage at both killing planktonic cells and
eradicating established biofilms, validating our phage-driven
antimicrobial peptide expression system. The combination
of narrow-spectrum phages delivering relatively high local
doses of broad-spectrum antimicrobials could be a powerful method to combat resistant infections. The experiments we
describe prove this combination is feasible in vitro, but further
testing and optimization are required before genetically modified
phages are ready for use in vivo.
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Zizania latifolia is a perennial herb belonging to the family
Gramineae that has been used as a health food in Asian countries.
In this study, we investigated the antimicrobial effect of
Z. latifolia, which increased human beta-defensin 2 (hBD2)
expression in HaCaT cells. hBD2 expression was further increased
in cells treated with Z. latifolia extracts and subsequently
infected with Staphylococcus aureus. Inversely, S.
aureus infection decreased after treatment. The induction
of hBD2 in HaCaT cells was mediated by the Toll-like receptor
2 (TLR2) signaling pathway, including the activation
of extracellular signal-regulated kinase (ERK) and activator
protein 1 (AP-1). Further study using siRNA revealed that
hBD2 played an important role in the inhibition of S. aureus
infection in HaCaT cells. Our data suggest that Z. latifolia
extracts can be used as an antimicrobial ingredient for skin
treatment formulas.
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Methicillin resistant Staphylococcus aureus (MRSA) with
multiple drug resistance patterns is frequently isolated from
skin and soft tissue infections that are involved in chronic
wounds. Today, difficulties in the treatment of MRSA associated
infections have led to the development of alternative
approaches such as antimicrobial photodynamic therapy. This
study aimed to investigate photoinactivation with cationic
porphyrin derivative compounds against MRSA in in-vitro
conditions. In the study, MRSA clinical isolates with different
antibiotic resistance profiles were used. The newly synthesized
cationic porphyrin derivatives (PM, PE, PPN, and PPL) were used
as photosensitizer, and 655 nm diode laser was used as light
source. Photoinactivation experiments were performed by
optimizing energy doses and photosensitizer concentrations.
In photoinactivation experiments with different energy densities
and photosensitizer concentrations, more than 99% reduction
was achieved in bacterial cell viability. No decrease
in bacterial survival was observed in control groups. It was
determined that there was an increase in photoinactivation
efficiency by increasing the energy dose. At the energy dose
of 150 J/cm2 a survival reduction of over 6.33 log10 was observed
in each photosensitizer type. While 200 μM PM concentration
was required for this photoinactivation, 12.50 μM
was sufficient for PE, PPN, and PPL. In our study, antimicrobial
photodynamic therapy performed with cationic porphyrin
derivatives was found to have potent antimicrobial efficacy
against multidrug resistant S. aureus which is frequently
isolated from wound infections.
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J. Microbiol. 2018;56(9):665-672. Published online August 23, 2018
Nine Klebsiella pneumoniae isolates coproducing NDM-1
and OXA-232 carbapenemases were successively isolated
from a single patient. Although they were isolated simultaneously
and were isogenic, they presented different colony
phenotypes (matt and mucoid). All nine isolates were resistant
to most antibiotics except colistin and fosfomycin. In
addition, matt-type isolates were resistant to tigecycline. No
differences were detected in the cps cluster sequences, except
for the insertion of IS5 in the wzb gene of two matt-type isolates.
In vitro virulence assays based on production of capsular
polysaccharide, biofilm formation, and resistance to
human serum indicated that the mucoid-type isolates were
significantly more virulent than the matt-type. In addition,
mucoid-type isolates showed higher survival rates than the
matt-type ones in infection experiments in the fruit fly, suggesting
a higher virulence of K. pneumoniae isolates with a
mucoid phenotype. To our knowledge, this is the first report
of K. pneumoniae colonies with different phenotypes being
isolated from the same sample. In addition, we show that virulence
varies with colony phenotype. Dissemination of K.
pneumoniae isolates expressing both antibiotic resistance
and high virulence would constitute a great threat.
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the last two decades, which is a historic milestone to understand
the molecular basis of bacterial pathogenesis. They also
provide small-scale research platforms for identification of
virulence factors, screening for antibacterial hits, and evaluation
of antibacterial efficacy. The fruit fly, Drosophila melanogaster
is one of the model hosts for a variety of bacterial
pathogens, in that the innate immunity pathways and tissue
physiology are highly similar to those in mammals. We here
present a relatively simple protocol to assess the key aspects
of the polymicrobial interaction in vivo between the human
opportunistic pathogens, Pseudomonas aeruginosa and Staphylococcus
aureus, which is based on the systemic infection
by needle pricking at the dorsal thorax of the flies. After infection,
fly survival and bacteremia over time for both P.
aeruginosa and S. aureus within the infected flies can be monitored
as a measure of polymicrobial virulence potential.
The infection takes ~24 h including bacterial cultivation. Fly
survival and bacteremia are assessed using the infected flies
that are monitored up to ~60 h post-infection. These methods
can be used to identify presumable as well as unexpected phenotypes
during polymicrobial interaction between P. aeruginosa
and S. aureus mutants, regarding bacterial pathogenesis
and host immunity.
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Pseudomonas aeruginosa has been identified as an important
causative agent of airway infection, mainly in cystic fibrosis.
This disease is characterized by defective mucociliary clearance
induced in part by mucus hyper-production. Mucin is
a major component of airway mucus and is heavily O-glycosylated,
with a protein backbone. Airway infection is known
to be established with bacterial adhesion to mucin. However,
the genes involved in mucin degradation or utilization remain
elusive. In this study, we sought to provide a genetic basis of
P. aeruginosa airway growth by identifying those genes. First,
using RNASeq analyses, we compared genome-wide expression
profiles of PAO1, a prototype P. aeruginosa laboratory
strain, grown in M9-mucin (M9M) and M9-glucose (M9G)
media. Additionally, a PAO1 transposon (Tn) insertion mutants
library was screened for mutants defective in growth
in M9M medium. One mutant with a Tn insertion in the
xcpU gene (PA3100) was determined to exhibit faulty growth
in M9M medium. This gene contributes to the type II secretion
system, suggesting that P. aeruginosa uses this secretion
system to produce a number of proteins to break down and
assimilate the mucin molecule. Furthermore, we screened
the PAO1 genome for genes with protease activity. Of 13 mutants,
one with mutation in PA3247 gene exhibited defective
growth in M9M, suggesting that the PA3247-encoded protease
plays a role in mucin utilization. Further mechanistic
dissection of this particular process will reveal new drug targets,
the inhibition of which could control recalcitrant P. aeruginosa
infections.
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