Biofilms are complex microbial architectures that attach to
surfaces and encase microorganisms in a matrix composed
of self-produced hydrated extracellular polymeric substances
(EPSs). In biofilms, microorganisms become much more
resistant to antimicrobial treatments, harsh environmental
conditions, and host immunity. Biofilm formation by microbial
pathogens greatly enhances survival in hosts and causes
chronic infections that result in persistent inflammation and
tissue damages. Currently, it is believed over 80% of chronic
infectious diseases are mediated by biofilms, and it is known
that conventional antibiotic medications are inadequate at
eradicating these biofilm-mediated infections. This situation
demands new strategies for biofilm-associated infections,
and currently, researchers focus on the development of antibiofilm
agents that are specific to biofilms, but are nontoxic,
because it is believed that this prevents the development of
drug resistance. Here, we review the most promising antibiofilm
agents undergoing intensive research and development.
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Gram-staining-negative, uniflagellated, rod-shaped, designated
as DCY110T, was isolated from sludge located in Gangwon
province, Republic of Korea. The phylogenetic tree of 16S
rRNA gene sequence showed that the strain DCY110T belonged
to the genus Rhodoferax with a close similarity to Rhodoferax
saidenbachensis DSM 22694T (97.7%), Rhodoferax
antarcticus DSM 24876T (97.5%), Rhodoferax ferrireducens
DSM 15236T (97.3%), and Rhodoferax fermentans JCM 7819T
(96.7%). The predominant isoprenoid quinine was ubiquinone
(Q-8). DNA G + C content was 62.8 mol%. The major
polar lipids were phosphatidylethanolamine and two unidentified
phospholipids. The major fatty acids (> 10%) were
C12:0, C16:0, summed feature 3 (which comprised C16:1 ω7c
and/or C16:1 ω6c). The DNA-DNA relatedness values between
the strain DCY110T and the closely related relatives used in
this study were lower than 70%. Based on the following polyphasic
analysis, the strain DCY110T is considered as a novel
species of the genus Rhodoferax, for which the name Rhodoferax
koreense sp. nov. is proposed. The type strain is DCY-
110T (= KCTC 52288T = JCM 31441T).
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that was isolated from a freshwater lake using the floating
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16S rRNA gene sequences, the isolate was found to be closely
related to the genus Methylomonas in the family Methylococcaceae
of the class Gammaproteobacteria with 94.2–97.4%
16S rRNA gene similarity to Methylomonas type strains. Comparison
of chemotaxonomic and physiological properties
further suggested that strain EMGL16-1 was taxonomically
distinct from other species in the genus Methylomonas. The
isolate was versatile in utilizing nitrogen sources such as molecular
nitrogen, nitrate, nitrite, urea, and ammonium. The
genes coding for subunit of the particulate form methane
monooxygenase (pmoA), soluble methane monooxygenase
(mmoX), and methanol dehydrogenase (mxaF) were detected
in strain EMGL16-1. Phylogenetic analysis of mmoX indicated
that mmoX of strain EMGL16-1 is distinct from those
of other strains in the genus Methylomonas. This isolate probably
represents a novel species in the genus. Our study provides
new insights into the diversity of species in the genus
Methylomonas and their environmental adaptations.
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The 25 kDa subunit of the Clevage Factor Im (CFIm25) is
an essential factor for messenger RNA polyadenylation in
human cells. Therefore, here we investigated whether the homologous
protein of Entamoeba histolytica, the protozoan
responsible for human amoebiasis, might be considered as
a biochemical target for parasite control. Trophozoites were
cultured with bacterial double-stranded RNA molecules targeting
the EhCFIm25 gene, and inhibition of mRNA and protein
expression was confirmed by RT-PCR and Western blot
assays, respectively. EhCFIm25 silencing was associated with
a significant acceleration of cell proliferation and cell death.
Moreover, trophozoites appeared as larger and multinucleated
cells. These morphological changes were accompanied by a
reduced mobility, and erythrophagocytosis was significantly
diminished. Lastly, the knockdown of EhCFIm25 affected the
poly(A) site selection in two reporter genes and revealed that
EhCFIm25 stimulates the utilization of downstream poly(A)
sites in E. histolytica mRNA. Overall, our data confirm that
targeting the polyadenylation process represents an interesting
strategy for controlling parasites, including E. histolytica.
To our best knowledge, the present study is the first to
have revealed the relevance of the cleavage factor CFIm25
as a biochemical target in parasites.
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is cultivated widely in China. In this study, a genetic linkage
map for A. auricula-judae was constructed using a mapping
population consisting of 138 monokaryons derived from a
hybrid strain (A119-5). The monokaryotic parent strains
A14-5 and A18-119 were derived from two cultivated varieties,
A14 (Qihei No. 1) and A18 (Qihei No. 2), respectively.
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of A. auricula-judae and amplified in A14-5, A18-
119, and the mapping population. The map consisted of 11
linkage groups (LGs) spanning 854 cM, with an average interval
length of 6.57 cM. A testcross population was derived
from crossing between the monokaryon A184-57 (from the
wild strain A184 as a tester strain) and the mapping population.
Important agronomic trait-related QTLs, including
mycelium growth rate on potato dextrose agar for the mapping
population, mycelium growth rate on potato dextrose
agar and sawdust for the testcross population, growth period
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for the testcross population, were identified using the composite
interval mapping method. Six mycelium growth raterelated
QTLs were identified on LG1 and LG4, two growth
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QTLs were identified on LG2 and LG6. The results
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for locating genes for important agronomic characteristics
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a Burkholderia lata isolate from a pig with swine respiratory
disease in Korea was screened for strains showing attenuated
virulence in Caenorhabditis elegans. One such mutant was
obtained, and the Tn5 insertion junction was mapped to
rpfR, a gene encoding a cyclic di-GMP phosphodiesterase that
functions as a receptor. Mutation of rpfR caused a reduction
in growth on CPG agar and swimming motility as well as a
rough colony morphology on Congo red agar. TLC analysis
showed reduced AHL secretion, which was in agreement with
the results from plate-based and bioluminescence assays. The
mutant strain produced significantly more biofilm detected
by crystal violet staining than the parent strain. SEM of the
mutant strain clearly showed that the overproduced biofilm
contained a filamentous structure. These results suggest
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field (EMF) from the devices is a potential health
threat. Although the direct health effect of a cell phone and its
radiofrequency (RF) EMF to human is still elusive, the effect
to unicellular organisms is rather apparent. Human microbiota,
including skin microbiota, has been linked to a very
significant role in the health of a host human body. It is important
to understand the response of human skin microbiota
to the RF-EMF from cell phones and personal electronic
devices, since this may be one of the potential mechanisms
of a human health threat brought about by the disruption
of the intimate and balanced host-microbiota relationship.
Here, we investigated the response of both laboratory culture
strains and isolates of skin bacteria under static magnetic
field (SMF) and RF-EMF. The growth patterns of laboratory
cultures of Escherichia coli, Pseudomonas aeruginosa,
and Staphylococcus epidermidis under SMF were variable
per different species. The bacterial isolates of skin microbiota
from 4 subjects with different cell phone usage history also
showed inconsistent growth responses. These findings led us
to hypothesize that cell phone level RF-EMF disrupts human
skin microbiota. Thus, the results from the current study lay
ground for more comprehensive research on the effect of
RF-EMF on human health through the human-microbiota
relationship.
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The Escherichia coli cAMP receptor protein (CRP) utilizes the
helix-turn-helix motif for DNA binding. The CRP’s recognition
helix, termed F-helix, includes a stretch of six amino
acids (Arg180, Glu181, Thr182, Val183, Gly184, and Arg185)
for direct DNA contacts. Arg180, Glu181 and Arg185 are
known as important residues for DNA binding and specificity,
but little has been studied for the other residues. Here
we show that Gly184 is another F-helix residue critical for
the transcriptional activation function of CRP. First, glycine
was repeatedly selected at CRP position 184 for its unique
ability to provide wild type-level transcriptional activation
activity. To dissect the glycine requirement, wild type CRP
and mutants G184A, G184F, G184S, and G184Y were purified
and their in vitro DNA-binding activity was measured.
G184A and G184F displayed reduced DNA binding, which
may explain their low transcriptional activation activity. However,
G184S and G184Y displayed apparently normal DNA
affinity. Therefore, an additional factor is needed to account
for the diminished transcriptional activation function in
G184S and G184Y, and the best explanation is perturbations
in their interaction with RNA polymerase. The fact that glycine
is the smallest amino acid could not fully warrant its suitability,
as shown in this study. We hypothesize that Gly184
fulfills the dual functions of DNA binding and RNA polymerase
interaction by conferring conformational flexibility
to the F-helix.
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for more than 30 years. Our previous studies confirmed
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can protect mice against nasopharyngeal colonization of S.
pneumoniae and lethal pneumococcal infection, and the
protective effects are comparable with those induced by commercially
available 23-valent polysaccharide vaccine. However,
live attenuated vaccine SPY1 needs four inoculations to
get satisfactory protective effect, which may increase the risk
of virulence recovery. It is reported that heterologous primeboost
approach is more effective than homologous primeboost
approach. In the present study, to decrease the doses
of live SPY1 and improve the safety of SPY1 vaccine, we immunized
mice with SPY1 and DnaJ protein alternately. Our results showed that heterologous prime-boost immunization
with SPY1 and DnaJ protein could significantly reduce
the colonization of S. pneumoniae in the respiratory tract of
mice, and induce stronger Th1 and Th17 cellular immune
responses than SPY1 alone. These results indicate heterologous
prime-boost immunization method not only elicits
better protective effect than SPY1 alone, but also reduces the
doses of live SPY1 and decreases the risk of SPY1 vaccine.
This work is the first time to study the protective efficiency
with two different forms of S. pneumoniae candidate vaccine,
and provides a new strategy for the development of S. pneumoniae
vaccine.
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Streptococcus mutans is a major etiologic agent of human
dental caries that forms biofilms on hard tissues in the human
oral cavity, such as tooth and dentinal surfaces. Human
β-defensin-3 (HBD3) is a 45-amino-acid natural antimicrobial
peptide that has broad spectrum antimicrobial activity
against bacteria and fungi. A synthetic peptide consisting of
the C-terminal 15 amino acids of HBD3 (HBD3-C15) was
recently shown to be sufficient for its antimicrobial activity.
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attention. In this study, we investigated whether HBD3-C15
inhibits the growth of the representative cariogenic pathogen
Streptococcus mutans and its biofilm formation. HBD3-C15
inhibited bacterial growth, exhibited bactericidal activity,
and attenuated bacterial biofilm formation in a dose-dependent
manner. HBD3-C15 potentiated the bactericidal and
anti-biofilm activity of calcium hydroxide (CH) and chlorhexidine
digluconate (CHX), which are representative disinfectants
used in dental clinics, against S. mutans. Moreover,
HBD3-C15 showed antimicrobial activity by inhibiting biofilm
formation by S. mutans and other dentinophilic bacteria
such as Enterococcus faecalis and Streptococcus gordonii,
which are associated with dental caries and endodontic
infection, on human dentin slices. These effects were observed
for HBD3-C15 alone and for HBD3-C15 in combination with
CH or CHX. Therefore, we suggest that HBD3-C15 is a potential
alternative or additive disinfectant that can be used
for the treatment of oral infectious diseases, including dental
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