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1.
PLoS One ; 13(4): e0193550, 2018.
Article in English | MEDLINE | ID: mdl-29621274

ABSTRACT

BACKGROUND: Fabry disease (FD) results from X-linked inheritance of a mutation in the GLA gene, encoding for alpha galactosidase A, and is characterized by heterogeneous clinical manifestations. Two phenotypes have been described "Classic" and "late onset" which cannot be predicted exclusively by genotype. The latter has been considered an attenuated form of the disease often affecting a single organ system commonly the heart. Recent studies have demonstrated that cardiac outcomes are similar in patients with classic and late onset mutations. In this study we investigate the relationship between clinical heterogeneity and plasma lyso-Gb3 in a large single centre cohort of N215S patients and compare this to patients with other mutations. METHODS: In this single-centre, retrospective, cross-sectional study we analysed a cohort of 251 FD patients: 84 N215S mutation (37 males) and 167 non-N215S mutations (58 males). The Mainz severity score index (MSSI) was used as an index of overall disease severity. Cardiac function and morphology were assessed by electrocardiogram and echocardiogram. Left ventricular mass was calculated using the Devereux formula and the left ventricular mass index (LVMI) calculated to adjust for height (g/m2.7). The presence of white matter lesions was assessed by cerebral MRI or computed tomography (CT). GFR was measured by radio-isotope (chromium-EDTA) method and adjusted for patient height (ml/min/m2.7), and urinary protein quantification was undertaken by 24 hour urine collection. Plasma globotriaosylsphingosine (lyso-Gb3) was analysed prior to ERT in 84 patients. RESULTS: N215S patients showed later symptom onset (males: p< 0.0001, females: p<0.03), later development of left ventricular hypertrophy (LVH) (median survival without LVH: 41 (non-N215S) vs. 64 (N215S) years, p< 0.0001), later development of proteinuria (median survival without proteinuria 43 (non-N215S) vs 71 years (N215S), p< 0.0001), later occurrence of cerebrovascular events (stroke/ Transient Ischaemic Attacks (TIA); median survival without stroke: 74 years (non-N215S) vs. not reached (N215S), p< 0.02), later decline in renal function to GFR <60 ml/min/1.73m2 (median survival: 56 (non-N215S) vs. 72 (N215S) years, p< 0.01), and greater overall survival (median survival 81 (N215S) vs. 66 (non-N215S) years, p< 0.0006). Lyso-Gb3 was found to be less elevated in N215S compared to non-N215S male and female patients. However, the N215S population eventually reached an overall severity measured by MSSI comparable to the non-N215S without equivalent elevation of lyso-Gb3 (means: 6.7 vs. 74.3 nmol/L, p < 0.001). In addition, N215S patients showed strong correlations between lyso-Gb3 levels and LVMI, GFR, and MSSI. These associations became stronger when we investigated individuals' life time exposure to lyso-Gb3 (calculated as [lyso-Gb3]*age): MSSI (r2 = 0.88, p< 0.0001), LVMI (r2 = 0.59, p< 0.005), and GFR (r2 = 0.75, p = 0.0001). CONCLUSION: These results demonstrate that the N215S mutation results in a late onset phenotype involving the heart and other organs. Correlations between clinical manifestations and plasma lyso-Gb3 variations in this group suggest a Fabry-relevant disease mechanism for the heterogeneity observed in this group.


Subject(s)
Fabry Disease/pathology , alpha-Galactosidase/genetics , Adolescent , Adult , Age of Onset , Aged , Aged, 80 and over , Carboxymethylcellulose Sodium , Fabry Disease/genetics , Female , Genetic Association Studies , Humans , Male , Middle Aged , Mutation, Missense/genetics , Retrospective Studies , Young Adult
2.
Plant Cell Physiol ; 56(9): 1786-97, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26187898

ABSTRACT

The roles of non-cellulosic polysaccharides in cotton fiber development are poorly understood. Combining glycan microarrays and in situ analyses with monoclonal antibodies, polysaccharide linkage analyses and transcript profiling, the occurrence of heteromannan and heteroxylan polysaccharides and related genes in developing and mature cotton (Gossypium spp.) fibers has been determined. Comparative analyses on cotton fibers at selected days post-anthesis indicate different temporal and spatial regulation of heteromannan and heteroxylan during fiber development. The LM21 heteromannan epitope was more abundant during the fiber elongation phase and localized mainly in the primary cell wall. In contrast, the AX1 heteroxylan epitope occurred at the transition phase and during secondary cell wall deposition, and localized in both the primary and the secondary cell walls of the cotton fiber. These developmental dynamics were supported by transcript profiling of biosynthetic genes. Whereas our data suggest a role for heteromannan in fiber elongation, heteroxylan is likely to be involved in the regulation of cellulose deposition of secondary cell walls. In addition, the relative abundance of these epitopes during fiber development varied between cotton lines with contrasting fiber characteristics from four species (G. hirsutum, G. barbadense, G. arboreum and G. herbaceum), suggesting that these non-cellulosic polysaccharides may be involved in determining final fiber quality and suitability for industrial processing.


Subject(s)
Cell Wall/metabolism , Cotton Fiber , Epitopes/metabolism , Mannans/metabolism , Xylans/metabolism , Biomechanical Phenomena , Cluster Analysis , Gene Expression Regulation, Plant , Genes, Plant , Microarray Analysis , Monosaccharides/analysis , Species Specificity
3.
Age Ageing ; 43(2): 289-92, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24305621

ABSTRACT

BACKGROUND: circulating measures of inflammatory markers, such as C-reactive protein (CRP) have been associated with an increased risk of future cognitive decline. However, the nature of the relationship among the very old (>75 years) is unclear. Cross-sectional evidence suggests that elevated CRP may even be protective in this age group. This study examines these associations longitudinally. METHODS: logistic regression was used to investigate the association between CRP and drop in cognitive performance (≥3 point change on the Mini-Mental State Examination) over a 4-year period in a population of 266 people, mean age 77 years. RESULTS: increased levels of CRP were associated with a decreased risk of a drop in cognitive performance; however, this association was only seen in those without an APOE e4 allele [odds ratio of decline per unit increase in ln(CRP) 0.57, P = 0.04]. The magnitude of the finding remained consistent after adjustment for cardiovascular confounders (smoking, drinking, MI, stroke, diabetes, education, medication and blood pressure). For those with an e4 allele, the relationship with longitudinal cognitive decline was neither statistically significant nor in a consistent direction after controlling for acute inflammation. CONCLUSIONS: this study strengthens previous cross-sectional findings and shows elevated levels of CRP to be linked to a decreased risk of longitudinal cognitive decline in the very old. However, as with prior analyses, this was only observed in those not carrying an APOE e4 allele. Future work on larger APOE e4 allele carrying samples is required to determine the nature of the association in this population.


Subject(s)
Aging , Apolipoprotein E4/genetics , C-Reactive Protein/metabolism , Cognition Disorders/etiology , Cognition , Inflammation Mediators/blood , Age Factors , Aged , Aged, 80 and over , Aging/blood , Aging/genetics , Aging/immunology , Aging/psychology , Biomarkers/blood , Cognition Disorders/blood , Cognition Disorders/genetics , Cognition Disorders/immunology , Cognition Disorders/psychology , Female , Genotype , Humans , Logistic Models , Longitudinal Studies , Male , Odds Ratio , Phenotype , Prospective Studies , Psychiatric Status Rating Scales , Risk Factors , Time Factors , Up-Regulation
4.
Int J Obes (Lond) ; 37(12): 1571-8, 2013 Dec.
Article in English | MEDLINE | ID: mdl-23567927

ABSTRACT

OBJECTIVE: The Medifast 5 & 1 Plan (MD) is a portion-controlled, nutritionally-balanced, low-fat weight-loss plan. We studied the effects of MD compared with a reduced-energy, food-based diet (FB) on body weight, waist circumference, fat mass and other measures in adults. DESIGN: We conducted a two-parallel-arm, randomized, controlled trial comparing MD to FB over 52 weeks. A total of 120 men and women aged 19-65 years with BMI 35 and 50 kg m(-2) were randomized to MD (n=60) or FB (n=60). Follow-up included a 26-week weight-loss phase and 26-week weight-maintenance phase. Anthropometric, body composition, biochemical and appetite/satiety measures were performed at baseline and at 26 and 52 weeks. An intention-to-treat, linear mixed models analysis was the primary analysis. RESULTS: Fifty MD subjects (83.3%) and 45 FB subjects (75.0%) completed the study on assigned treatment. At 26 weeks, race-adjusted mean weight loss was 7.5 kg in MD subjects vs 3.8 kg in FB subjects (P=0.0002 for difference); reduction in waist circumference was 5.7 cm in MD vs 3.7 cm in FB (P=0.0064); and fat mass loss was 6.4 kg in MD vs 3.7 kg in FB (P=0.0011). At 52 weeks, the corresponding reductions were 4.7 vs 1.9 kg (P=0.0004); 5.0 vs 3.6 cm (P=0.0082); and 4.1 vs 1.9 kg (P=0.0019) in MD and FB subjects, respectively. CONCLUSION: In obese adults, MD resulted in significantly greater reductions in body weight and fat compared with an FB diet for 1 year after randomization.


Subject(s)
Diet, Reducing/methods , Energy Intake , Obesity/diet therapy , Satiation , Waist Circumference , Weight Loss , Weight Reduction Programs/methods , Adult , Aged , Body Weight , Diet, Fat-Restricted , Female , Humans , Male , Middle Aged , Obesity/epidemiology , Patient Compliance/statistics & numerical data , Patient Satisfaction/statistics & numerical data , Program Evaluation , Treatment Outcome
5.
Blood Cells Mol Dis ; 50(3): 212-7, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23219328

ABSTRACT

Gaucher disease (GD) is an inherited enzyme deficiency characterised by progressive cytopenias, hepatosplenomegaly and destructive bone disease. It is diagnosed by demonstration of beta glucosidase deficiency but may be suspected in presence of abnormal storage cells on tissue biopsy. Specific treatment is available in the form of enzyme replacement (ERT) and is effective in reversing many disease features. Delayed treatment has been associated with increased disease complications. This retrospective review of a single centre cohort of 86 patients was undertaken to ascertain if the diagnostic journey had improved since the introduction of ERT and commissioning of services. Fifty-six percent of patients presented primarily with features related to thrombocytopenia or splenomegaly with a median time from symptom onset to diagnosis of 2years (range 0.5-26years), 19% experiencing delays of 5 or more years. Seventy-five percent of patients were diagnosed by haematologists, 68% following an abnormal bone marrow biopsy. Raised serum ACE levels, low HDL cholesterol and raised ferritin were identified as prevalent laboratory abnormalities at the time of diagnosis. These features, coupled with the relative preservation of haemoglobin and white cell counts compared to the platelet count, help identify patients presenting to haematologists with a possible diagnosis of GD earlier in the diagnostic pathway.


Subject(s)
Gaucher Disease/diagnosis , Adolescent , Adult , Aged , Child , Child, Preschool , Diagnosis, Differential , Diagnostic Tests, Routine , Female , Gaucher Disease/pathology , Humans , Infant , Infant, Newborn , Male , Middle Aged , Phenotype , Retrospective Studies , Young Adult
6.
Carbohydr Polym ; 89(1): 213-21, 2012 Jun 05.
Article in English | MEDLINE | ID: mdl-24750626

ABSTRACT

Carbohydrate-binding modules (CBMs) are a set of tools that can be used as molecular probes for studying plant cell walls and cellulose-based substrates. CBMs from enzymes of bacterial and fungal origin present a range of recognition capabilities for crystalline and amorphous cellulose. Here cellulose-directed CBMs have been used to visualize and quantify crystallinity changes in cellulose II-based polymers following NaOH treatment. Cellulose II polymers used were in the form of lyocell fibers, which are derived from eucalyptus wood pulp. The supramolecular structure, morphology, and existence of 'skin-core' model in the fiber were examined using CBM-labeling techniques. Changes in cellulose crystallinity showed maxima at 3.33 mol dm(-3) NaOH (under treatment conditions of 49 Nm(-1) at 25 °C) and 4.48 mol dm(-3) NaOH (under treatment conditions of 147 Nm(-1) at 40 °C); CBM methods were also suitable for quantifying changes within amorphous regions. Quantification of crystallinity changes using CBM labeling techniques was achieved in combination with image analysis, which was shown to reflect the same crystallinity changes as measured using ATR-FTIR methods. It was demonstrated that CBM-labeling techniques were able to validate the proposed 'skin-core' model of lyocell fibers, comprising a semi-permeable fiber skin and a porous core.


Subject(s)
Cellulose/chemistry , Crystallization , Sodium Hydroxide/chemistry , Spectroscopy, Fourier Transform Infrared
7.
Biomacromolecules ; 12(11): 4121-6, 2011 Nov 14.
Article in English | MEDLINE | ID: mdl-21981266

ABSTRACT

Cotton fiber cellulose is highly crystalline and oriented; when native cellulose (cellulose I) is treated with certain alkali concentrations, intermolecular hydrogen bonds are broken and Na-cellulose I is formed. At higher alkali concentrations Na-cellulose II forms, wherein intermolecular and intramolecular hydrogen bonds are broken, ultimately resulting in cellulose II polymers. Crystallinity changes in cotton fibers were observed and assigned using attenuated total reflectance Fourier transform infrared (ATR FT-IR) spectroscopy and X-ray diffraction (XRD) subsequent to sodium hydroxide treatment and compared with an in situ protein-binding methodology using cellulose-directed carbohydrate-binding modules (CBMs). Crystallinity changes observed using CBM probes for crystalline cellulose (CBM2a, CBM3a) and amorphous cellulose (CBM4-1, CBM17) displayed close agreement with changes in crystallinity observed with ATR-FTIR techniques, but it is notable that crystallinity changes observed with CBMs are observed at lower NaOH concentrations (2.0 mol dm(-3)), indicating these probes may be more sensitive in detecting crystallinity changes than those calculated using FTIR indices. It was observed that the concentration of NaOH at which crystallinity changes occur as analyzed using the CBM labeling techniques are also lower than those observed using X-ray diffraction techniques. Analysis of crystallinity changes in cellulose using CBMs offers a new and advantageous method of qualitative and quantitative assessment of changes to the structure of cellulose that occur with sodium hydroxide treatment.


Subject(s)
Cellulose/chemistry , Peptide Fragments/chemistry , Binding Sites , Crystallization , Gossypium/chemistry , Hydrogen Bonding , Protein Binding , Protein Structure, Tertiary , Sodium Hydroxide/chemistry , Spectroscopy, Fourier Transform Infrared , X-Ray Diffraction
8.
Lupus ; 20(14): 1547-50, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21993386

ABSTRACT

Pure red cell aplasia (PRCA) is a recognized but rare complication of systemic lupus erythematosus (SLE) and is characterized by the near absence of red blood cell precursors in the bone marrow but with normal megakaryocyte and granulocytes. We report a novel case of acquired PRCA occurring simultaneously with immune thrombocytopenia in the context of active SLE. Both syndromes were refractory to conventional treatment but responded to rituximab and cyclophosphamide.


Subject(s)
Lupus Erythematosus, Systemic/complications , Red-Cell Aplasia, Pure/etiology , Thrombocytopenia/immunology , Antibodies, Monoclonal, Murine-Derived/therapeutic use , Antirheumatic Agents/therapeutic use , Cyclophosphamide/therapeutic use , Female , Glucocorticoids/therapeutic use , Humans , Lupus Erythematosus, Systemic/drug therapy , Red-Cell Aplasia, Pure/drug therapy , Rituximab , Thrombocytopenia/drug therapy , Young Adult
9.
Plant J ; 64(2): 191-203, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20659281

ABSTRACT

How the diverse polysaccharides present in plant cell walls are assembled and interlinked into functional composites is not known in detail. Here, using two novel monoclonal antibodies and a carbohydrate-binding module directed against the mannan group of hemicellulose cell wall polysaccharides, we show that molecular recognition of mannan polysaccharides present in intact cell walls is severely restricted. In secondary cell walls, mannan esterification can prevent probe recognition of epitopes/ligands, and detection of mannans in primary cell walls can be effectively blocked by the presence of pectic homogalacturonan. Masking by pectic homogalacturonan is shown to be a widespread phenomenon in parenchyma systems, and masked mannan was found to be a feature of cell wall regions at pit fields. Direct fluorescence imaging using a mannan-specific carbohydrate-binding module and sequential enzyme treatments with an endo-ß-mannanase confirmed the presence of cryptic epitopes and that the masking of primary cell wall mannan by pectin is a potential mechanism for controlling cell wall micro-environments.


Subject(s)
Cell Wall/metabolism , Mannans/metabolism , Animals , Antibodies, Monoclonal/metabolism , Esterification , Fluorescent Antibody Technique, Direct , Magnoliopsida , Male , Pectins/metabolism , Pinus , Rats , Rats, Wistar , beta-Mannosidase/metabolism
10.
Phys Med Biol ; 53(19): 5371-83, 2008 Oct 07.
Article in English | MEDLINE | ID: mdl-18765891

ABSTRACT

Optical computed tomography (optical-CT) and optical-emission computed tomography (optical-ECT) are new techniques for imaging the 3D structure and function (including gene expression) of whole unsectioned tissue samples. This work presents a method of improving the quantitative accuracy of optical-ECT by correcting for the 'self'-attenuation of photons emitted within the sample. The correction is analogous to a method commonly applied in single-photon-emission computed tomography reconstruction. The performance of the correction method was investigated by application to a transparent cylindrical gelatin phantom, containing a known distribution of attenuation (a central ink-doped gelatine core) and a known distribution of fluorescing fibres. Attenuation corrected and uncorrected optical-ECT images were reconstructed on the phantom to enable an evaluation of the effectiveness of the correction. Significant attenuation artefacts were observed in the uncorrected images where the central fibre appeared approximately 24% less intense due to greater attenuation from the surrounding ink-doped gelatin. This artefact was almost completely removed in the attenuation-corrected image, where the central fibre was within approximately 4% of the others. The successful phantom test enabled application of attenuation correction to optical-ECT images of an unsectioned human breast xenograft tumour grown subcutaneously on the hind leg of a nude mouse. This tumour cell line had been genetically labelled (pre-implantation) with fluorescent reporter genes such that all viable tumour cells expressed constitutive red fluorescent protein and hypoxia-inducible factor 1 transcription-produced green fluorescent protein. In addition to the fluorescent reporter labelling of gene expression, the tumour microvasculature was labelled by a light-absorbing vasculature contrast agent delivered in vivo by tail-vein injection. Optical-CT transmission images yielded high-resolution 3D images of the absorbing contrast agent, and revealed highly inhomogeneous vasculature perfusion within the tumour. Optical-ECT emission images yielded high-resolution 3D images of the fluorescent protein distribution in the tumour. Attenuation-uncorrected optical-ECT images showed clear loss of signal in regions of high attenuation, including regions of high perfusion, where attenuation is increased by increased vascular ink stain. Application of attenuation correction showed significant changes in an apparent expression of fluorescent proteins, confirming the importance of the attenuation correction. In conclusion, this work presents the first development and application of an attenuation correction for optical-ECT imaging. The results suggest that successful attenuation correction for optical-ECT is feasible and is essential for quantitatively accurate optical-ECT imaging.


Subject(s)
Hypoxia-Inducible Factor 1/analysis , Image Enhancement/methods , Tomography, Optical/methods , Neoplasms/diagnosis , Phantoms, Imaging , Probability , Reproducibility of Results , Sensitivity and Specificity , Transplantation, Heterologous
12.
Neuron ; 49(1): 41-53, 2006 Jan 05.
Article in English | MEDLINE | ID: mdl-16387638

ABSTRACT

Although mutations in the human doublecortin gene (DCX) cause profound defects in cortical neuronal migration, a genetic deletion of Dcx in mice produces a milder defect. A second locus, doublecortin-like kinase (Dclk), encodes a protein with similar "doublecortin domains" and microtubule stabilization properties that may compensate for Dcx. Here, we generate a mouse with a Dclk mutation that causes no obvious migrational abnormalities but show that mice mutant for both Dcx and Dclk demonstrate perinatal lethality, disorganized neocortical layering, and profound hippocampal cytoarchitectural disorganization. Surprisingly, Dcx(-/y);Dclk(-/-) mutants have widespread axonal defects, affecting the corpus callosum, anterior commissure, subcortical fiber tracts, and internal capsule. Dcx/Dclk-deficient dissociated neurons show abnormal axon outgrowth and dendritic structure, with defects in axonal transport of synaptic vesicle proteins. Dcx and Dclk may directly or indirectly regulate microtubule-based vesicle transport, a process critical to both neuronal migration and axon outgrowth.


Subject(s)
Axons/physiology , Cell Movement/physiology , Microtubule-Associated Proteins/genetics , Neurons/physiology , Neuropeptides/genetics , Protein Serine-Threonine Kinases/genetics , Animals , Animals, Newborn , Axons/ultrastructure , Brain/abnormalities , Brain/embryology , Congenital Abnormalities/genetics , Congenital Abnormalities/mortality , Congenital Abnormalities/pathology , Dendrites/ultrastructure , Doublecortin Domain Proteins , Doublecortin Protein , Doublecortin-Like Kinases , Embryo, Mammalian/metabolism , Embryo, Mammalian/pathology , Mice , Mice, Knockout , Mice, Mutant Strains , Microtubule-Associated Proteins/physiology , Neocortex/embryology , Nerve Tissue Proteins/deficiency , Neuropeptides/physiology , Protein Serine-Threonine Kinases/physiology , RNA, Messenger/metabolism , Synaptic Vesicles/metabolism , Tissue Survival
13.
Cochrane Database Syst Rev ; (2): CD003862, 2005 Apr 18.
Article in English | MEDLINE | ID: mdl-15846684

ABSTRACT

BACKGROUND: Road crashes are a prime cause of death and disability and red-light running is a common cause of crashes at signalised intersections. Red-light cameras are increasingly used to promote compliance with traffic signals. Manual enforcement methods are resource intensive and high risk, whereas red-light cameras can operate 24 hours a day and do not involve high-speed pursuits. OBJECTIVES: To quantify the impact of red-light cameras on the incidence and severity of road crashes and casualties, and the incidence of red-light violations. SEARCH STRATEGY: We searched the following electronic databases: TRANSPORT (NTIS, TRIS, IRRD,TRANSDOC), Cochrane Injuries Group Specialised Register, Cochrane Controlled Trials Register, MEDLINE, EMBASE and the Australian Transport Index. We checked the reference lists of relevant papers and contacted research and advocacy organisations. SELECTION CRITERIA: Randomised or quasi-controlled trials and controlled before-after studies of red-light cameras. For crash impact evaluation, the before and after periods each had to be at least one year in length. For violation studies, the after period had to occur at least one year after camera installation. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data on study type, characteristics of camera and control areas, and data collection period. Before-after data were collected on number of crashes by severity, collision type, deaths and injuries, and red-light violations. Rate ratio was calculated for each study. Where there was more than one, rate ratios were pooled to give an overall estimate, using a generic inverse variance method and a random-effects model. MAIN RESULTS: No randomised controlled trials were identified but 10 controlled before-after studies from Australia, Singapore and the USA met our inclusion criteria. We grouped them according to the extent to which they adjusted for regression to the mean (RTM) and spillover effects. Total casualty crashes: the only study that adjusted for both reported a rate ratio of 0.71 (95% CI to 0.55, 0.93); for three that partially adjusted for RTM but failed to consider spillover, rate ratio was 0.87 (95% CI to 0.77, 0.98); one that made no adjustments had a rate ratio of 0.80 (95% CI 0.58 to 1.12). Right-angle casualty crashes: rate ratio for two studies that partially addressed RTM was 0.76 (95% CI 0.54 to 1.07). Total crashes: the study addressing both RTM and spillover reported a rate ratio of 0.93 (95% CI 0.83 to 1.05); one study that partially addressed RTM had a rate ratio of 0.92 (95% CI 0.73 to 1.15); the pooled rate ratio from the five studies with no adjustments was 0.74 (95% CI 0.53 to 1.03). Red-light violations: one study found a rate ratio of 0.53 (95% CI 0.17 to 1.66). AUTHORS' CONCLUSIONS: Red-light cameras are effective in reducing total casualty crashes. The evidence is less conclusive on total collisions, specific casualty collision types and violations, where reductions achieved could be explained by the play of chance. Most evaluations did not adjust for RTM or spillover, affecting their accuracy. Larger and better controlled studies are needed.


Subject(s)
Accident Prevention/methods , Accidents, Traffic/prevention & control , Photography/methods , Accident Prevention/instrumentation , Controlled Clinical Trials as Topic , Humans , Photography/instrumentation
14.
J Forensic Sci ; 37(6): 1652-5, 1992 Nov.
Article in English | MEDLINE | ID: mdl-1453173

ABSTRACT

An immunoblotting method for phenotyping haptoglobin in serum and bloodstains has been developed. Haptoglobin isoproteins were separated by polyacrylamide gradient gel electrophoresis and then transferred to nitrocellulose by electroblotting. The use of 1 mm gels facilitated more rapid and effective transfer than conventional 3 mm thick gels. Nitrocellulose blots were developed by double antibody enzyme immunoassay. The detection limit for serum and bloodstains was improved 16 times compared to conventional staining using O-tolidine. The method could detect haptoglobin phenotypes from 0.001 microliter of whole blood. This detection limit is approximately 8 times lower than that of group specific-component analysis by immunoblotting.


Subject(s)
Blood Stains , Haptoglobins/genetics , Immunoblotting/methods , Animals , Cats , Dogs , Goats , Horses , Humans , Phenotype , Sheep
15.
Forensic Sci Int ; 47(2): 173-80, 1990 Sep.
Article in English | MEDLINE | ID: mdl-2227732

ABSTRACT

The application of a polyacrylamide gel isoelectric focusing (PAGIEF) and immunoblotting procedure for the identification of native alpha 2HS-glycoprotein (AHSG) in routine casework blood stains has produced reportable results on 57.2% of samples. This reporting rate is lower than that for group specific component (GC) (83.8%) and phosphoglucomutase (PGM 1) (72.8%) phenotyping of the same samples. Blood stain samples were desialyzed with 1 U/ml neuraminidase, overnight at room temperature prior to PAGIEF in gels containing pharmalyte pH 5-6 and 2.5 M urea. Simple AHSG patterns were developed by immunoblotting. This procedure was five times as sensitive as the native AHSG method and desialyzation was reproducible over a range of incubation times and neuraminidase concentrations. The application of the desialyzed AHSG analysis to routine casework samples has resulted in a significant increase in the number of reportable results (762 reported out of 1027 samples). This reporting rate (74.2%) compares favourably with that for GC (79.1%) and PGH 1 (69.6%) phenotyping of the same samples. The three AHSG alleles (AHSG*1, 2 and 3) are clearly resolved after sample desialyzation and separation in gels containing pharmalyte pH 5-6 and 2.5 M urea. The sensitivity of desialyzed AHSG phenotyping approaches that of GC and this technique is worthy of inclusion in blood stain screening protocols of forensic laboratories in regions where the population has a limited range of rare AHSG alleles.


Subject(s)
Blood Proteins/analysis , Blood Stains , Humans , Immunoblotting/methods , Isoelectric Focusing/methods , Phenotype , alpha-2-HS-Glycoprotein
16.
J Forensic Sci Soc ; 29(5): 325-30, 1989.
Article in English | MEDLINE | ID: mdl-2584948

ABSTRACT

A sensitive immunoblotting procedure has been applied to the detection of alpha-2-HS-glycoprotein (A2HS) phenotypes from control and casework bloodstains. A2HS phenotypes were separated by thin layer polyacrylamide gel isoelectric focusing (PAGIEF) in gels containing Pharmalyte pH 4.2-4.9. After transfer to nitrocellulose by a rapid capillary blot, the A2HS phenotypes were developed using a double antibody enzyme-immunoassay. The evaluation of A2HS phenotyping of casework material was undertaken in parallel with phosphoglucomutase (PGM) phenotyping by PAGIEF. A total of 598 water extracts from casework bloodstains have been tested. Positive results were obtained in 84% and 75% of samples for PGM and A2HS respectively. The A2HS gene frequencies A2HS*1 = 0.6420, A2HS*2 = 0.3530, and A2HS*3 = 0.0050 were determined from a survey of 1000 people in Brisbane.


Subject(s)
Blood Proteins/analysis , Blood Stains , Alleles , Blood Proteins/genetics , Gene Frequency , Humans , Immunoblotting , Isoelectric Focusing , Phenotype , alpha-2-HS-Glycoprotein
17.
J Forensic Sci Soc ; 29(4): 243-8, 1989.
Article in English | MEDLINE | ID: mdl-2529346

ABSTRACT

The performance of the polymorphic marker systems group-specific component (GC), phosphoglucomutase-1 (PGM), alpha-2-HS-glycoprotein (A2HS), haptoglobin (Hp), and erythrocyte acid phosphatase (EAP) was evaluated on control bloodstains. The major factors considered were: sensitivity of the test system; stability of the marker; laboratory economics of each test; and distinguishing power (Dp) of the system. GC was considered to be the most suitable marker for routine screening because of its high stability and Dp, and the sensitivity of the immunoblotting detection method. PGM and A2HS were the next most valuable markers followed by Hp. EAP could only be considered useful when large amounts of relatively fresh bloodstain were available.


Subject(s)
Acid Phosphatase/blood , Blood Proteins/analysis , Blood Stains , Phosphoglucomutase/blood , Vitamin D-Binding Protein/blood , Erythrocytes/enzymology , Haptoglobins/analysis , Humans , Phenotype , alpha-2-HS-Glycoprotein
18.
J Forensic Sci Soc ; 29(3): 197-205, 1989.
Article in English | MEDLINE | ID: mdl-2794913

ABSTRACT

A sensitive immunoblotting method for the routine detection of group-specific component (GC) from fresh serum, and from control and casework bloodstains has been developed. GC phenotypes were separated in a thin layer polyacrylamide gel by isoelectric focusing, transferred to nitrocellulose by a rapid capillary blotting procedure, and detected using a double antibody enzyme immunoassay. This method is capable of phenotyping 8 ng of GC extracted from bloodstains, a four-fold increase in sensitivity when compared to immunofixation and silverstaining. A total of 2424 casework bloodstains have been analysed and GC phenotypes identified in 78% of samples. The method is suitable for use in routine laboratories and is more sensitive than other methods for GC phenotyping of casework bloodstains.


Subject(s)
Blood Stains , Vitamin D-Binding Protein/genetics , Humans , Immunoblotting , Phenotype , Vitamin D-Binding Protein/blood
19.
Eur J Clin Pharmacol ; 35(4): 401-7, 1988.
Article in English | MEDLINE | ID: mdl-3197749

ABSTRACT

We have conducted a field trial of a pill-box containing a concealed electronic device for monitoring compliance in 23 consecutive adult out patients taking a rifampicin/isoniazid combination once daily. In 22 cases, the times when the box was opened were successfully recorded for the entire period (mean (SD) 26 (5) days) between successive clinic visits. In the other patient the record terminated after one week, a broken box being returned. Both totality of compliance (as assessed by box openings) and consistency of compliance (the proportion of the total number of intervals between openings which were of 22 to 26 h in length) were significantly greater in those studied in the intensive than in the maintenance phase of therapy. Patients may have taken the reduction in medication at the end of the intensive phase as signalling cure. A computer program has been developed to display the recorded data. This allowed the physician responsible to assimilate at a glance the patient's tablet-taking habits. In routine practice knowledge of the presence of the device may improve compliance and a discussion of the graphical display may prove of value in counselling.


Subject(s)
Isoniazid/therapeutic use , Patient Compliance , Rifampin/therapeutic use , Tuberculosis, Pulmonary/drug therapy , Adult , Drug Therapy, Combination , Electronics, Medical , Female , Humans , Male
20.
J Dairy Res ; 52(3): 379-89, 1985 Aug.
Article in English | MEDLINE | ID: mdl-2993389

ABSTRACT

Six dairy cows were treated before milkings with either oxytocin (Pitocin, 20 i.u.) or ACTH (Synacthen, 150 i.u.), principally to determine their effect on the ratio of citrate: lactoferrin concentrations in the milk. With ACTH treatment, after 3 d milk yield and citrate concentration decreased significantly, lactoferrin and bovine serum albumin (BSA) concentrations increased significantly. Somatic cell counts (SCC) increased temporarily in the milk of three of the cows which previously had greater than 100 000 cells/ml. Lactoferrin yield remained fairly constant but citrate yield was significantly reduced. The citrate: lactoferrin molar ratio decreased from 1373 to 606. With oxytocin treatment, after 4 d milk yield first increased and then significantly decreased, citrate concentration decreased significantly while there were no significant changes in lactoferrin or BSA concentration or in the yield of any other milk constituents. The citrate: lactoferrin molar ratio decreased from 1621 to 1301. There were no significant changes in SCC either during treatment or 4 d after treatment but there was a significant rise at 16 d after treatment. It was concluded that in lactating cows both hormones affected citrate and lactoferrin concentrations in the direction that would improve the antibacterial properties of milk, but that this was accompanied by adverse effects on milk secretion. The extent of the change was not sufficient to be likely to produce inhibition of coliform bacteria.


Subject(s)
Adrenocorticotropic Hormone/pharmacology , Cattle/metabolism , Citrates/metabolism , Lactoferrin/metabolism , Lactoglobulins/metabolism , Milk/drug effects , Oxytocin/pharmacology , Animals , Cell Count/veterinary , Citric Acid , Female , Milk/cytology , Milk/metabolism , Serum Albumin, Bovine/metabolism
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