A one-hour universal protocol for mouse genotyping

Introduction Transgenic animals are widely used for research and for most of them, genotyping is unavoidable. Published protocols may be powerful but may also present disadvantages such as their cost or the requirement of additional steps/equipment. Moreover, if more than one strain must be genotyped, several protocols may need to be developed. Methods we adapted the existing amplification‐resistant mutation protocol to develop the 1‐hour universal genotyping protocol (1‐HUG), which allows the robust genotyping of genetically modified mice in 1 h from sample isolation to PCR gel running. Results This protocol allows the genotyping of different mouse models including mdx mouse, and FLExDUX4 and HSA‐MerCreMer alone or in combination. It can be applied to different types of genomic modifications and to sexing. Discussion The 1‐HUG protocol can be used routinely in any laboratory using mouse models for neuromuscular diseases

Inhalation therapy during acute asthma : the role of a combined steroid and beta-stimulant preparation

CITATION: Joubert, J. R., Burger, G. & Shephard, E. 1985. Inhalation therapy during acute asthma : the role of a combined steroid and beta-stimulant preparation. South African Medical Journal, 68:381-384.The original publication is available at http://www.samj.org.zaA compound consisting of a β-stimulant, salbutamol (100 μg/puff), and a steroid, beclomethasone diproprionate (50 μg/puff), was studied to test the hypothesis that the corticosteroid could enhance the bronchodilator proporties of the β-stimulant during chronic asthma and simulated acute attacks (antigen challenge). Conventional doses (200 μg and 100 μg of salbutamol and beclomethasone respectively) were compared using a schedule which included a second administration 1 hour later. The results obtained on the baseline bronchial responsiveness of chronic asthmatics and during the delayed asthmatic response (simulated acute asthma) were similar. The compound was as effective as salbutamol alone but not more so. A significantly greater bronchodilator response was recorded in all patients after the second administration of both the compound and salbutamol alone. The practical advantages of having one rather than two inhalers are evident, but the appropriate application of this compound agent, probably in a prophylactic role, must be defined.Publisher’s versio

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South Africa’s vital statistics are currently not suitable for monitoring progress towards injury and violence Sustainable Development Goal

Control of rheumatoid arthritis with conventional disease-modifying antirheumatic drugs in a tertiary hospital in central South Africa

Background. Rheumatoid arthritis (RA) is a highly prevalent disease with a significant impact on morbidity and life expectancy. There is a paucity of literature relating to the current state of disease control in South Africa, and none on Free State Province.Objectives. To evaluate control of RA with conventional disease-modifying antirheumatic drugs (DMARDs) in the rheumatology outpatient department of Universitas Academic Hospital, Bloemfontein, and to determine the relative impact of various factors contributing to the prevention of disease control.Methods. A cross-sectional study was undertaken over a period of 8 months from December 2016 to August 2017 in the rheumatology outpatient department of Universitas Academic Hospital. Data were collected by means of information sheets and questionnaires completed by attending doctors. The information pertained to disease activity, possible factors contributing to poor control, and relevant demographic data.Results. Information was collected from 169 patients, and data analysis was performed on 161. The results revealed that RA was controlled in 34 patients (21.1%). Of the 127 patients with uncontrolled disease, 61 (48.0%) reported dispensing issues relating to poor drug availability. Seventy-two (56.7%) of the patients with uncontrolled disease were deemed to be on insufficient treatment for their disease state, of whom 33 (45.8%) also reported concomitant dispensing issues. Other factors such as transport/access problems, administrative issues, adverse events and poor compliance/insight contributed to the inadequate control of RA to a minor extent. In terms of monthly dispensing, 159 patients (98.8%) reported receiving all their DMARDs during the first month. The vast majority of these drugs were dispensed from Universitas Hospital. This figure decreased to 119 patients (73.9%) who received all their DMARDs during the second month, mostly from district units. Only 55.3% (n=89) of the patients reported receiving all of their DMARDs from their down-referral units.Conclusions. Control of RA at this institution is suboptimal compared with national and international standards. The main contributors to poor control appear to be problems relating to dispensing of medication and inadequate escalation of therapy by doctors. Most of the concern with the dispensing of medication lies with the poor availability of DMARDs in peripheral unit pharmacies. These factors are remediable and should be attended to

Salt modulates the stability and lipid binding affinity of the adipocyte lipid-binding protein

Adipocyte lipid-binding protein (ALBP or aP2) is an intracellular fatty acid-binding protein that is found in adipocytes and macrophages and binds a large variety of intracellular lipids with high affinity. Although intracellular lipids are frequently charged, biochemical studies of lipid-binding proteins and their interactions often focus most heavily on the hydrophobic aspects of these proteins and their interactions. In this study, we have characterized the effects of KCl on the stability and lipid binding properties of ALBP. We find that added salt dramatically stabilizes ALBP, increasing its ΔG of unfolding by 3-5 kcal/mol. At 37 °C salt can more than double the stability of the protein. At the same time, salt inhibits the binding of the fluorescent lipid 1-anilino-naphthalene-8-sulfonate (ANS) to the protein and induces direct displacement of the lipid from the protein. Thermodynamic linkage analysis of the salt inhibition of ANS binding shows a nearly 1:1 reciprocal linkage: i.e. one ion is released from ALBP when ANS binds, and vice versa. Kinetic experiments show that salt reduces the rate of association between ANS and ALBP while simultaneously increasing the dissociation rate of ANS from the protein. We depict and discuss the thermodynamic linkages among stability, lipid binding, and salt effects for ALBP, including the use of these linkages to calculate the affinity of ANS for the denatured state of ALBP and its dependence on salt concentration. We also discuss the potential molecular origins and potential intracellular consequences of the demonstrated salt linkages to stability and lipid binding in ALBP

Exploring vibrational ladder climbing in vibronic coupling models: Toward experimental observation of a geometric phase signature of a conical intersection

Conical intersections (CIs) have been widely studied using spectroscopic techniques. However, CIs have mainly been identified by rapid internal conversion transitions that take place after the photoexcitation. Such identifications cannot distinguish various types of intersections as well as to separate the actual intersection from an avoided crossing. In this paper, we investigate how ultrafast IR laser pulses can be utilized to stimulate nuclear dynamics revealing geometric phase features associated with CIs. We consider two low-dimensional nonadiabatic models to obtain optimal two- and three-pulse laser sequences for stimulating nuclear dynamics necessary for the CI identification. Our results provide insights on designing non-linear spectroscopic schemes for subsequent probes of the nuclear wavepackets by ultrafast electron diffraction techniques to unambiguously detect CIs in molecules

An evaluation of the effect of an educational intervention for Australian social workers on competence in delivering brief cognitive behavioural strategies: A randomised controlled trial

Background: Broad community access to high quality evidence-based primary mental health care is an ongoing challenge around the world. In Australia one approach has been to broaden access to care by funding psychologists and other allied health care professionals to deliver brief psychological treatments to general practitioners' patients. To date, there has been a scarcity of studies assessing the efficacy of social worker delivered psychological strategies. This study aims to build the evidence base by evaluating the impact of a brief educational intervention on social workers' competence in delivering cognitive behavioural strategies (strategies derived from cognitive behavioural therapy). Methods. A randomised controlled trial design was undertaken with baseline and one-week follow-up measurement of both objective and self-perceived competence. Simulated consultations with standardised depressed patients were recorded on videotape and objective competence was assessed by blinded reviewers using the Cognitive Therapy Scale. Questionnaires completed by participants were used to measure self-perceived competence. The training intervention was a 15 hour face-to-face course involving presentations, video example consultations, written materials and rehearsal of skills in pairs. Results. 40 Melbourne-based (Australia) social workers enrolled and were randomised and 9 of these withdrew from the study before the pre training simulated consultation. 30 of the remaining 31 social workers (97%) completed all phases of the intervention and evaluation protocol (16 from intervention and 14 from control group). The intervention group showed significantly greater improvements than the control group in objective competence (mean improvement of 14.2 (7.38-21.02) on the 66 point Cognitive Therapy Scale) and in subjective confidence (mean improvement of 1.28 (0.84-1.72) on a 5 point Likert scale). On average, the intervention group improved from below to above the base competency threshold on the Cognitive Therapy Scale whilst the control group remained below. Conclusions. Social workers can attain significant improvements in competency in delivering cognitive behavioural strategies from undertaking brief face to face training. This is relevant in the context of health reforms that involve social worker delivery of evidence based psychological care. Further research is required to assess how these improvements in competence translate into performance in practice and clinical outcomes for patients

A Deoxyribonucleic Acid Decoy Trapping DUX4 for the Treatment of Facioscapulohumeral Muscular Dystrophy

Facioscapulohumeral dystrophy (FSHD) is characterized by a loss of repressive epigenetic marks leading to the aberrant expression of the DUX4 transcription factor. In muscle, DUX4 acts as a poison protein though the induction of multiple downstream genes. So far, there is no therapeutic solution for FSHD. Because DUX4 is a transcription factor, we developed an original therapeutic approach, based on a DNA decoy trapping the DUX4 protein, preventing its binding to genomic DNA and thereby blocking the aberrant activation of DUX4’s transcriptional network. In vitro, transfection of a DUX4 decoy into FSHD myotubes reduced the expression of the DUX4 network genes. In vivo, both double-stand DNA DUX4 decoys and adeno-associated viruses (AAVs) carrying DUX4 binding sites reduced transcriptional activation of genes downstream of DUX4 in a DUX4-expressing mouse model. Our study demonstrates, both in vitro and in vivo, the feasibility of the decoy strategy and opens new avenues of research

Structure Functions of the Nucleon in a Statistical Model

Deep inelastic scattering is considered in a statistical model of the nucleon. This incorporates certain features which are absent in the standard parton model such as quantum statistical correlations which play a role in the propagation of particles when considering Feynman diagrams containing internal lines. The inclusion of the ${\cal O}(\alpha_{s})$ corrections in our numerical calculations allows a good fit to the data for $x\geq 0.25$. The fit corresponds to values of temperature and chemical potential of approximately $T=0.067$ GeV and $\mu=0.133$ GeV. The latter values of parameters, however, give rise, for all $x$, to a large value for $R=\sigma_{L}/\sigma_{T}$.Comment: 16 pages TeX, 11 figures available as Postscript files, University of Bielefeld preprint BI-TP 93/3