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J Hopkins

Publications and source records attributed to J Hopkins.

At least 19 recordsLinked to original sources
PubMed2005

Seasonality of Old World screwworm myiasis in the Mesopotamia valley in Iraq.

Following the first recorded introduction of the Old World screwworm fly (OWS), Chrysomya bezziana Villeneuve (Diptera: Calliphoridae), into the Mesopotamia valley in Iraq in September 1996, cases of livestock myiasis caused by OWS developed a distinctly seasonal pattern. The annual cycle of clinical OWS cases is explained here on the basis of environmental variables that affect the different life-cycle stages of C. bezziana. This analysis suggests that low temperatures restricted pupal development during the winter, whereas the dispersal of adult flies was constrained by hot/dry summer conditions. A restricted number of OWS foci persisted throughout the year. In these foci, pupal development was fastest during the autumn months. In autumn, rapid multiplication, lasting several OWS generations, allowed subsequent adult fly dispersal across the valley floor during the winter. Hence, the monthly incidence of clinical OWS cases in livestock peaked during December-January and was lowest during July-August. In addition to temperature and humidity, vegetation cover played a role in OWS distribution. Hence the majority of OWS cases were clustered in the medium density type of vegetation [normalized difference vegetation index (NDVI) values of 0.2-0.4] along the main watercourses in the marshy Mesopotamia valley. Although sheep were the host most commonly infested by C. bezziana, local sheep density was not found to be a major factor in disease spread. Satellite imagery and the application of Geographical Information System (GIS) tools were found to be valuable in understanding the distribution of OWS in relation to vegetation and watercourses. The presence of screwworm in Iraq, at the perimeter of the intercontinental OWS distribution, may give rise to major seasonal flare-ups.

Animals↗
PubMed2005

Comparison of flow cytometric methods for the measurement of ZAP-70 expression in a routine diagnostic laboratory.

Chronic lymphocytic leukaemia (CLL) follows a variable clinical course with patient survival ranging from only a few years despite treatment, to several decades in patients who may never require clinical intervention. Determination of the mutational status of a patient's immunoglobulin heavy chain variable region (Ig V(H)) gene has been used to provide prognostic information, but this assay is not available in most laboratories. The discovery of the expression of the protein tyrosine kinase zeta-associated protein (ZAP)-70 in V(H)-unmutated CLL cases led to its proposal as a surrogate marker for V(H) status. This study investigated the measurement of ZAP-70 expression in CLL using different flow cytometric protocols. Two different antibodies and two different staining methods were compared. The Caltag ZAP-70 antibody and Fix & Perm kit were the easiest to use and were the most sensitive and specific combination, with 91% concordance between ZAP-70 expression and V(H) status. Three patients (9%) were discordant (two V(H) mutated/ZAP-70 positive, and one V(H) unmutated/ZAP-70 negative). No correlation existed between CD38 and either ZAP-70 expression or V(H) status. Measurement of ZAP-70 expression using the Caltag antibody/kit combination provides a standardized flow cytometric method that could be introduced into a routine CLL immunophenotyping panel in a clinical diagnostic laboratory.

Aged↗
PubMed2004

Correlation of structural development and differential expression of invasion-related molecules in schizonts of Plasmodium falciparum.

During asexual development Plasmodium schizonts undergo a series of complex biochemical and structural changes. Using tightly synchronized cultures of 2 P. falciparum lines (clone C10 and strain ITO4) for light microscopy and fluorescence imaging we monitored the timing and sequence of expression of proteins associated with invasion-related organelles. Antibodies to rhoptry, micronemal and dense granule proteins (Rhoptry Associated Protein 1, Apical Membrane Antigen 1, Erythrocyte Binding Antigen 175, Ring-infected Erythrocyte Surface Antigen) and to pellicle-associated proteins (Merozoite Surface Protein 1, PfMyosin-A) were used. Clone C10 developed faster than ITO4; this difference was also found in the timing of protein expression seen by immunofluorescence. Light microscopic data were combined with transmission electron microscopic analysis using serial sectioning of ITO4 schizonts to determine nuclear number and organellar development. Thus a timetable of schizont structural maturation was established. Generally, the timing of organelle-specific antigen expression correlates well with the ultrastructural data. Rhoptries are formed mainly between second and fourth nuclear divisions, micronemes between the end of the fourth nuclear division and merozoite separation from the residual body, while dense granules are generated mainly after the micronemes. PfAMA-1 appears in micronemes before EBA-175, suggesting micronemal heterogeneity.

Animals↗
PubMed2003

Human platelet glycoprotein VI function is antagonized by monoclonal antibody-derived Fab fragments.

Platelet interactions with adhesive ligands exposed at sites of vascular injury initiate the normal hemostatic response but may also lead to arterial thrombosis. Platelet membrane glycoprotein (GP)VI is a key receptor for collagen. Impairment of GPVI function in mice results in a long-term antithrombotic protection and prevents neointimal hyperplasia following arterial injury. On the other hand, GPVI deficiency in humans or mice does not result in serious bleeding tendencies. Blocking GPVI function may thus represent a new and safe antithrombotic approach, but no specific, potent anti-GPVI directed at the human receptor is yet available. Our aim was to produce accessible antagonists of human GPVI to evaluate the consequences of GPVI blockade. Amongst several monoclonal antibodies to the extracellular domain of human GPVI, one, 9O12.2, was selected for its capacity to disrupt the interaction of GPVI with collagen in a purified system and to prevent the adhesion of cells expressing recombinant GPVI to collagen and collagen-related peptides (CRP). While 9O12.2 IgGs induced platelet activation by a mechanism involving GPVI and Fc gamma RIIA, 9O12.2 Fab fragments completely blocked collagen-induced platelet aggregation and secretion from 5 microg mL-1 and fully prevented CRP-induced activation from 1.5 microg mL-1. 9O12.2 Fabs also inhibited the procoagulant activity of collagen-stimulated platelets and platelet adhesion to collagen in static conditions. Furthermore, 9O12.2 Fabs impaired platelet adhesion, and prevented thrombi formation under arterial flow conditions. We thus describe here for the first time a functional monoclonal antibody to human GPVI and demonstrate its effect on collagen-induced platelet aggregation and procoagulant activity, and on thrombus growth.

Antibodies, Monoclonal↗
PubMed2002

Granulocyte macrophage colony stimulating factor is elevated in alveolar macrophages from sheep naturally infected with maedi-visna virus and stimulates maedi-visna virus replication in macrophages in vitro.

Infection by maedi-visna virus, a lentivirus of sheep, leads to chronic inflammatory reactions of various tissues. In this report we have analysed the role of specific cytokines in the disease process. A significant increase in expression of interleukin-6, interleukin-10, granulocyte macrophage-colony stimulating factor (GM-CSF) and transforming growth factor-beta1 mRNA was observed in alveolar macrophages isolated from the lungs of naturally infected animals when compared with lungs of seronegative controls. Levels of GM-CSF mRNA expression in alveolar macrophages correlated with the presence of lung lesions, but there was no correlation of interleukin-10, interleukin-6, tumour necrosis factor-alpha and transforming growth factor-beta1 mRNA levels in alveolar macrophages from animals with pulmonary lesions. In vitro investigation showed that GM-CSF in the range 0.1-10 ng/ml induced a significant increase in viral p25 production after 7 days in acutely infected blood monocyte-derived macrophages. The production of p25 peaked between 7 and 14 days exposure to 10 ng/ml of GM-CSF. Quantitative polymerase chain reaction showed that the level of viral DNA in monocyte-derived macrophages was dose-dependent following GM-CSF treatment in the range 0.1-100 ng/ml after 7 days. Viral mRNA expression was also enhanced. These findings indicate a role for GM-CSF in the pathogenesis of lymphoid interstitial pneumonia in infected animals.

Animals↗
PubMed2002

A time for change.

Explore the source record for details and available documents.

Critical Care↗
PubMed2002

Cryotherapy and Transcutaneous Electric Neuromuscular Stimulation Decrease Arthrogenic Muscle Inhibition of the Vastus Medialis After Knee Joint Effusion.

OBJECTIVE: Arthrogenic muscle inhibition (AMI) is a presynaptic, ongoing reflex inhibition of joint musculature after distension or damage to the joint. The extent to which therapeutic interventions affect AMI is unknown. The purpose of this study was to verify that the vastus medialis (VM) is inhibited using the knee joint effusion model and to investigate the effects of cryotherapy and transcutaneous electric nerve stimulation (TENS) on AMI using this model. DESIGN AND SETTING: A 3 x 6 analysis of variance was used to compare Hoffmann-reflex data for treatment groups (cryotherapy, TENS, and control) across time (preinjection, postinjection, and 15, 30, 45, and 60 minutes after injection). SUBJECTS: Thirty neurologically sound volunteers (age = 21.8 +/- 2.4 years; height = 175.6 +/- 9.6 cm; mass = 71.5 +/- 13.3 kg) participated in this study. MEASUREMENTS: Hoffmann-reflex measurements were collected using a percutaneous stimulus to the femoral nerve and surface electromyography of the VM. RESULTS: Hoffmann-reflex measurements from the cryotherapy and TENS groups were greater than measurements from the control group at 15 and 30 minutes after injection. Measurements from the cryotherapy group were greater than for the TENS group, and measurements for the TENS group were greater than those for the control group at 45 minutes. At 60 minutes, the cryotherapy group measurements were greater than the TENS and control group measures. Measurements at 15, 30, 45, and 60 minutes after injection were reduced compared with the preinjection and postinjection measurements in the control group. Measurements in the cryotherapy group at 30, 45, and 60 minutes were greater than the preinjection, postinjection, and 15-minute data. No differences between time intervals existed in the TENS group. CONCLUSIONS: Artificial knee joint effusion results in VM inhibition. Cryotherapy and TENS both disinhibit the quadriceps after knee joint effusion, and cryotherapy further facilitates the quadriceps motoneuron pool. Cryotherapy treatment resulted in facilitation of the VM motoneuron pool during the post-treatment phase. The TENS treatment failed to disinhibit the VM motoneuron pool by 30 minutes postinjection.

Journal Article↗
PubMed2001

Crystal structure of PBP2x from a highly penicillin-resistant Streptococcus pneumoniae clinical isolate: a mosaic framework containing 83 mutations.

Penicillin-binding proteins (PBPs) are the main targets for beta-lactam antibiotics, such as penicillins and cephalosporins, in a wide range of bacterial species. In some Gram-positive strains, the surge of resistance to treatment with beta-lactams is primarily the result of the proliferation of mosaic PBP-encoding genes, which encode novel proteins by recombination. PBP2x is a primary resistance determinant in Streptococcus pneumoniae, and its modification is an essential step in the development of high level beta-lactam resistance. To understand such a resistance mechanism at an atomic level, we have solved the x-ray crystal structure of PBP2x from a highly penicillin-resistant clinical isolate of S. pneumoniae, Sp328, which harbors 83 mutations in the soluble region. In the proximity of the Sp328 PBP2x* active site, the Thr(338) --> Ala mutation weakens the local hydrogen bonding network, thus abrogating the stabilization of a crucial buried water molecule. In addition, the Ser(389) --> Leu and Asn(514) --> His mutations produce a destabilizing effect that generates an "open" active site. It has been suggested that peptidoglycan substrates for beta-lactam-resistant PBPs contain a large amount of abnormal, branched peptides, whereas sensitive strains tend to catalyze cross-linking of linear forms. Thus, in vivo, an "open" active site could facilitate the recognition of distinct, branched physiological substrates.

Alanine↗
PubMed2001

Linkage disequilibrium for schizophrenia at the chromosome 15q13-14 locus of the alpha7-nicotinic acetylcholine receptor subunit gene (CHRNA7).

The transmission/disequilibrium test was used for fine mapping of the linkage of schizophrenia to the chromosome 15q13-14 region, the site of a candidate gene, the alpha7 nicotinic acetylcholine receptor subunit gene (CHRNA7), in parent-child triads from the NIMH Schizophrenia Genetics Initiative families. This candidate gene was identified from neurobiological studies of deficits in schizophrenics of the inhibitory gating of the P50 auditory evoked potential. The neurobiological deficit was also used as a phenotype for subsequent linkage analysis. In the present study, significant genotype-wise disequilibrium (P < 0.007) was found at D15S165, a polymorphic simple sequence marker physically located within 1 megabase of both CHRNA7 and a partially duplicated, expressed sequence that includes exons 5-10 of CHRNA7. Replication of this result was found in an additional set of families. The results support this region as a chromosomal location involved in the genetic transmission of schizophrenia.

Chromosomes, Human, Pair 15↗
PubMed2001

Attachment, parenting, and marital dissatisfaction as predictors of disruptive behavior in preschoolers.

The aim of this study was to examine if an insecure coercive attachment pattern is associated with disruptive behavior in preschoolers, as well as to examine the concurrent and joint effects of attachment pattern, marital dissatisfaction, and ineffective parenting practices on disruptive behavior. Participants included 60 preschoolers and their mothers, recruited from three sites to ensure an adequate range of disruptive behavior. The Preschool Assessment of Attachment (Crittenden, 1992) was used to measure attachment pattern. Results of an analysis of variance revealed that children in the coercively attached dyads scored significantly higher on the measure of disruptive behavior than either the defended or secure children. Results of a hierarchical regression analysis indicated that the combination of a coercive pattern of attachment, marital dissatisfaction, and permissive parenting practices accounted for a significant proportion of the variance in disruptive behavior in preschoolers. These data suggest that a specific type of insecure attachment, a coercive pattern, is associated with disruptive behavior in preschoolers. Also, the data are consistent with previous findings of associations among marital dissatisfaction, ineffective parenting practices, and disruptive behavior.

Child Behavior Disorders↗
PubMed2001

Type 2 iodothyronin deiodinase transgene expression in the mouse heart causes cardiac-specific thyrotoxicosis.

Type 2 iodothyronine deiodinase (D(2)) catalyzes intracellular 3, 5, 3' triiodothyronine (T(3)) production from thyroxine (T(4)), and its messenger RNA mRNA is highly expressed in human, but not rodent, myocardium. The goal of this study was to identify the effects of D(2) expression in the mouse myocardium on cardiac function and gene expression. We prepared transgenic (TG) mice in which human D(2) expression was driven by the alpha-MHC promoter. Despite high myocardial D(2) activity, myocardial T(3) was, at most, minimally increased in TG myocardium. Although, plasma T(3) and T(4), growth rate as well as the heart weight was not affected by TG expression, there was a significant increase in heart rate of the isolated perfused hearts, from 284 +/-12 to 350 +/- 7 beats/min. This was accompanied by an increase in pacemaker channel (HCN2) but not alpha-MHC or SERCA II messenger RNA levels. Biochemical studies and (31)P-NMR spectroscopy showed significantly lower levels of phosphocreatine and creatine in TG hearts. These results suggest that even mild chronic myocardial thyrotoxicosis, such as may occur in human hyperthyroidism, can cause tachycardia and associated changes in high energy phosphate compounds independent of an increase in SERCA II and alpha-MHC.

Adenine Nucleotides↗
PubMed2001

Rapid deposition of wheat cell wall structural proteins in response to Fusarium-derived elicitors.

Two novel cell wall structural proteins of spring wheat (Triticum aestivum L. em Thell.). undergo rapid deposition in the cell wall matrix in a H(2)O(2)-dependent reaction after the elicitation of cultures with Fusarium graminearum (L.)-derived elicitor. The amino acid compositions of these proteins were remarkably similar and indicated that they were highly acidic (pI 3.8). These proteins contained 13--17% each of Gly, Glx and Ser with lesser amounts (6--8%) of Ala, Asx and Thr, and it has been suggested that they are known as glycine- and serine-rich proteins (GSRPs). SELDI-MS ionization spectra demonstrated that these proteins have low molecular masses of 8590 and 4292 Da. These results are discussed in relation to the possible role of these novel proteins in rapid, cell wall defensive reactions to pathogenic attack.

Amino Acids↗
PubMed2000

Effects of the depletion of CD8(+) T cells and monocytes on the proliferative responses of peripheral blood leucocytes from Trypanosoma evansi-infected sheep.

Sheep peripheral blood mononuclear cells and those depleted of CD8(+) T cells and/or monocytes were stimulated with polyclonal mitogens and specific antigens, and analysed by means of cell proliferation assay procedure to examine whether these cell populations are involved in Trypanosoma evansi-induced immunosuppression. The removal of CD8(+) T cells failed to normalize the proliferative responses of peripheral blood mononuclear cells from infected sheep to concanavalin A stimulation while the depletion of monocytes resulted in full and enhanced response, showing that macrophages are mainly responsible for the suppression. Although the depletion of CD8(+) T cells, monocytes or both restored the responses of the cells to lipopolysaccharide stimulation, the responsiveness of the undepleted cells to this mitogen was significantly higher from day 24 post infection (p<0.01). The results were discussed in relation to current known mechanisms of depressed lymphocyte proliferation in tsetse-transmitted African trypanosome infections.

Animals↗
PubMed2000

Sheep CD1 genes and proteins.

Interest in CD1 genes and proteins was initially stimulated by their close evolutionary and structural relationship to MHC class I molecules. The demonstration that CD1b and c molecules present novel non-peptide antigens to T-cells and play a role in protection against mycobacterial infection then focused attention on the functional role of CD1 proteins. Sheep possess at least seven CD1 genes, including CD1B, D and E, which is the most complex genetic arrangement identified so far in any animal. OvCD1B consists of at least three distinct genes, with the probability of limited polymorphism and the existence of splice variants. Most anti-sheep CD1-specific monoclonal antibodies react with OvCDlb and phenotypic and immunochemical data suggests the existence of two variants. CD1D genes have been identified in all species studied, suggesting a conserved role for CDld proteins across mammalian species. Presumptive evidence for the existence of OvCDIE has been obtained by NH2-terminal sequencing of protein precipitated by the mAb 20.27 (SBU-T6). Confirmatory evidence from gene cloning experiments is currently being sought. Collectively, these factors make the sheep CD1 family a highly relevant model for investigating the in vivo role of CD1 molecules. In this survey, the properties of monoclonal antibodies specific for sheep CD1, the cellular distribution and physicochemical characteristics of sheep CD1 molecules and the current state of knowledge on sheep CD1 genetics are reviewed.

Amino Acid Sequence↗
PubMed2000

Role of extensin peroxidase in tomato (Lycopersicon esculentum Mill.) seedling growth.

It is proposed that inhibition of extensin peroxidase activity leads to a less rigid cell wall and thus promotes cell expansion and plant growth. A low-molecular-weight inhibitor derived from the cell walls of suspension-cultured tomato cells was found to completely inhibit extensin peroxidase-mediated extensin cross-linking in vitro at a concentration of 260 microg/ml. The inhibitor had no effect upon guaiacol oxidation catalyzed by extensin peroxidase or horseradish peroxidase. We have demonstrated that the light-irradiated inhibition of plant growth may be partially offset by inhibition of endogenous extensin peroxidase activity. Overall plant growth was enhanced by up to 15% in the presence of inhibitor relative to control plants. Inhibitor-treated and illuminated tomato hypocotyls grew up to 15% taller than untreated controls. The inhibitor had no effect upon etiolated plants over a 15-d period, suggesting that only low levels of peroxidase-mediated cross-linking can be found in the cell walls of etiolated plants. SDS-PAGE/Western blots of ionically bound protein from both etiolated and illuminated hypocotyls identified a doublet at 57/58.5 kDa which is immunoreactive with antibodies raised to tomato extensin peroxidase. Levels of the 58.5-kDa protein, determined by SDS-PAGE, were at least threefold higher in illuminated tomato hypocotyls than in etiolated hypocotyls. Three fold higher levels of extensin peroxidase, elevated in-vitro extensin cross-linking activity and 15% higher levels of cross-linked, non-extractable extensin were observed in illuminated tomato hypocotyls compared with etiolated tomato hypocotyls. This suggests that white-light inhibition of tomato hypocotyl growth appears to be mediated, at least partially, by deposition of cell wall extensin, a process regulated by Mr-58,500 extensin peroxidase. Our results indicate that the contribution of peroxidase-mediated extensin deposition to plant cell wall architecture may have an important role in plant growth.

Cell Wall↗
PubMed2000

Quantitative analysis of maedi-visna virus DNA load in peripheral blood monocytes and alveolar macrophages.

Viral load may be an important indicator of disease progression in sheep infected with maedi-visna virus (MVV). To assess this variable accurately in MVV-infected sheep, a quantitative competitive-polymerase chain reaction (QC-PCR) was developed. A conserved region of the MVV pol gene was selected. The RT-PCR MVV pol product was cloned and mutagenised in vitro by PCR to produce a competitor template reduced in length from 217 to 192 bp, but which retained the original flanking MVV pol PCR primers. The competitor template was quantified accurately and in an optimised QC-PCR protocol serial dilutions of this template were co-amplified with known amounts of sample DNA. MVV DNA levels in peripheral blood monocytes and alveolar macrophages from MVV-infected sheep (n=12) were assessed by QC-PCR. Viral DNA load in alveolar macrophages was significantly higher than that in peripheral blood monocytes when the animals were compared overall. A comparison was also made between alveolar macrophages from the lungs of seropositive animals with or without histopathological evidence of pulmonary lesions. The load of MVV DNA in alveolar macrophages was low in sheep without histopathological evidence of lesions in the lung. In contrast, in alveolar macrophages from sheep with histopathological lesions in the lung, there was a significantly higher level of MVV DNA. The correlation of MVV load with pulmonary lesions suggests that infected alveolar macrophages play a key role in the pathogenesis of this lymphoid interstitial pneumonia.

Animals↗