During meiosis, crossing over allows for the exchange of genes
between homologous chromosomes, enabling their segregation
and leading to genetic variation in the resulting gametes.
Spo11, a topoisomerase-like protein expressed in eukaryotes,
and diverse accessory factors induce programmed doublestrand
breaks (DSBs) to initiate meiotic recombination during
the early phase of meiosis after DNA replication. DSBs
are further repaired via meiosis-specific homologous recombination.
Studies on budding yeast have provided insights
into meiosis and genetic recombination and have improved
our understanding of higher eukaryotic systems. Cohesin, a
chromosome-associated multiprotein complex, mediates sister
chromatid cohesion (SCC), and is conserved from yeast
to humans. Diverse cohesin subunits in budding yeast have
been identified in DNA metabolic pathways, such as DNA
replication, chromosome segregation, recombination, DNA
repair, and gene regulation. During cell cycle, SCC is established
by multiple cohesin subunits, which physically bind
sister chromatids together and modulate proteins that involve
in the capturing and separation of sister chromatids. Cohesin
components include at least four core subunits that establish
and maintain SCC: two structural maintenance chromosome
subunits (Smc1 and Smc3), an α-kleisin subunit (Mcd1/Scc1
during mitosis and Rec8 during meiosis), and Scc3/Irr1 (SA1
and SA2). In addition, the cohesin-associated factors Pds5
and Rad61 regulate structural modifications and cell cyclespecific
dynamics of chromatin to ensure accurate chromosome
segregation. In this review, we discuss SCC and the
recombination pathway, as well as the relationship between
the two processes in budding yeast, and we suggest a possible
conserved mechanism for meiotic chromosome dynamics
from yeast to humans.
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A Gram-stain-negative, strictly aerobic bacterial strain, designated
strain S27T, was isolated from soil near an artificial
pond in South Korea. Cells were non-motile short rods showing
oxidase- and catalase-positive activities. Growth of strain
S27T was observed at 20–40°C (optimum, 30°C), pH 5.0–7.0
(optimum, pH 6.0), and 0–0.5% (w/v) NaCl (optimum, 0%).
Ubiquinone-8 was detected as the sole respiratory quinone
and the major fatty acids were C16:0, cyclo-C17:0, and cyclo-
C19:0 ω8c. The G + C content of the genomic DNA was 62.4
mol%. Phosphatidylglycerol, phosphatidylethanolamine, and
an unidentified aminophospholipid were detected as the major
polar lipids. Phylogenetic analysis based on 16S rRNA gene
sequences showed that strain S27T formed a clearly distinct
phyletic lineage from closely related Paraburkholderia species
within the genus Paraburkholderia. Strain S27T was most
closely related to Paraburkholderia rhynchosiae WSM3937T,
Paraburkholderia ginsengiterrae DCY85T, Paraburkholderia
fungorum NBRC 102489T, and Paraburkholderia graminis
C4D1MT with 98.8%, 98.4%, 98.4%, and 97.7% 16S rRNA
gene sequence similarities, respectively. The DNA-DNA relatedness
level between strain S27T and the type strain of P.
rhynchosiae was 36.8 ± 2.6%. On the basis of phenotypic, chemotaxonomic
and molecular properties, strain S27T represents
a novel species of the genus Paraburkholderia, for which
the name Paraburkholderia lacunae sp. nov. is proposed. The
type strain is S27T (KACC 19714 T = JCM 32721T).
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A Gram-stain-negative, non-spore-forming, facultative, rodshaped
bacterium (designated LA-28T) was isolated from a
sludge sample from a wastewater treatment plant in Hanam
city, Republic of Korea. On the basis of 16S rRNA gene sequencing,
strain LA-28T clustered with species of the genus
Mesorhizobium and appeared closely related to M. jarvisii
LMG 28313T (96.8%), M. waimense ICMP 19557T (96.7%),
and M. huakuii LMG 14107T (96.7%). Growth occurs at 18–
40°C on R2A medium in the presence of 1–4% NaCl (w/v)
and at pH 6–8. The DNA G+C content was 61.2 mol%, and
the predominant quinone was ubiquinone-10 (Q-10). The
major cellular fatty acids (> 5%) were C16:0, C19:0 ω8c cyclo,
C18:1 ω7c 11-methyl, and C18:1 ω7c and/or C18:1 ω6c (summed
feature 8). Major polar lipids were phosphatidylglycerol (PG),
phosphatidylethanolamine (PE), phosphatidyl-N-methylethanolamine
(PME), and phosphatidylcholine (PC). Physiological
and biochemical characteristics indicated that strain
LA-28T represents a novel species of the genus Mesorhizobium,
for which the name Mesorhizobium denitrificans sp.
nov. is proposed. The type strain is LA-28T (= KACC 19675T
= LMG 30806T).
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Various Nostoc spp. and related cyanobacteria are able to survive
extreme temperatures and are among the most successful
colonists of high-elevation sites being exposed due to glacial
retreat. It is unclear, however, if cyanobacteria can grow
during the extreme freeze-thaw cycles that occur on a yearround
basis at high-elevation, peri-glacial sites or if they only
grow during the rare periods when freeze-thaw cycles do not
occur. We conducted several experiments to determine if cyanobacteria
that form biological soil crusts (BSCs) at highelevation
sites (> 5,000 m.a.s.l.) in the Andes can grow during
diurnal freeze-thaw cycles on a par with those that occur in
the field. Here we show that a soil crust that had been frozen
at -20°C for five years was able to increase from 40% to 100%
soil coverage during a 45-day incubation during which the
soil temperature cycled between -12°C and 26°C every day.
In a second, experiment an undeveloped soil with no visible
BSCs showed a statistically significant shift in the bacterial
community from one containing few cyanobacterial sequences
(8% of sequences) to one dominated (27%) by Nostoc,
Microcoleus, and Leptolyngbya phylotypes during a 77-day
incubation with daily freeze-thaw cycles. In addition, counts
of spherical Nostoc-like colonies increased significantly on
the soil surface during the experiment, especially in microcosms
receiving phosphorus. Taken together these results
show that freeze-thaw cycles alone do not limit the growth
of BSCs in high-elevation soils, and provide new insight into
how life is able to thrive in one of the most extreme terrestrial
environments on Earth.
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year. Associations of bacteria with diatoms, green microalgae,
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Since molecular genotyping has been established for the
Candida species, studies have found that a single Candida
strain (endemic strain) can persist over a long period of time
and results in the spread of nosocomial invasive candidiasis
without general characteristics of horizontal transmissions.
Our previous study also found the existence of endemic
strains in a cancer center in Tianjin, China. In the current
study, we performed further investigation on endemic and
non-endemic Candida albicans strains, with the aim of explaining
the higher morbidity of endemic strains. In an in
vivo experiment, mice infected with endemic strains showed
significantly shorter survival time and higher kidney fungal
burdens compared to mice infected with non-endemic strains.
In an in vitro experiment, the killing percentage of neutrophils
to endemic strains was significantly lower than that to
non-endemic strains, which is positively linked to the ratio
of LC3B-II/I in neutrophils. An immunofluorescence assay
showed more β-1,3-glucan exposure on the cell walls of nonendemic
strains compared to endemic strains. After blocking
the β-glucan receptor (CR3) or inhibiting downstream
kinase (SYK) in neutrophils, the killing percent to C. albicans
(regardless of endemic and non-endemic strains) and the ratio
of LC3B-II/I of neutrophils were significantly decreased.
These data suggested that the killing capability of neutrophils
to C. albicans was monitored by β-1,3-glucan via CR3/SYK
pathway-dependent LC3B-II accumulation and provided
an explanation for the variable killing capability of neutrophils
to different strains of C. albicans, which would be beneficial
in improving infection control and therapeutic strategies
for invasive candidiasis.
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Multi-drug resistant (MDR) non-typhoidal Salmonella (NTS)
is increasingly common worldwide. While food animals are
thought to contribute to the growing antimicrobial resistance
(AMR) problem, limited data is documenting this relationship,
especially in low and middle-income countries (LMIC).
Herein, we aimed to assess the role of non-clinical NTS of bovine
origin as reservoirs of AMR genes of human clinical significance.
We evaluated the phenotypic and genotypic AMR
profiles in a set of 44 bovine-associated NTS. For comparative
purposes, we also included genotypic AMR data of additional
isolates from Mexico (n = 1,067) that are publicly available.
The most frequent AMR phenotypes in our isolates involved
tetracycline (40/44), trimethoprim-sulfamethoxazole (26/44),
chloramphenicol (19/44), ampicillin (18/44), streptomycin
(16/44), and carbenicillin (13/44), while nearly 70% of the
strains were MDR. These phenotypes were correlated with
a widespread distribution of AMR genes (i.e. tetA, aadA,
dfrA12, dfrA17, sul1, sul2, bla-TEM-1, blaCARB-2) against
multiple antibiotic classes, with some of them contributed by
plasmids and/or class-1 integrons. We observed different
AMR genotypes for betalactams and tetracycline resistance,
providing evidence of convergent evolution and adaptive AMR.
The probability of MDR genotype occurrence was higher in
meat-associated isolates than in those from other sources (odds
ratio 11.2, 95% confidence interval 4.5–27.9, P < 0.0001). The
study shows that beef cattle are a significant source of MDR NTS in Mexico, highlighting the role of animal production
on the emergence and spread of MDR Salmonella in LMIC.
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Poly(β-L-malic acid) (PMA) is a promising polyester formed
from L-malate in microbial cells. Malate biosynthesis is crucial
for PMA production. Previous studies have shown that
the non-oxidative pathway or oxidative pathway (TCA cycle)
is the main biosynthetic pathway of malate in most of PMAproducing
strains, while the glyoxylate cycle is only a supplementary
pathway. In this study, we investigated the effect
of exogenous metabolic intermediates and inhibitors of the
malate biosynthetic pathway on PMA production by Aureobasidium
melanogenum GXZ-6. The results showed that PMA
production was stimulated by maleic acid (a fumarase inhibitor)
and sodium malonate (a succinate dehydrogenase inhibitor)
but inhibited by succinic acid and fumaric acid. This
indicated that the TCA cycle might not be the only biosynthetic
pathway of malate. In addition, the PMA titer increased
by 18.1% upon the addition of glyoxylic acid after 72 h of fermentation,
but the PMA titer decreased by 7.5% when itaconic
acid (an isocitrate lyase inhibitor) was added, which indicated
that malate for PMA production was synthesized significantly
via the glyoxylate cycle rather than the TCA cycle. Furthermore,
in vitro enzyme activities of the TCA and glyoxylate
cycles suggested that the glyoxylate cycle significantly contributed
to the PMA production, which is contradictory to what
has been reported previously in other PMA-producing A.
pullulans.
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The detailed antibacterial mechanism of cordycepin efficacy
against food-borne germs remains ambiguous. In this study,
the antibacterial activity and action mechanism of cordycepin
were assessed. The results showed that cordycepin effectively
inhibited the growth of seven bacterial pathogens
including both Gram-positive and Gram-negative bacterial
pathogens; the minimum inhibitory concentrations (MIC)
were 2.5 and 1.25 mg/ml against Escherichia coli and Bacillus
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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Among the major enteric pathogens, Campylobacter jejuni is
considered an important source of diarrheal illness in humans.
In contrast to the acute gastroenteritis in humans, C. jejuni
exhibits prolonged cecal colonization at a high level with little
or no pathology in chickens. Although several known virulence
determinants of C. jejuni have been found to be associated
with a higher degree of pathogenesis in humans, to date, little
is known about their functions in the persistent colonization
of chickens. The present study was undertaken to assess the
role of C. jejuni in imparting differential host immune responses
in human and chicken cells. Based on the abundance
of major genes encoding virulence factors (GEVFs), we used
a particular isolate that harbors the cadF, flaA, peb1, racR,
ciaB, cdtB, and hcp genes. This study showed that hypervirulent
C. jejuni isolate that encodes a functional type VI secretion
system (T6SS) has a greater ability to invade and create
characteristic “attaching and effacing” lesions in human
INT407 compared to primary chicken embryo intestinal cells
(CEICs). Furthermore, we demonstrated that the higher bacterial
invasion in human INT407 triggered higher levels of
expression of major proinflammatory cytokines, such as IL-
1β and IL-6, and significant downregulation of IL-17A gene
expression (P ≤ 0.05). The findings of the present study suggest
that the enhanced ability of C. jejuni to invade human
cells is tightly regulated by proinflammatory cytokines in the
gut and possibly holds the keys to the observed differences
in pathogenesis between human and chicken cells.
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Immunopathological properties of the Campylobacter jejuni flagellins and the adhesin CadF as assessed in a clinical murine infection model Anna-Maria Schmidt, Ulrike Escher, Soraya Mousavi, Nicole Tegtmeyer, Manja Boehm, Steffen Backert, Stefan Bereswill, Markus M. Heimesaat Gut Pathogens.2019;[Epub] CrossRef
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Enterococcus faecalis, a Gram-positive bacterium commonly
isolated in patients with refractory apical periodontitis, invades
dentin tubules easily and forms biofilms. Bacteria in biofilms,
which contribute to recurrent and/or chronic inflammatory
diseases, are more resistant to antimicrobial agents
than planktonic cells and easily avoid phagocytosis. Although
Lactobacillus plantarum lipoteichoic acid (Lp.LTA) is associated
with biofilm formation, the effect of Lp.LTA on biofilm
formation by E. faecalis is not clearly understood. In this
study, we investigated whether Lp.LTA inhibits E. faecalis
biofilm formation. The degree of biofilm formation was determined
by using crystal violet assay and LIVE/DEAD bacteria
staining. The quantification of bacterial growth was determined
by measuring the optical density at 600 nm with a
spectrophotometer. Formation of biofilms on human dentin
slices was observed under a scanning electron microscope.
E. faecalis biofilm formation was reduced by Lp.LTA treatment
in a dose-dependent manner. Lp.LTA inhibited biofilm
development of E. faecalis at the early stage without affecting
bacterial growth. LTA from other Lactobacillus species
such as Lactobacillus acidophilus, Lactobacillus casei, or
Lactobacillus rhamnosus GG also inhibited E. faecalis biofilm
formation. In particular, among LTAs from various lactobacilli,
Lp.LTA showed the highest inhibitory effect on biofilms
formed by E. faecalis. Interestingly, LTAs from lactobacilli
could remove the biofilm preformed by E. faecalis.
These inhibitory effects were also observed on the surface of human dentin slices. In conclusion, Lactobacillus species LTA
inhibits biofilm formation caused by E. faecalis and it could
be used as an anti-biofilm agent for prevention or treatment
against E. faecalis-associated diseases.
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African swine fever (ASF) is a highly lethal haemorrhagic
disease in domestic and wild swine that has acquired great
importance in sub-Saharan Africa since 1997. ASF was first
reported in Cameroon in 1982 and was detected only in Southern
Cameroon (South, West, East, Northwest, Southwest,
Littoral, and Centre regions) until February 2010 when suspected
ASF outbreaks were reported in the North and Far
North regions. We investigated those outbreaks by analysing
samples that were collected from sick pigs between 2010 and
2018. We confirmed 428 positive samples by ELISA and realtime
PCR and molecularly characterized 48 representative
isolates. All the identified virus isolates were classified as ASFV
genotype I based on the partial B646L gene (C-terminal end
of VP72 gene) and the full E183L gene encoding p54 protein
analysis. Furthermore, analysis of the central variable region
(CVR) within the B602L gene demonstrated that there were
3 different variants of ASFV genotype I, with 19, 20, and 21
tetrameric tandem repeat sequences (TRSs), that were involved
in the 2010–2018 outbreaks in Cameroon. Among
them, only variant A (19 TRSs) was identical to the Cam/82
isolate found in the country during the first outbreaks in 1981–1982. This study demonstrated that the three variants
of ASFV isolates involved in these outbreaks were similar to
those of neighbouring countries, suggesting a movement of
ASFV strains across borders. Designing common control
measures in affected regions and providing a compensation
programme for farmers will help reduce the incidence and
spread of this disease.
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