CRISPR-Cas9-based approaches for genetic analysis and epistatic interaction studies in .

Unlabelled: is an obligate intracellular bacterial pathogen that replicates to high numbers in an acidified lysosome-derived vacuole. Intracellular replication requires the Dot/Icm type IVB secretion system, which translocates over 100 different effector proteins into the host cell. Screens employing random transposon mutagenesis have identified several effectors that play an important role in intracellular replication; however, the difficulty in conducting directed mutagenesis has been a barrier to the systematic analysis of effector mutants and to the construction of double mutants to assess epistatic interactions between effectors.

Necromass responses to warming: A faster microbial turnover in favor of soil carbon stabilisation.

Microbial byproducts and residues (hereafter 'necromass') potentially play the most critical role in soil organic carbon (SOC) sequestration. However, little is known about the influence of climate warming on necromass accumulation in the agroecosystem and the underlying mechanisms associated with microbial life strategies. In order to address these knowledge gaps, we used amino sugars as biomarkers of microbial necromass, and investigated their variation through an 8-year trial in an agroecosystem with two warming levels (+1.

Developmental Transitions Coordinate Assembly of the Coxiella burnetii Dot/Icm Type IV Secretion System.

Coxiella burnetii is an obligate intracellular bacterial pathogen that has evolved a unique biphasic developmental cycle. The infectious form of C. burnetii is the dormant small cell variant (SCV), which transitions to a metabolically active large cell variant (LCV) that replicates inside the lysosome-derived host vacuole.

Coxiella burnetii encodes an LvgA-related protein important for intracellular replication.

Coxiella burnetii is a bacterial pathogen that replicates in a specialised lysosome-derived organelle called the Coxiella-containing vacuole (CCV). Establishment of the CCV requires the Dot/Icm type IVB secretion system. A previous transposon mutagenesis screen identified the gene cbu1754 as being important for the intracellular replication of C.

Outside-in Repair Technique for a Complete Radial Tear of the Lateral Meniscus.

The lateral meniscus is vital in dissipating the force in the lateral compartment of the knee. A complete radial tear of the meniscus can lead to extrusion, rendering it nonfunctional and resulting in deleterious arthritic changes to the lateral compartment. Arthroscopic repair of a complete radial tear of the lateral meniscus poses a challenge to orthopaedic surgeons.

Narcotics and Knee Arthroscopy: Trends in Use and Factors Associated With Prolonged Use and Postoperative Complications.

Purpose: To (1) examine trends in the prevalence of preoperative and prolonged postoperative narcotic use in patients undergoing knee arthroscopy, (2) characterize factors associated with prolonged narcotic use after knee arthroscopy, and (3) explore the association of preoperative and prolonged postoperative narcotic use with complications after knee arthroscopy.

Methods: The PearlDiver database was reviewed for patients who underwent knee arthroscopy from 2007 to 2015 with a minimum of 6 months' follow up. Patients with preoperative or prolonged postoperative narcotic use were identified and analyzed for trends.

Efficiency and Safety: The Best Time to Valve a Plaster Cast.

Background: The act of applying, univalving, and spreading a plaster cast to accommodate swelling is commonly performed; however, cast saws can cause thermal and/or abrasive injury to the patient. This study aims to identify the optimal time to valve a plaster cast so as to reduce the risk of cast-saw injury and increase spreading efficiency.

Methods: Plaster casts were applied to life-sized pediatric models and were univalved at set-times of 5, 8, 12, or 25 minutes.

DNA Delivery and Genomic Integration into Mammalian Target Cells through Type IV A and B Secretion Systems of Human Pathogens.

We explore the potential of bacterial secretion systems as tools for genomic modification of human cells. We previously showed that foreign DNA can be introduced into human cells through the Type IV A secretion system of the human pathogen . Moreover, the DNA is delivered covalently attached to the conjugative relaxase TrwC, which promotes its integration into the recipient genome.

Detection of the diuretic hydrochlorothiazide in a doping control urine sample as the result of a non-steroidal anti-inflammatory drug (NSAID) tablet contamination.

Hydrochlorothiazide (HCTZ, 6-chloro-3,4-dihydro-2H-1,2,4-benzothiadiazine-7-sulfonamide-1,1-dioxide) belongs to the class of diuretic agents that represent one of today's cornerstones of the treatment of hypertensive patients. In addition to its clinical relevance, HCTZ is prohibited in sports according to the regulations of the World Anti-Doping Agency (WADA) at all times and has frequently been detected in sports drug testing urine samples worldwide since its ban was introduced in 1988. Despite these facts, the adverse analytical finding concerning HCTZ in an in-competition routine doping control sample collected in December 2014 was further investigated, particularly motivated by the comparably low urinary concentration of the drug accounting for approximately 5ng/mL.

Allosteric activation of exopolysaccharide synthesis through cyclic di-GMP-stimulated protein-protein interaction.

In many bacterial pathogens, the second messenger c-di-GMP stimulates the production of an exopolysaccharide (EPS) matrix to shield bacteria from assaults of the immune system. How c-di-GMP induces EPS biogenesis is largely unknown. Here, we show that c-di-GMP allosterically activates the synthesis of poly-β-1,6-N-acetylglucosamine (poly-GlcNAc), a major extracellular matrix component of Escherichia coli biofilms.

Second messenger signalling governs Escherichia coli biofilm induction upon ribosomal stress.

Biofilms are communities of surface-attached, matrix-embedded microbial cells that can resist antimicrobial chemotherapy and contribute to persistent infections. Using an Escherichia coli biofilm model we found that exposure of bacteria to subinhibitory concentrations of ribosome-targeting antibiotics leads to strong biofilm induction. We present evidence that this effect is elicited by the ribosome in response to translational stress.

The effects of botulinum toxin A on muscle histology during distraction osteogenesis.

Distraction osteogenesis is a highly successful method of bone formation, yet muscle fibrosis and contractures can result in significant morbidity. In the current study, we investigate the efficacy of botulinum toxin A in preventing fibrosis and potentially increasing muscle development in distracted muscles. Fifteen New Zealand White rabbits underwent tibial distraction at 1.