In recent years, the occurrence of antibiotic-resistant pathogens
is increasing rapidly. There is growing concern as
the development of antibiotics is slower than the increase in
the resistance of pathogenic bacteria. Antimicrobial peptides
(AMPs) are promising alternatives to antibiotics. Despite their
name, which implies their antimicrobial activity, AMPs have
recently been rediscovered as compounds having antifungal,
antiviral, anticancer, antioxidant, and insecticidal effects.
Moreover, many AMPs are relatively safe from toxic side effects
and the generation of resistant microorganisms due to
their target specificity and complexity of the mechanisms underlying
their action. In this review, we summarize the history,
classification, and mechanisms of action of AMPs, and
provide descriptions of AMPs undergoing clinical trials. We
also discuss the obstacles associated with the development of
AMPs as therapeutic agents and recent strategies formulated
to circumvent these obstacles.
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Severe acute respiratory syndrome coronavirus 2 (SARSCoV-
2) has caused corona virus disease 2019 (COVID-19)
pandemic and led to mass casualty. Even though much effort
has been put into development of vaccine and treatment methods to combat COVID-19, no safe and efficient
cure has been discovered. Drug repurposing or drug repositioning
which is a process of investigating pre-existing drug
candidates for novel applications outside their original medical
indication can speed up the drug development process.
Raloxifene is a selective estrogen receptor modulator (SERM)
that has been approved by FDA in 1997 for treatment and
prevention of postmenopausal osteoporosis and cancer. Recently,
raloxifene demonstrates efficacy in treating viral infections
by Ebola, influenza A, and hepatitis C viruses and
shows potential for drug repurposing for the treatment of
SARS-CoV-2 infection. This review will provide an overview
of raloxifene’s mechanism of action as a SERM and present
proposed mechanisms of action in treatment of viral infections.
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A polyphasic taxonomic approach was used to characterize
two novel bacterial strains, HDW17AT and HDW17BT, isolated
from the intestine of the diving beetle Cybister lewisianus,
and the dark diving beetle Hydrophilus acuminatus,
respectively. Both strains were Gram-positive and facultative
anaerobic cocci forming cream-colored colonies. The isolates
grew optimally at 25°C, pH 7, in the presence of 0.3% (wt/vol)
NaCl. Phylogenetic analysis based on 16S rRNA gene sequences
and genome sequences showed that the isolates were
members of the genus Vagococcus, and strain HDW17AT was
closely related to Vagococcus fessus CCUG 41755T (98.9% of
16S rRNA gene sequence similarity and 74.3% of average
nucleotide identity [ANI]), whereas strain HDW17BT was
closely related to Vagococcus fluvialis NCFB 2497T (98.9% of
16S rRNA gene sequence similarity and 76.6% of ANI). Both
strains contained C16:0, and C18:1 ω9c as the major cellular fatty
acids, but C16:1 ω9c was also observed only in strain HDW17BT
as the major cellular fatty acid. The respiratory quinone of the
isolates was MK-7. The major polar lipid components were
phosphatidylglycerol, phosphatidylethanolamine, and diphosphatidylglycerol.
The genomic DNA G + C content of strains
HDW17AT and HDW17BT were 36.6 and 34.4%, respectively.
Both strains had cell wall peptidoglycan composed of the
amino acids L-alanine, glycine, D-glutamic acid, L-tryptophan,
L-lysine, and L-aspartic acid, and the sugars ribose, glucose,
and galactose. Based on phylogenetic, phenotypic, chemotaxonomic,
and genotypic analyses, strains HDW17AT and
HDW17BT represent two novel species in the genus Vagococcus.
We propose the name Vagococcus coleopterorum sp.
nov. for strain HDW17AT (= KACC 21348T = KCTC 49324T
= JCM 33674T) and the name Vagococcus hydrophili sp. nov.
for strain HDW17BT (= KACC 21349T = KCTC 49325T =
JCM 33675T).
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The wide use of malachite green (MG) as a dye has caused
substantial concern owing to its toxicity. Bacillus cereus can
against the toxic effect of MG and efficiently decolourise it.
However, detailed information regarding its underlying adaptation
and degradation mechanisms based on proteomic
data is scarce. In this study, the isobaric tags for relative and
absolute quantitation (iTRAQ)-facilitated quantitative method
was applied to analyse the molecular mechanisms by
which B. cereus degrades MG. Based on this analysis, 209
upregulated proteins and 198 downregulated proteins were
identified with a false discovery rate of 1% or less during MG
biodegradation. Gene ontology and KEGG analysis determined
that the differentially expressed proteins were enriched
in metabolic processes, catalytic activity, antioxidant activity,
and responses to stimuli. Furthermore, real-time qPCR was
utilised to further confirm the regulated proteins involved
in benzoate degradation. The proteins BCE_4076 (Acetyl-CoA
acetyltransferase), BCE_5143 (Acetyl-CoA acetyltransferase),
BCE_5144 (3-hydroxyacyl-CoA dehydrogenase), BCE_4651
(Enoyl-CoA hydratase), and BCE_5474 (3-hydroxyacyl-CoA
dehydrogenase) involved in the benzoate degradation pathway
may play an important role in the biodegradation of MG
by B. cereus. The results of this study not only provide a comprehensive
view of proteomic changes in B. cereus upon MG
loading but also shed light on the mechanism underlying
MG biodegradation by B. cereus.
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Ogataea parapolymorpha (Hansenula polymorpha DL-1) is
a thermotolerant methylotrophic yeast with biotechnological
applications. Here, O. parapolymorpha genes whose expression
is induced in response to heat shock were identified by
transcriptome analysis and shown to possess heat shock elements
(HSEs) in their promoters. The function of O. parapolymorpha
HSF1 encoding a putative heat shock transcription
factor 1 (OpHsf1) was characterized in the context of heat
stress response. Despite exhibiting low sequence identity
(26%) to its Saccharomyces cerevisiae homolog, OpHsf1 harbors
conserved domains including a DNA binding domain
(DBD), domains involved in trimerization (TRI), transcriptional
activation (AR1, AR2), transcriptional repression (CE2),
and a C-terminal modulator (CTM) domain. OpHSF1 could
complement the temperature sensitive (Ts) phenotype of a
S. cerevisiae hsf1 mutant. An O. parapolymorpha strain with
an H221R mutation in the DBD domain of OpHsf1 exhibited
significantly retarded growth and a Ts phenotype. Intriguingly,
the expression of heat-shock-protein‒coding genes harboring
HSEs was significantly decreased in the H221R mutant
strain, even under non-stress conditions, indicating the importance
of the DBD for the basal growth of O. parapolymorpha.
Notably, even though the deletion of C-terminal domains
(ΔCE2, ΔAR2, ΔCTM) of OpHsf1 destroyed complementation
of the growth defect of the S. cerevisiae hsf1 strain,
the C-terminal domains were shown to be dispensable in O.
parapolymorpha. Overexpression of OpHsf1 in S. cerevisiae
increased resistance to transient heat shock, supporting the
idea that OpHsf1 could be useful in the development of heatshock‒
resistant yeast host strains.
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between Ganoderma boninense, a phytopathogenic fungus
that is particularly cytotoxic and pathogenic to plant tissues
and roots, and antimicrobial compounds. We previously observed
that liquid-liquid extraction (LLE) using chloroformmethanol-
water at a ratio (1:1:1) was superior at detecting
antibacterial activities and significant quantities of antibacterial
compounds. Herein, we demonstrate that antibacterial
secondary metabolites are produced from G. boninense mycelia.
Antibacterial compounds were monitored in concurrent
biochemical and biophysical experiments. The combined methods included high performance thin-layer chromatography
(HPTLC), gas chromatography-mass spectrometry
(GC-MS), high-performance liquid chromatography (HPLC),
fourier transform infrared (FTIR), and nuclear magnetic resonance
(NMR) spectroscopy. The antibacterial compounds
derived from mycelia with chloroform-methanol extraction
through LLE were isolated via a gradient solvent elution system
using HPTLC. The antibacterial activity of the isolated
compounds was observed to be the most potent against Staphylococcus
aureus ATCC 25923 and multidrug-resistant S.
aureus NCTC 11939. GC-MS, HPLC, and FTIR analysis confirmed
two antibacterial compounds, which were identified
as 4,4,14α-trimethylcholestane (m/z = 414.75; lanostane,
C30H54) and ergosta-5,7,22-trien-3β-ol (m/z = 396.65; ergosterol,
C28H44O). With the aid of spectroscopic evaluations,
ganoboninketal (m/z = 498.66, C30H42O6), which belongs to
the 3,4-seco-27-norlanostane triterpene family, was additionally
characterized by 2D-NMR analysis. Despite the lack of
antibacterial potential exhibited by lanostane; both ergosterol
and ganoboninketal displayed significant antibacterial activities
against bacterial pathogens. Results provide evidence
for the existence of bioactive compounds in the mycelia of
the relatively unexplored phytopathogenic G. boninense, together
with a robust method for estimating the corresponding
potent antibacterial secondary metabolites.
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Putrescine, a typical polyamine compound important for
cell growth and stress resistance, can be utilized as an energy
source. However, the regulation of its catabolism is unclear.
Here the small RNA (sRNA) Spot 42, an essential regulator
of carbon catabolite repression (CCR), was confirmed to participate
in the post-transcriptional regulation of putrescine
catabolism in Escherichia coli. Its encoding gene spf exclusively
exists in the γ-proteobacteria and contains specific binding
sites to the 5-untranslated regions of the puuE gene, which
encodes transaminase in the glutamylated putrescine pathway
of putrescine catabolism converting γ-aminobutyrate
(GABA) into succinate semialdehyde (SSA). The transcription
of the spf gene was induced by glucose, inhibited by putrescine,
and unaffected by PuuR, the repressor of puu genes.
Excess Spot 42 repressed the expression of PuuE significantly
in an antisense mechanism through the direct and specific
base-pairing between the 51–57 nt of Spot 42 and the 5-
UTR of puuE. Interestingly, Spot 42 mainly influenced the
stability of the puuCBE transcript. This work revealed the regulatory
role of Spot 42 in putrescine catabolism, in the switch
between favorable and non-favorable carbon source utilization,
and in the balance of metabolism of carbon and nitrogen
sources.
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In prokaryotes, toxin-antitoxin (TA) systems are commonly
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or extremophiles to various unfavorable environments
by slowing their growth rate. Genomic analysis of the extremophile
Deinococcus radiodurans R1 revealed the presence of
eight type II TA systems, including the genes dr0417, dr0660,
dr1530, dr0690, and dr1807. Expression of these toxin genes
led to inhibition of Escherichia coli growth, whereas their
antidote antitoxins were able to recover the growth defect.
Remarkably, Dr0417 (DrMazF) showed endoribonuclease activity
toward rRNAs as well as mRNAs, as determined by in
vivo and in vitro RNA cleavage assays, and this activity was
inhibited by Dr0416 (DrMazE). It was also found that the expression
of dr0416-0417 module is directly regulated by the
DrMazE-MazF complex. Furthermore, this TA module was
induced under stress conditions and plays an important role
in survival. To understand the regulatory mechanism at the
molecular level, we determined the first high-resolution structures
of DrMazF alone and of the DrMazE-MazF complex.
In contrast with the hetero-hexameric state of typical MazEMazF
complexes found in other species, DrMazE-MazF crystal
structure consists of a hetero-trimer, with the DNA-binding
domain of DrMazE undergoing self-cleavage at the flexible
linker loop. Our structure revealed that the unique residue
R54 provides an additional positive charge to the substratebinding
pocket of DrMazF, its mutation significantly affects
the endonuclease activity. Thus, our work reports the unique
structural and biochemical features of the DrMazE-MazF
system.
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5-Fluorouracil (5-FU) is an essential drug in systemic chemotherapy
treatments for colorectal cancer (CRC). Despite
the development of several treatment strategies over the past
decades, the patient benefits of 5-FU-based therapies have
been compromised by the development of chemoresistance.
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be due to genetic and epigenetic factors unique to individuals.
Therefore, important factors for realizing personalized medicine
are to accurately understand the causes and mechanisms
of drug resistance to 5-FU-based therapies and to identify
and validate prognostic biomarkers. Gut microbes that
interact directly with the host contribute to human health
and cancer control. Lactobacillus plantarum, in particular, has
the potential to be a therapeutic agent by producing bioactive
compounds that may benefit the host. Here, we investigated
the gamma-aminobutyric acid (GABA) and GABAB
receptor (GABABR)-dependent signaling pathway as a treatment
option for 5-FU-resistant HT-29 cells. GABA-producing
L. plantarum activates anti-proliferative, anti-migration,
and anti-invasion effects against 5-FU-resistant HT-29 cells.
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mediated via GABABR. Activated GABABR induces apoptosis
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pathways and cellular inhibitor of apoptosis protein 2 (cIAP2)
expression. Thus, the GABAergic system has potential in 5-
FU-resistant HT-29 cells as a predictive biomarker. In addition,
GABA-producing L. plantarum is promising as an adjuvant
treatment for 5-FU-resistant CRC, and its intervention
in neurobiological signaling imply new possibilities for
chemoprevention and the treatment of colon cancer-related
diseases.
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In vitro fecal fermentation is an assay that uses fecal microbes
to ferment foods, the results of which can be used to
evaluate the potential of prebiotic candidates. To date, there
have been various protocols used for in vitro fecal fermentation-
based assessments of food substances. In this study,
we investigated how personal gut microbiota differences and
external factors affect the results of in vitro fecal fermentation
assays. We used Cheonggukjang (CGJ), a Korean traditional
fermented soybean soup that is acknowledged as
healthy functional diet. CGJ was digested in vitro using acids
and enzymes, and then fermented with human feces anaerobically.
After fecal fermentation, the microbiota was analyzed
using MiSeq, and the amount of short chain fatty acids
(SCFAs) were measured using GC-MS. Our results suggest
that CGJ was effectively metabolized by fecal bacteria to produce
SCFAs, and this process resulted in an increase in the
abundance of Coprococcus, Ruminococcus, and Bifidobacterium
and a reduction in the growth of Sutterella, an opportunistic
pathogen. The metabolic activities predicted from the
microbiota shifts indicated enhanced metabolism linked to
methionine biosynthesis and depleted chondroitin sulfate
degradation. Moreover, the amount of SCFAs and microbiota
shifts varied depending on personal microbiota differences.
Our findings also suggest that in vitro fecal fermentation of
CGJ for longer durations may partially affect certain fecal
microbes. Overall, the study discusses the usability of in vitro
gastrointestinal digestion and fecal fermentation (GIDFF)
to imitate the effects of diet-induced microbiome modulation
and its impact on the host.
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