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1.
Sci Total Environ ; 922: 171158, 2024 Apr 20.
Article in English | MEDLINE | ID: mdl-38387558

ABSTRACT

Soil porosity and its reciprocal bulk density are important environmental state variables that enable modelers to represent hydraulic function and carbon storage. Biotic effects and their 'dynamic' influence on such state variables remain largely unknown for larger scales and may result in important, yet poorly quantified environmental feedbacks. Existing representation of hydraulic function is often invariant to environmental change and may be poor in some systems, particularly non-arable soils. Here we assess predictors of total porosity across two comprehensive national topsoil (0-15 cm) data sets, covering the full range of soil organic matter (SOM) and habitats (n = 1385 & n = 2570), using generalized additive mixed models and machine learning. Novel aspects of this work include the testing of metrics on aggregate size and livestock density alongside a range of different particle size distribution metrics. We demonstrate that porosity trends in Great Britain are dominated by biotic metrics, soil carbon and land use. Incorporating these variables into porosity prediction improves performance, paving the way for new dynamic calculation of porosity using surrogate measures with remote sensing, which may help improve prediction in data sparse regions of the world. Moreover, dynamic calculation of porosity could support representation of feedbacks in environmental and Earth System Models. Representing the hydrological feedbacks from changes in structural porosity also requires data and models at appropriate spatial scales to capture conditions leading to near-saturated soil conditions. Classification. Environmental Sciences.

2.
Sci Rep ; 12(1): 7085, 2022 04 30.
Article in English | MEDLINE | ID: mdl-35490195

ABSTRACT

The thin layer of soil at the earth's surface supports life, storing water and nutrients for plant uptake. These processes occur in the soil pore space, often half the soil volume, but our understanding of how this volume responds to environmental change is poor. Convention, has been to predict soil porosity, or its reciprocal bulk density (BD), from soil texture using pedotransfer functions (PTFs). A texture based approach, invariant to environmental change, prevents feedback from land use or climate change to soil porosity. Moreover, PTFs are often limited to mineral soils with < 20% soil organic matter (SOM) content. Here, we develop an analytical model to predict soil porosity, or BD, as a function of SOM. We test it on two comprehensive, methodologically consistent, temperate national-scale topsoil data sets (0-15 cm) (Wales, n = 1385; Great Britain, n = 2570). The purpose of the approach is to generate an analytical function suitable for predicting soil porosity change with SOM content, while providing insight into the main grain-scale factors determining the porosity emergence. The newly developed function covering the entire SOM gradient allows for impacts of land use, management or climate change to feedback on soil porosity or bulk density through decadal dynamic changes in SOM.


Subject(s)
Plants , Soil , Minerals , Porosity , Water
3.
Sci Rep ; 12(1): 1379, 2022 01 26.
Article in English | MEDLINE | ID: mdl-35082379

ABSTRACT

Soil organic carbon (SOC) concentration is the fundamental indicator of soil health, underpinning food production and climate change mitigation. SOC storage is highly sensitive to several dynamic environmental drivers, with approximately one third of soils degraded and losing carbon worldwide. Digital soil mapping illuminates where hotspots of SOC storage occur and where losses to the atmosphere are most likely. Yet, attempts to map SOC often disagree. Here we compare national scale SOC concentration map products to reveal agreement of data in mineral soils, with progressively poorer agreement in organo-mineral and organic soils. Divergences in map predictions from each other and survey data widen in the high SOC content land types we stratified. Given the disparities are highest in carbon rich soils, efforts are required to reduce these uncertainties to increase confidence in mapping SOC storage and predicting where change may be important at national to global scales. Our map comparison results could be used to identify SOC risk where concentrations are high and should be conserved, and where uncertainty is high and further monitoring should be targeted. Reducing inter-map uncertainty will rely on addressing statistical limitations and including covariates that capture convergence of physical factors that produce high SOC contents.

4.
Faraday Discuss ; 210(0): 9-28, 2018 10 01.
Article in English | MEDLINE | ID: mdl-30264833

ABSTRACT

The development of nanoscale electrochemistry since the mid-1980s has been predominately coupled with steady-state voltammetric (i-E) methods. This research has been driven by the desire to understand the mechanisms of very fast electrochemical reactions, by electroanalytical measurements in small volumes and unusual media, including in vivo measurements, and by research on correlating electrocatalytic activity, e.g., O2 reduction reaction, with nanoparticle size and structure. Exploration of the behavior of nanoelectrochemical structures (nanoelectrodes, nanoparticles, nanogap cells, etc.) of a characteristic dimension λ using steady-state i-E methods generally relies on the well-known relationship, λ2 ∼ Dt, which relates diffusional lengths to time, t, through the coefficient, D. Decreasing λ, by performing measurements at a nanometric length scales, results in a decrease in the effective timescale of the measurement, and provides a direct means to probe the kinetics of steps associated with very rapid electrochemical reactions. For instance, steady-state voltammetry using a nanogap twin-electrode cell of characteristic width, λ ∼ 10 nm, allows investigations of events occurring at timescales on the order of ∼100 ns. Among many other advantages, decreasing λ also increases spatial resolution in electrochemical imaging, e.g., in scanning electrochemical microscopy, and allows probing of the electric double layer. This Introductory Lecture traces the evolution and driving forces behind the "λ2 ∼ Dt" steady-state approach to nanoscale electrochemistry, beginning in the late 1950s with the introduction of the rotating ring-disk electrode and twin-electrode thin-layer cells, and evolving to current-day investigations using nanoelectrodes, scanning nanocells for imaging, nanopores, and nanoparticles. The recent focus on so-called "single-entity" electrochemistry, in which individual and very short redox events are probed, is a significant departure from the steady-state approach, but provides new opportunities to probe reaction dynamics. The stochastic nature of very fast single-entity events challenges current electrochemical methods and modern electronics, as illustrated using recent experiments from the authors' laboratory.


Subject(s)
Electrochemical Techniques/instrumentation , Nanotechnology/instrumentation , DNA/chemistry , Diffusion , Equipment Design , Kinetics , Microelectrodes , Microscopy, Scanning Tunneling/instrumentation , Models, Molecular , Nanoparticles/chemistry , Nanopores/ultrastructure , Oxidation-Reduction , Stochastic Processes
5.
Eur J Clin Microbiol Infect Dis ; 36(10): 1787-1793, 2017 Oct.
Article in English | MEDLINE | ID: mdl-28474178

ABSTRACT

Staphylococcus aureus is one of the most common causes of skin and soft tissue infections in health-care and community settings, but transmission of S. aureus in community-based populations is incompletely understood. S. aureus carriage phenotypes (persistent, intermittent, and non-carriers) were determined for households from Starr County, TX. Nasal swabs were collected from a cohort of 901 residents and screened for the presence of S. aureus. Isolated strains were spa-typed and assigned to clonal complexes. Of the 901 participants there were 134 pairs, 28 trios, 11 quartets, 3 quintets and 1 septet residing in the same household. There was a significant increase in "ever" carriers (persistent and intermittent carriers combined) in these households over that expected based on population frequencies (p = 0.029). There were 42 ever carrier pairs of individuals with 21 concordant for clonal complex type whereas only 4.7 were expected to be so (p = 6.9E-11). These results demonstrated clear aggregation of S. aureus carriage and concordance for strain types within households. As antibiotic-resistant S. aureus strains increase in community settings, it is important to better understand risk factors for colonization, mechanisms of transmission, clonal complexes present, and the role of household concordance/transmission.


Subject(s)
Carrier State/epidemiology , Family Health , Genotype , Methicillin Resistance , Staphylococcal Infections/epidemiology , Staphylococcus aureus/classification , Staphylococcus aureus/drug effects , Adolescent , Adult , Aged , Aged, 80 and over , Carrier State/microbiology , Family Characteristics , Female , Humans , Male , Middle Aged , Molecular Epidemiology , Molecular Typing , Prospective Studies , Staphylococcal Infections/microbiology , Staphylococcus aureus/genetics , Staphylococcus aureus/isolation & purification , Texas/epidemiology , Young Adult
6.
Faraday Discuss ; 193: 293-312, 2016 12 12.
Article in English | MEDLINE | ID: mdl-27711841

ABSTRACT

Here we investigate the mechanistic aspects of Pt nanoparticle (NP) aggregation in solutions typically used for detecting NP/electrode impacts by electrocatalytic amplification (ECA). We previously proposed a general mechanism for Pt colloid destabilization that involved the participation of both the hydrazine redox probe and the pH buffer species as coagulants. Herein the Pt NP coagulation and aggregation mechanisms were further investigated with microscopic kinetic NP concentration monitoring and zeta potential measurements using nanoparticle tracking analysis (NTA), as well as open circuit potential experiments with a citrate-treated polycrystalline Pt surface to assess electrical double layer potential. After considering the combined results of these experiments we propose that the colloidal stability of citrate-capped platinum nanoparticles involves much more than the typical physicochemical interactions predicted by DLVO theory. A structure based on intermolecular H-bonding in the citrate capping layer is the most plausible explanation for the exceptional stability of large Pt NPs in high ionic strength buffers. Thus, the mechanism of Pt NP aggregation includes specific reactive contributions from hydrazine. The catalytic decomposition of hydrazine, in particular, is thought to occur to some extent at the citrate-coated Pt surface while the citrate remains adsorbed. Evolved gases such as ammonia and possible surface bound intermediates from Pt-catalyzed decomposition of hydrazine may disrupt the stability of the citrate layer, causing colloidal instability and thus promoting Pt NP coagulation. In the closing section, we demonstrate nanoparticle impact electroanalysis by ECA detection as a method to quantify Pt NP concentration with adequate time resolution for monitoring the kinetics of Pt NP coagulation.

7.
J Vet Pharmacol Ther ; 38(6): 575-80, 2015 Dec.
Article in English | MEDLINE | ID: mdl-25846945

ABSTRACT

The objective was to compare plasma lidocaine concentrations when a commercially available 5% lidocaine patch was placed on intact skin vs. an incision. Our hypothesis was that greater absorption of lidocaine would occur from the incision site compared to intact skin. Ten dogs were used in a crossover design. A patch was placed over an incision, and then after a washout period, a patch was placed over intact skin. Plasma lidocaine concentrations were measured at patch placement; 20, 40 and 60 min; and 2, 4, 6, 12, 24, 36, 48, 72 and 96 h after patch placement. After patch removal, the skin was graded using a subjective skin reaction system. No dogs required rescue analgesia, and no toxicity or skin reaction was noted. Mean ± SD AUC and CMAX were 3054.29 ± 1095.93 ng·h/mL and 54.1 ± 15.84 ng/mL in the Incision Group, and 2269.9 ± 1037.08 ng·h/mL and 44.5 ± 16.34 ng/mL in the No-Incision Group, respectively. The AUC was significantly higher in the Incision Group. The results of the study demonstrate that the actual body exposure to lidocaine was significantly higher when an incision was present compared to intact skin. No adverse effects were observed from either treatment. Efficacy was not evaluated.


Subject(s)
Anesthetics, Local/administration & dosage , Dogs/surgery , Lidocaine/administration & dosage , Skin/metabolism , Transdermal Patch , Anesthetics, Local/blood , Anesthetics, Local/pharmacokinetics , Animals , Dogs/blood , Female , Lidocaine/blood , Lidocaine/pharmacokinetics , Male
8.
Clin Microbiol Infect ; 21(5): 444-50, 2015 May.
Article in English | MEDLINE | ID: mdl-25708549

ABSTRACT

Typing of healthcare-associated methicillin-resistant Staphylococcus aureus (MRSA) from Australia in the 1970s revealed a novel clone, ST2249-MRSA-III (CC45), present from 1973 to 1979. This clone was present before the Australian epidemic caused by the recombinant clone, ST239-MRSA-III. This study aimed to characterize the genome of ST2249-MRSA-III to establish its relationship to other MRSA clones. DNA microarray analysis was conducted and a draft genome sequence of ST2249 was obtained. The recombinant structure of the ST2249 genome was revealed by comparisons to publicly available ST239 and ST45 genomes. Microarray analysis of genomic DNA of 13 ST2249 isolates showed gross similarities with the ST239 chromosome in a segment around the origin of replication and with ST45 for the remainder of the chromosome. Recombination breakpoints were precisely determined by the changing pattern of nucleotide polymorphisms in the genome sequence of ST2249 isolate SK1585 compared with ST239 and ST45. One breakpoint was identified to the right of oriC, between sites 1014 and 1065 of the gene D484_00045. Another was identified to the left of oriC, between sites 1185 and 1248 of D484_01632. These results indicate that ST2249 inherited approximately 35.3% of its chromosome from an ST239-like parent and 64.7% from an ST45-like parent. ST2249-MRSA-III resulted from a major recombination between parents that resemble ST239 and ST45. Although only limited Australian archival material is available, the oldest extant isolate of ST2249 predates the oldest Australian isolate of ST239 by 3 years. It is therefore plausible that these two recombinant clones were introduced into Australia separately.


Subject(s)
Cross Infection/epidemiology , Cross Infection/microbiology , Genotype , Methicillin-Resistant Staphylococcus aureus/classification , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Recombination, Genetic , Staphylococcal Infections/epidemiology , Australia/epidemiology , Chromosomes, Bacterial , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Evolution, Molecular , Genome, Bacterial , Humans , Methicillin-Resistant Staphylococcus aureus/genetics , Microarray Analysis , Sequence Analysis, DNA , Staphylococcal Infections/microbiology
9.
Epidemiol Infect ; 143(11): 2430-9, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25434806

ABSTRACT

The aims of this study were to identify Staphylococcus aureus nasal colonization prevalence, behavioural risk factors, and to determine staphylococcal protein A (spa) types in community-based injection drug users (IDUs). Nasal swabs were collected and methicillin susceptibility testing and spa/SCCmec typing were performed on S. aureus isolates. Generalized estimating equations were used to report adjusted odds ratios and 95% confidence intervals. Of the 440 participants, 24·1% were colonized and 5·7% had methicillin-resistant S. aureus (MRSA). Colonization was associated with age, employment/marital status, and the presence of scabs but not with sexually transmitted disease co-infection, HIV status, antibiotic use, hospitalization, or drug treatment programme participation. The USA300 MRSA clone spa types were most common, but 15/49 spa types were new to one of the international databases. Community-based IDUs appear to have different risk factors compared to IDUs from clinical studies. In addition, the number of newly identified spa types indicates a diverse, understudied population.


Subject(s)
Carrier State/epidemiology , DNA, Bacterial/genetics , Nasal Mucosa/microbiology , Staphylococcal Infections/epidemiology , Staphylococcus aureus/isolation & purification , Substance Abuse, Intravenous/epidemiology , Adult , Age Factors , Anti-Bacterial Agents , Asymptomatic Infections/epidemiology , Bacterial Proteins/genetics , Carrier State/microbiology , Coinfection/epidemiology , Cross-Sectional Studies , Employment/statistics & numerical data , Female , HIV Infections/epidemiology , Ill-Housed Persons , Hospitalization/statistics & numerical data , Humans , Male , Marital Status/statistics & numerical data , Methicillin-Resistant Staphylococcus aureus/genetics , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Middle Aged , Penicillin-Binding Proteins , Prevalence , Risk Factors , Sexually Transmitted Diseases/epidemiology , Staphylococcal Infections/microbiology , Staphylococcal Protein A/genetics , Staphylococcus aureus/genetics , United States/epidemiology
10.
Epidemiol Infect ; 140(10): 1809-12, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22475374

ABSTRACT

A cluster of methicillin-resistant Staphylococcus aureus (MRSA) breast abscesses in women who had given birth at a hospital in Mumbai, India was investigated retrospectively. Nineteen of 20 cases were caused by a single clone: pvl-positive, spa type 648 (Ridom t852), ccrB:dru subtype 3:0, ST22-MRSA-IV. Despite the presence of pvl and SCCmec type IV, which are common genetic markers in community-associated MRSA, this outbreak was caused by a healthcare-associated, community-onset MRSA that was common in the hospital environment. Thus, infection control practices may have an important role in limiting the spread of this virulent clone.


Subject(s)
Abscess/epidemiology , Disease Outbreaks , Mastitis/epidemiology , Methicillin-Resistant Staphylococcus aureus/classification , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Staphylococcal Infections/epidemiology , Abscess/microbiology , Cluster Analysis , Female , Humans , India/epidemiology , Mastitis/microbiology , Methicillin-Resistant Staphylococcus aureus/genetics , Molecular Typing , Postpartum Period , Pregnancy , Retrospective Studies , Staphylococcal Infections/microbiology , Virulence Factors/genetics
11.
Eur J Clin Microbiol Infect Dis ; 31(10): 2559-66, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22450741

ABSTRACT

The molecular fingerprinting of a collection of 94 Staphylococcus aureus isolates from patients with osteomyelitis in Argentina was performed. Twenty-three SmaI pulsed-field gel electrophoresis (PFGE) types and 37 spa types were identified. The isolates were assigned to 23 sequence types (STs). The proportion of methicillin-resistant S. aureus (MRSA) isolates was significantly higher among cap5 S. aureus (35/61) compared with cap8 S. aureus (8/33) isolates (p = 0.0025). Twenty-four of the 94 isolates carried the lukS-PV/lukF-PV genes, which were significantly associated to cap5 [(23/38) compared with cap8 S. aureus isolates (1/32) (p = 0.0001)]. Forty of the 94 isolates carried genes of the egc locus (seg/sei). The distribution of seg/sei genes among isolates was related to certain clones. Isolates of the four agr types were found in the S. aureus collection. Whereas agr I isolates were evenly distributed among cap5 and cap8 S. aureus isolates (32/61 and 14/33, respectively), the agr II group was composed of 29 cap5 S. aureus isolates and agr III was composed of 16 cap8 S. aureus isolates. Two clones originally associated to animals (ST 188, 7 isolates and ST 1796, 5 isolates) were associated with chronic osteomyelitis and lack of capsular polysaccharide (CP) production. Loss of CP production remains the single factor among those investigated that is associated with chronic osteomyelitis.


Subject(s)
Bacterial Proteins/genetics , Osteomyelitis/microbiology , Polysaccharides, Bacterial/genetics , Staphylococcus aureus/isolation & purification , Virulence Factors/genetics , Argentina/epidemiology , Bacterial Typing Techniques , Electrophoresis, Gel, Pulsed-Field , Enterotoxins/genetics , Genes, Bacterial , Genetic Loci , Humans , Penicillin-Binding Proteins , Prevalence , Staphylococcal Infections/epidemiology , Staphylococcal Infections/microbiology , Staphylococcus aureus/classification , Staphylococcus aureus/genetics , Superantigens/genetics , Trans-Activators/genetics
12.
Sci Total Environ ; 434: 62-70, 2012 Sep 15.
Article in English | MEDLINE | ID: mdl-22245213

ABSTRACT

The amount of plant-available nitrogen (N) in soil is an important indicator of eutrophication of semi-natural habitats, but previous studies have shown contrasting effects of N deposition on mineralisable N in different habitats. The stock of readily mineralisable N (N(rm)) was measured in 665 locations across Britain from a range of intensively and extensively managed habitats, allowing N availability to be studied in relation to soil and vegetation type, and also to variation in climate and in reactive N deposition from the atmosphere. Mineralisable N contents were correlated with deposition in extensively managed habitats but not in intensively managed habitats. The following statements apply only to extensively managed habitats. All habitats showed a similar increase in N(rm) with N deposition. However, soil characteristics affected the relationship, and soil carbon content in particular was a major control on mineralisation. The N(rm) stock increased more with N deposition in organic than in mineral soils. The nitrate proportion of N(rm) also increased with N deposition but, conversely, this increase was greater in mineral than in organic soils. The measurements could be used as indicators of eutrophication, e.g. deposition rates of over 20 kg N ha(-1) y(-1) are associated with nitrate proportions of >41% in a mineral soil (2% carbon), and with N(rm) stocks of over 4.8 kg N ha(-1) in an organic soil (55% carbon). Both N(rm) and nitrate proportion increased with mean annual temperature of the sampling location, despite consistent incubation temperature, suggesting that increasing temperatures are likely to increase the eutrophying effects of N pollution on semi-natural ecosystems.


Subject(s)
Climate , Ecosystem , Nitrogen/chemistry , Soil , Nitrogen Fixation
13.
J Bone Joint Surg Br ; 92(10): 1397-402, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20884978

ABSTRACT

We describe the clinical outcome of a technique of surgical augmentation of chronic massive tears of the rotator cuff using a polyester ligament (Dacron) in 21 symptomatic patients (14 men, seven women) with a mean age of 66.5 years (55.0 to 85.0). All patients had MRI and arthroscopic evidence of chronic massive tears. The clinical outcome was assessed using the Constant and Murley and patient satisfaction scores at a mean follow-up of 36 months (30 to 46). The polyester ligament (500 mm × 10 mm) was passed into the joint via the portal of Neviaser, medial to the tear through healthy cuff. The two ends of the ligament holding the cuff were passed through tunnels made in the proximal humerus at the footprint of the insertion of the cuff. The ligament was tied with a triple knot over the humeral cortex. All the patients remained free from pain (p < 0.001) with improvement in function (p < 0.001) and range of movement (p < 0.001). The mean pre-operative and post-operative Constant scores were 46.7 (39.0 to 61.0) and 85.4 (52.0 to 96.0), respectively (p < 0.001). The mean patient satisfaction score was 90%. There were two failures, one due to a ruptured ligament after one year and the other due to deep-seated infection. The MR scan at the final follow-up confirmed intact and thickened bands in 15 of 17 patients. This technique of augmentation gives consistent relief from pain with improved shoulder movement in patients with symptomatic massive tears of the rotator cuff.


Subject(s)
Ligaments, Articular/surgery , Prosthesis Implantation/methods , Rotator Cuff Injuries , Rotator Cuff/surgery , Aged , Aged, 80 and over , Female , Humans , Magnetic Resonance Imaging , Male , Middle Aged , Pain Measurement , Patient Satisfaction , Polyethylene Terephthalates , Prospective Studies , Prostheses and Implants , Range of Motion, Articular , Recovery of Function , Shoulder Joint/pathology , Shoulder Joint/physiopathology , Shoulder Pain/etiology , Shoulder Pain/surgery , Treatment Outcome
14.
Int J Obstet Anesth ; 18(4): 408-12, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19733476

ABSTRACT

There have been several reports of resistance to local anaesthetic agents in women with Ehlers-Danlos syndrome, hypermobility type, also known as Ehlers-Danlos syndrome Type III. General anaesthesia with rapid sequence induction was performed for caesarean section due to prolonged second stage of labour, but intubation proved to be difficult. We propose that intubation difficulty probably arose from collapse of fibro-elastic tissues and adjoining C-shaped cartilages of the trachea with appropriately applied cricoid pressure. We found no other case reports of difficult intubation in patients with Ehlers-Danlos syndrome, hypermobility type. There are reports of cervical spine instability and temporomandibular joint dysfunction in patients with this syndrome suggesting a potential for difficult airway management. Additional anaesthetic problems associated with Ehlers-Danlos syndrome involve patient positioning and vascular access.


Subject(s)
Anesthesia, Inhalation , Anesthesia, Obstetrical , Ehlers-Danlos Syndrome/complications , Intubation, Intratracheal , Adult , Cricoid Cartilage , Ehlers-Danlos Syndrome/therapy , Female , Humans , Infant, Newborn , Pregnancy , Pressure , Transcutaneous Electric Nerve Stimulation
15.
Gene Ther ; 12(4): 347-57, 2005 Feb.
Article in English | MEDLINE | ID: mdl-15538392

ABSTRACT

It is well established that mutations in specific structural elements of the motor protein myosin are directly linked to debilitating diseases involving malfunctioning striated muscle cells. A potential way to study the relationship between myosin structure and function is to express exogenous myosin in vivo and determine contractile properties of the transgenic muscle cells. However, in vivo expression of functional levels of contractile proteins using transient transgenesis in skeletal muscle has not been demonstrated. Presently, we used in vivo gene transfer to express high levels of full-length myosin light chain (MLC) in skeletal muscle fibers of Rana pipiens. Anterior tibialis (AT) muscles were injected with cardiotoxin to cause degeneration and then injected at various stages of regeneration with plasmid expression vectors encoding full-length MLC1(f). In fibers from the most robustly transfected muscles 3 weeks after plasmid injections, trans-MLC1(f) expression averaged 22-43% of the endogenous MLC1(f). Trans-MLC1(f) expression was the same whether a small epitope tag was placed on the C- or N-terminus and was highly variable along individual fibers. Confocal microscopy of skinned fibers showed correct sarcomeric incorporation of trans-MLC1(f). The expression profile of myosin heavy chain isoforms 21 days after transfection was similar to normal AT muscle. These data demonstrate the feasibility of using in vivo gene transfer to probe the structural basis of contractile protein function in skeletal muscle. Based on these promising results, we discuss how further improvements in the level and consistency of myosin transgene expression may be achieved in future studies, and the therapeutic potential of plasmid gene transfer in regenerating muscle.


Subject(s)
Genetic Therapy/methods , Muscle, Skeletal/metabolism , Muscular Dystrophies/therapy , Myosin Light Chains/genetics , Regeneration , Animals , Blotting, Western/methods , Gene Expression , Immunohistochemistry/methods , Male , Microscopy, Confocal , Plasmids , Rana pipiens , Transgenes
17.
J Clin Microbiol ; 42(7): 3185-90, 2004 Jul.
Article in English | MEDLINE | ID: mdl-15243080

ABSTRACT

Community methicillin-resistant Staphylococcus aureus (CMRSA) strains are being isolated with increasing frequency around the world. In Western Australia CMRSA are endemic in geographically remote communities and have been found to belong to five different contour-clamped homogeneous electric field (CHEF) electrophoretic patterns. Representatives of each of these CHEF patterns have been compared to CMRSA representative of CHEF patterns from other Australian states and New Zealand. With one exception, all of the isolates were nonmultiresistant and were not resistant to many antimicrobial agents other than the beta-lactams. With one exception, which is not believed to be a CMRSA, all of the isolates harbored a beta-lactamase plasmid. Erythromycin resistance was associated with a 2-kb plasmid. One of the beta-lactamase plasmids was found to be able to acquire additional resistance determinants to become a multiple resistance plasmid. There were 10 multilocus sequence types belonging to eight distantly related clonal complexes of S. aureus. One new sequence type was found. Although most of the CMRSA harbored the type IVa SCCmec, a type IV structural variant was found and two new SCCmec types were identified. Protein A gene (spa) typing revealed two new spa types and, with two exceptions, corresponded to multilocus sequence typing. In contrast to other reports on CMRSA, most of the CMRSA strains studied here did not contain the Panton-Valentine leukocidin genes. The results also demonstrate that nonmultiresistant hospital strains such as UK EMRSA-15 may be able to circulate in the community and could be mistaken for CMRSA based on their resistance profiles.


Subject(s)
Methicillin Resistance , Staphylococcus aureus/classification , Bacterial Typing Techniques , Drug Resistance, Multiple, Bacterial , Microbial Sensitivity Tests , Plasmids , Polymorphism, Restriction Fragment Length , Staphylococcus aureus/drug effects , Staphylococcus aureus/genetics
18.
J Invest Dermatol ; 123(1): 62-6, 2004 Jul.
Article in English | MEDLINE | ID: mdl-15191543

ABSTRACT

Atopic dermatitis is a disease with an impaired skin barrier that affects 15%-20% of children. In the normal epidermis, the stratum corneum chymotryptic enzyme (SCCE) thought to play a central role in desquamation by cleaving proteins of the stratum corneum (e.g., corneodesmosin and plakoglobin). Genetic variations within the SCCE gene could be associated with dysregulation of SCCE activity leading to an abnormal skin barrier. We screened the SCCE gene for variations and performed a case-control study on 103 atopic dermatitis patients and 261 matched controls. 16 synonymous single nucleotide polymorphisms (SNPs) have been identified and a 4 bp (AACC) insertion has been found in the 3'UTR. We performed an association study of the SCCE AACC insertion in the 3'UTR, and found a significant trend between the AACC allele with the two insertions and disease in the overall data set [odds ratio (OR)=2.31; p=0.0007]. The AACC insertion in the SCCE gene may result in a change to SCCE activity within the skin barrier. These findings suggest that SCCE could have an important role in the development of atopic dermatitis.


Subject(s)
Dermatitis, Atopic/genetics , Polymorphism, Single Nucleotide , Serine Endopeptidases/genetics , 3' Untranslated Regions/genetics , Case-Control Studies , Exons/genetics , Genotype , Humans , Introns/genetics , Kallikreins
19.
Clin Microbiol Infect ; 10(2): 92-7, 2004 Feb.
Article in English | MEDLINE | ID: mdl-14759234

ABSTRACT

The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in many countries is increasing and, in hospitals in some areas, more than half of all S. aureus disease isolates are MRSA. MRSA strains are becoming increasingly multiresistant, and have recently developed resistance to vancomycin, used successfully to treat MRSA for more than 30 years. This review summarises recent studies that have elucidated the evolutionary history of MRSA. The first MRSA isolate evolved from a sensitive, epidemic strain prevalent in Europe, and its progeny-the first MRSA clone-quickly spread to other continents. Analyses of epidemic MRSA isolates from hospitals in different countries by molecular methods, including multilocus sequence typing (MLST) and DNA microarray analysis, reveal that MRSA strains have evolved separately within five distinct epidemic, sensitive lineages. However, resistance has been transferred to S. aureus on many more than five occasions, as some lineages have acquired different structural types of the element carrying the methicillin resistance gene. The emergence of MRSA as a community pathogen has been noted in several countries, and MLST and SCCmec typing have been used to demonstrate that community-acquired MRSA strains are typically related only distantly to hospital MRSA strains, and thus represent novel acquisitions of SCCmec.


Subject(s)
Bacterial Proteins/genetics , Evolution, Molecular , Methicillin Resistance/genetics , Sequence Analysis, DNA , Staphylococcus aureus/classification , Staphylococcus aureus/drug effects , Bacterial Typing Techniques , Humans , Staphylococcal Infections/epidemiology , Staphylococcal Infections/microbiology , Staphylococcus aureus/genetics
20.
Biochem Soc Trans ; 31(Pt 3): 548-52, 2003 Jun.
Article in English | MEDLINE | ID: mdl-12773154

ABSTRACT

The background and current context of work on the shikimate-pathway enzymes as potential targets for anti-bacterial, anti-fungal and anti-parasitic drugs is reviewed. Recent work on the third enzyme of the pathway, dehydroquinase, which occurs in two structurally and mechanistically distinct forms, is used to illustrate the present state of studies into rational drug design.


Subject(s)
Enzyme Inhibitors/chemical synthesis , Hydro-Lyases/metabolism , Shikimic Acid/metabolism , Bacteria/drug effects , Bacteria/enzymology , Drug Design , Enzyme Inhibitors/pharmacology , Hydro-Lyases/antagonists & inhibitors , Hydro-Lyases/chemistry , Plants/enzymology
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