scholarly journals High Prevalence of Diverse Insertion Sequences within the rRNA Operons of Mycoplasma bovis

2016 ◽  
Vol 82 (21) ◽  
pp. 6386-6394 ◽  
Author(s):  
Eytan Amram ◽  
Ilya Borovok ◽  
Yaarit Nachum-Biala ◽  
Roger Ayling ◽  
Uri Lerner ◽  
...  
Keyword(s):  
Pcr Analysis ◽  
Mycoplasma Bovis ◽  
Insertion Sequences ◽  
Putative Promoter ◽  
Ribosome Synthesis ◽  
Content Type ◽  
Is Elements ◽  
Viable Cells ◽  
High Prevalence ◽  
And Function

ABSTRACTInsertion sequences (ISs) are widespread in the genome ofMycoplasma bovisstrain PG45, but no ISs were identified within its two tandemly positioned rRNA operons (rrn1andrrn2). However, characterization of therrnlocus in 70M. bovisisolates revealed the presence of ISs related to the ISMbov1(IS30family) and ISMbov4(IS4family) isomers in 35 isolates. ISs were inserted into intergenic region 1 (IGR-1) or IGR-3, which are the putative promoter regions ofrrn1andrrn2, respectively, and into IGR-5, located downstream of therrl2gene. Seven different configurations (A to G) of therrnlocus with respect to ISs were identified, including those in five annotated genomes. The transcriptional start site for the singlerrnoperon inM. bovisstrain 88127 was mapped within IGR-1, 60 bp upstream of therrsgene. Notably, only 1 nucleotide separated the direct repeat (DR) for ISMbov1and the promoter –35 element in configuration D, while in configuration F, the −35 motif was a part of the ISMbov1DR. Relative quantitative real-time (qRT) PCR analysis and growth rate comparisons detected a significant increase (P< 0.05) in the expression of therrsgenes and in the number of viable cells during log phase growth (8, 12, and 16 h) in the strains with configuration F in comparison to strains with one or tworrnoperons that did not have ISs. A high prevalence of IS elements within or close to theM. bovisrrnoperon-promoter region may reflect their important role in regulation of both ribosome synthesis and function.IMPORTANCEData presented in this study show a high prevalence of diverse ISs within theM. bovis rrnlocus resulting in intraspecies variability and diversity. Such abundance of IS elements near or within therrnlocus may offer a selective advantage toM. bovis. Moreover, the fact that expression of therrsgenes as well as the number of viable cells increased in the group of strains with IS element insertion within a putative promoter −35 sequence (configuration F) in comparison to that in strains with one or tworrnoperons that do not have ISs may serve as a basis for understanding the possible role ofM. bovisIS elements in fundamental biological processes such as regulation of ribosome synthesis and function.

scholarly journals Molecular Investigation and Phylogeny of Anaplasma spp. in Mediterranean Ruminants Reveal the Presence of Neutrophil-Tropic Strains Closely Related to A. platys

2013 ◽  
Vol 80 (1) ◽  
pp. 271-280 ◽  
Author(s):  
Rosanna Zobba ◽  
Antonio G. Anfossi ◽  
Maria Luisa Pinna Parpaglia ◽  
Gian Mario Dore ◽  
Bernardo Chessa ◽  
...  
Keyword(s):  
Molecular Typing ◽  
Evolutionary History ◽  
Pcr Analysis ◽  
Serological Tests ◽  
Content Type ◽  
High Prevalence ◽  
Molecular Investigation ◽  
History Of ◽  
Few Data ◽  
Representative Area

ABSTRACTFew data are available on the prevalence and molecular typing of species belonging to the genusAnaplasmain Mediterranean ruminants. In this study, PCR analysis and sequencing of both 16S rRNA andgroELgenes were combined to investigate the presence, prevalence, and molecular traits ofAnaplasmaspp. in ruminants sampled on the Island of Sardinia, chosen as a subtropical representative area. The results demonstrate a high prevalence ofAnaplasmaspp. in ruminants, with animals infected by at least four of sixAnaplasmaspecies (Anaplasma marginale,A. bovis,A. ovis, andA. phagocytophilum). Moreover, ruminants host a number of neutrophil-tropic strains genetically closely related to the canine pathogenA. platys. The highAnaplasmaspp. prevalence and the identification of as-yet-unclassified neutrophil-tropic strains raise concerns about the specificity of serological tests routinely used in ruminants and provide additional background for reconstructing the evolutionary history of species genetically related toA. phagocytophilum.

scholarly journals Molecular Epidemiology and Characterization of Carbapenem-Resistant Klebsiella pneumoniae Isolated from Urine at a Teaching Hospital in Taiwan

2021 ◽  
Vol 9 (2) ◽  
pp. 271
Author(s):  
Yuarn-Jang Lee ◽  
Chih-Hung Huang ◽  
Noor Andryan Ilsan ◽  
I-Hui Lee ◽  
Tzu-Wen Huang
Keyword(s):  
Klebsiella Pneumoniae ◽  
Teaching Hospital ◽  
Efflux Pump ◽  
Resistance Mechanisms ◽  
Pcr Analysis ◽  
Carbapenem Resistant ◽  
High Prevalence ◽  
Antimicrobial Susceptibility Tests ◽  
Susceptibility Tests ◽  
Tract Infections

Urinary tract infections (UTIs) are common in clinics and hospitals and are associated with a high economic burden. Enterobacterium Klebsiella pneumoniae is a prevalent agent causing UTIs. A high prevalence of carbapenem-resistant K. pneumoniae (CRKP) has emerged recently and is continuing to increase. Seventeen urinary CRKP isolates collected at a teaching hospital in Taiwan from December 2016 to September 2017 were analyzed to elucidate their drug resistance mechanisms. Two-thirds of the isolates were obtained from outpatients. Antimicrobial susceptibility tests demonstrated multidrug resistance in all the isolates. Multilocus sequence typing analysis showed high diversity among the isolates. PCR analysis demonstrated the presence of carbapenemases in three isolates. All isolates carried at least one other extended-spectrum β-lactamase, including TEM, DHA, and CTX-M. Fifteen isolates contained mutations in one of the outer membrane porins that were assessed. The expression levels of the acrB and/or oqxB efflux pump genes, as determined by qRT-PCR, were upregulated in 11 isolates. Six isolates might have utilized other efflux pumps or antimicrobial resistance mechanisms. These analyses demonstrated a highly diverse population and the presence of complex resistance mechanisms in urinary isolates of K. pneumoniae.

scholarly journals Exogenous Isolation of Antibiotic Resistance Plasmids from Piggery Manure Slurries Reveals a High Prevalence and Diversity of IncQ-Like Plasmids

2000 ◽  
Vol 66 (11) ◽  
pp. 4854-4862 ◽  
Author(s):  
Kornelia Smalla ◽  
Holger Heuer ◽  
Antje Götz ◽  
Dagmar Niemeyer ◽  
Ellen Krögerrecklenfort ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Ralstonia Eutropha ◽  
Blot Hybridization ◽  
Restriction Pattern ◽  
Pcr Analysis ◽  
Dot Blot ◽  
High Prevalence ◽  
Resistance Plasmids ◽  
Incq Plasmids ◽  
K 12

ABSTRACT Antibiotic resistance plasmids were exogenously isolated in biparental matings with piggery manure bacteria as plasmid donors inEscherichia coli CV601 and Pseudomonas putidaUWC1 recipients. Surprisingly, IncQ-like plasmids were detected by dot blot hybridization with an IncQ oriV probe in severalP. putida UWC1 transconjugants. The capture of IncQ-like plasmids in biparental matings indicates not only their high prevalence in manure slurries but also the presence of efficiently mobilizing plasmids. In order to elucidate unusual hybridization data (weak or no hybridization with IncQ repB or IncQ oriTprobes) four IncQ-like plasmids (pIE1107, pIE1115, pIE1120, and pIE1130), each representing a different EcoRV restriction pattern, were selected for a more thorough plasmid characterization after transfer into E. coli K-12 strain DH5α by transformation. The characterization of the IncQ-like plasmids revealed an astonishingly high diversity with regard to phenotypic and genotypic properties. Four different multiple antibiotic resistance patterns were found to be conferred by the IncQ-like plasmids. The plasmids could be mobilized by the RP4 derivative pTH10 into Acinetobactersp., Ralstonia eutropha, Agrobacterium tumefaciens, and P. putida, but they showed diverse patterns of stability under nonselective growth conditions in different host backgrounds. Incompatibility testing and PCR analysis clearly revealed at least two different types of IncQ-like plasmids. PCR amplification of total DNA extracted directly from different manure samples and other environments indicated the prevalence of both types of IncQ plasmids in manure, sewage, and farm soil. These findings suggest that IncQ plasmids play an important role in disseminating antibiotic resistance genes.

scholarly journals Spo0A Suppresses sin Locus Expression in Clostridioides difficile

mSphere ◽  
2020 ◽  
Vol 5 (6) ◽  
Author(s):  
Babita Adhikari Dhungel ◽  
Revathi Govind
Keyword(s):  
Diarrheal Disease ◽  
Toxin Production ◽  
The United States ◽  
Upstream Region ◽  
Pcr Analysis ◽  
Bacterial Cells ◽  
Master Regulator ◽  
Content Type ◽  
Clostridioides Difficile

ABSTRACT Clostridioides difficile is the leading cause of nosocomial infection and is the causative agent of antibiotic-associated diarrhea. The severity of the disease is directly associated with toxin production, and spores are responsible for the transmission and persistence of the organism. Previously, we characterized sin locus regulators SinR and SinR′ (we renamed it SinI), where SinR is the regulator of toxin production and sporulation. The SinI regulator acts as its antagonist. In Bacillus subtilis, Spo0A, the master regulator of sporulation, controls SinR by regulating the expression of its antagonist, sinI. However, the role of Spo0A in the expression of sinR and sinI in C. difficile had not yet been reported. In this study, we tested spo0A mutants in three different C. difficile strains, R20291, UK1, and JIR8094, to understand the role of Spo0A in sin locus expression. Western blot analysis revealed that spo0A mutants had increased SinR levels. Quantitative reverse transcription-PCR (qRT-PCR) analysis of its expression further supported these data. By carrying out genetic and biochemical assays, we show that Spo0A can bind to the upstream region of this locus to regulates its expression. This study provides vital information that Spo0A regulates the sin locus, which controls critical pathogenic traits such as sporulation, toxin production, and motility in C. difficile. IMPORTANCE Clostridioides difficile is the leading cause of antibiotic-associated diarrheal disease in the United States. During infection, C. difficile spores germinate, and the vegetative bacterial cells produce toxins that damage host tissue. In C. difficile, the sin locus is known to regulate both sporulation and toxin production. In this study, we show that Spo0A, the master regulator of sporulation, controls sin locus expression. Results from our study suggest that Spo0A directly regulates the expression of this locus by binding to its upstream DNA region. This observation adds new detail to the gene regulatory network that connects sporulation and toxin production in this pathogen.

scholarly journals Stoichiometry and Turnover of the Bacterial Flagellar Switch Protein FliN

mBio ◽  
2014 ◽  
Vol 5 (4) ◽  
Author(s):  
Nicolas J. Delalez ◽  
Richard M. Berry ◽  
Judith P. Armitage
Keyword(s):  
Copy Number ◽  
Motor Proteins ◽  
Fluorescent Protein ◽  
Protein Complexes ◽  
Content Type ◽  
Rotation Direction ◽  
Cell Measurement ◽  
Biological Complexes ◽  
And Function

ABSTRACTSome proteins in biological complexes exchange with pools of free proteins while the complex is functioning. Evidence is emerging that protein exchange can be part of an adaptive mechanism. The bacterial flagellar motor is one of the most complex biological machines and is an ideal model system to study protein dynamics in large multimeric complexes. Recent studies showed that the copy number of FliM in the switch complex and the fraction of FliM that exchanges vary with the direction of flagellar rotation. Here, we investigated the stoichiometry and turnover of another switch complex component, FliN, labeled with the fluorescent protein CyPet, inEscherichia coli. Our results confirm that,in vivo, FliM and FliN form a complex with stoichiometry of 1:4 and function as a unit. We estimated that wild-type motors contained 120 ± 26 FliN molecules. Motors that rotated only clockwise (CW) or counterclockwise (CCW) contained 114 ± 17 and 144 ± 26 FliN molecules, respectively. The ratio of CCW-to-CW FliN copy numbers was 1.26, very close to that of 1.29 reported previously for FliM. We also measured the exchange of FliN molecules, which had a time scale and dependence upon rotation direction similar to those of FliM, consistent with an exchange of FliM-FliN as a unit. Our work confirms the highly dynamic nature of multimeric protein complexes and indicates that, under physiological conditions, these machines might not be the stable, complete structures suggested by averaged fixed methodologies but, rather, incomplete rings that can respond and adapt to changing environments.IMPORTANCEThe flagellum is one of the most complex structures in a bacterial cell, with the core motor proteins conserved across species. Evidence is now emerging that turnover of some of these motor proteins depends on motor activity, suggesting that turnover is important for function. The switch complex transmits the chemosensory signal to the rotor, and we show, by using single-cell measurement, that both the copy number and the fraction of exchanging molecules vary with the rotational bias of the rotor. When the motor is locked in counterclockwise rotation, the copy number is similar to that determined by averaged, fixed methodologies, but when locked in a clockwise direction, the number is much lower, suggesting that that the switch complex ring is incomplete. Our results suggest that motor remodeling is an important component in tuning responses and adaptation at the motor.

Neurodevelopment in the first three years: implications for child development, professional practice and policy

2014 ◽  
Vol 9 (2) ◽  
pp. 154-164 ◽  
Author(s):  
Danya Glaser
Keyword(s):  
Brain Development ◽  
Brain Activity ◽  
Brain Maturation ◽  
Policy And Practice ◽  
Content Type ◽  
Key Issues ◽  
And Function ◽  
The Relationship ◽  
The Brain ◽  
Brain Connections

Purpose – The purpose of this paper is to outline brain structure and development, the relationship between environment and brain development and implications for practice. Design/methodology/approach – The paper is based on a selected review of the literature and clinical experience. Findings – While genetics determine the sequence of brain maturation, the nature of brain development and functioning is determined by the young child's caregiving environment, to which the developing brain constantly adapts. The absence of input during sensitive periods may lead to later reduced functioning. There is an undoubted immediate equivalence between every mind function – emotion, cognition, behaviour and brain activity, although the precise location of this in the brain is only very partially determinable, since brain connections and function are extremely complex. Originality/value – This paper provides an overview of key issues in neurodevelopment relating to the development of young children, and implications for policy and practice.

scholarly journals Molecular Diversity of the Syndinean Genus Euduboscquella Based on Single-Cell PCR Analysis

2011 ◽  
Vol 78 (2) ◽  
pp. 334-345 ◽  
Author(s):  
Tsvetan R. Bachvaroff ◽  
Sunju Kim ◽  
Laure Guillou ◽  
Charles F. Delwiche ◽  
D. Wayne Coats
Keyword(s):  
Large Subunit ◽  
Small Subunit ◽  
Base Change ◽  
Pcr Analysis ◽  
Ssu Rrna ◽  
Compensatory Base Change ◽  
Content Type ◽  
Single Cell Pcr ◽  
Group I ◽  
Lsu Rrna

ABSTRACTThe genusEuduboscquellais one of a few described genera within the syndinean dinoflagellates, an enigmatic lineage with abundant diversity in marine environmental clone libraries based on small subunit (SSU) rRNA. The region composed of the SSU through to the partial large subunit (LSU) rRNA was determined from 40 individual tintinnid ciliate loricae infected withEuduboscquellasampled from eight surface water sites in the Northern Hemisphere, producing seven distinct SSU sequences. The corresponding host SSU rRNA region was also amplified from eight host species. The SSU tree ofEuduboscquellaand syndinean group I sequences from environmental clones had seven well-supported clades and one poorly supported clade across data sets from 57 to 692 total sequences. The genusEuduboscquellaconsistently formed a supported monophyletic clade within a single subclade of group I sequences. For most parasites with identical SSU sequences, the more variable internal transcribed spacer (ITS) to LSU rRNA regions were polymorphic at 3 to 10 sites. However, inE. cachonithere was variation between ITS to LSU copies at up to 20 sites within an individual, while in a parasite ofTintinnopsisspp., variation between different individuals ranged up to 19 polymorphic sites. However, applying the compensatory base change model to the ITS2 sequences suggested no compensatory changes within or between individuals with the same SSU sequence, while one to four compensatory changes between individuals with similar but not identical SSU sequences were found. Comparisons between host and parasite phylogenies do not suggest a simple pattern of host or parasite specificity.

scholarly journals Pyrobaculum yellowstonensis Strain WP30 Respires on Elemental Sulfur and/or Arsenate in Circumneutral Sulfidic Geothermal Sediments of Yellowstone National Park

2015 ◽  
Vol 81 (17) ◽  
pp. 5907-5916 ◽  
Author(s):  
Z. J. Jay ◽  
J. P. Beam ◽  
A. Dohnalkova ◽  
R. Lohmayer ◽  
B. Bodle ◽  
...  
Keyword(s):  
Yellowstone National Park ◽  
National Park ◽  
Elemental Sulfur ◽  
De Novo ◽  
Hot Spring ◽  
Content Type ◽  
Physiological Attributes ◽  
And Function ◽  
Metagenome Sequence

ABSTRACTThermoproteales(phylumCrenarchaeota) populations are abundant in high-temperature (>70°C) environments of Yellowstone National Park (YNP) and are important in mediating the biogeochemical cycles of sulfur, arsenic, and carbon. The objectives of this study were to determine the specific physiological attributes of the isolatePyrobaculum yellowstonensisstrain WP30, which was obtained from an elemental sulfur sediment (Joseph's Coat Hot Spring [JCHS], 80°C, pH 6.1, 135 μM As) and relate this organism to geochemical processes occurringin situ. Strain WP30 is a chemoorganoheterotroph and requires elemental sulfur and/or arsenate as an electron acceptor. Growth in the presence of elemental sulfur and arsenate resulted in the formation of thioarsenates and polysulfides. The complete genome of this organism was sequenced (1.99 Mb, 58% G+C content), revealing numerous metabolic pathways for the degradation of carbohydrates, amino acids, and lipids. Multiple dimethyl sulfoxide-molybdopterin (DMSO-MPT) oxidoreductase genes, which are implicated in the reduction of sulfur and arsenic, were identified. Pathways for thede novosynthesis of nearly all required cofactors and metabolites were identified. The comparative genomics ofP. yellowstonensisand the assembled metagenome sequence from JCHS showed that this organism is highly related (∼95% average nucleotide sequence identity) toin situpopulations. The physiological attributes and metabolic capabilities ofP. yellowstonensisprovide an important foundation for developing an understanding of the distribution and function of these populations in YNP.

Reflections on team culture, structure and function of an intensive support service centred on positive behavioural support

2016 ◽  
Vol 21 (4) ◽  
pp. 203-211 ◽  
Author(s):  
Lawrence A. Patterson ◽  
Samuel Berry
Keyword(s):  
Social Services ◽  
Support Service ◽  
Structure And Function ◽  
Content Type ◽  
Case Examples ◽  
Behavioural Support ◽  
Team Culture ◽  
Shared Identity ◽  
The Right ◽  
And Function

Purpose The purpose of this paper is to explore experiences of team culture, structure and function of an intensive support service (ISS) within the context of the recent service guidance “Building the Right Support” (NHS England, Local Government Association and Association of Directors of Adult Social Services, 2015). Reflections on the Hampshire and Southampton ISS set up in 2010 are discussed with a view to informing a debate about frameworks for ISS services nationally. Design/methodology/approach A reflective piece, drawing on experience and case examples. Findings This paper describes that a key function of an ISS is making individuals safe and this is significantly assisted by using shared team formulation, which can enable information and perspectives to be shared between and within teams as rapidly as possible. Further, a case is made for recognising the importance of inter-disciplinary practice, as the Southampton and Hampshire ISS has removed the “old fashioned” demarcations that led to individuals seeing a “procession” of different professionals from different disciplines. This relates to team structure, but importantly is about a culture of holding a shared identity based on positive behavioural support values, rather than a traditional uni-disciplinary perspective. Practical implications ISS models are being proposed by NHS England and this paper suggests some important practical aspects. Originality/value Limited literature exists examining the team culture within ISSs, which contributes to desired outcomes for service users. This paper opens a debate about structural and functional aspects of service delivery in this service model.

Prisoner’s insomnia prevalence, insomnia associated factors and interventions with sleep as an outcome: a review and narrative analysis

2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Chris Griffiths ◽  
Farah Hina
Keyword(s):  
Narrative Analysis ◽  
Associated Factors ◽  
Behavioural Therapy ◽  
Post Traumatic Stress ◽  
Content Type ◽  
Cognitive Behavioural ◽  
High Prevalence ◽  
Randomised Control Trials ◽  
Post Traumatic ◽  
Anxiety Depression

Purpose Insomnia is highly prevalent in prisoners. The purpose of this paper is a review of research evidence on interventions with sleep as an outcome (2000 to 2020) and rates of insomnia prevalence and associated factors in prisons (2015 to 2020). Design/methodology/approach An internet-based search used Medline, PubMed, PsycINFO (EBSCOhost), Embase, Web of Science and Scopus. Seven interventions and eight sleep prevalence or sleep-associated factor papers were identified. Findings Intervention research was very limited and the quality of the research design was generally poor. Interventions such as cognitive behavioural therapy for insomnia (CBT-I), yoga and mindfulness can be beneficial in a prison setting. This review identified a high prevalence of insomnia in prisons across the world, which was supported by recent evidence. Factors associated with insomnia include anxiety, depression, post-traumatic stress disorder, personality disorder and pain. Research limitations/implications There is a need for appropriately powered randomised control trials of CBT-I in prisons and a need to use objective measures of sleep quality. Originality/value Due to a lack of an up-to-date review, this paper fulfils the need for a review of the evidence on interventions in prison settings with sleep as an outcome, rates of insomnia prevalence and associated factors in prisons.

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