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Biomedical subjects

M Galvin

Publications and source records attributed to M Galvin.

At least 19 recordsLinked to original sources

Evaluation of a spray-dried lacticin 3147 powder for the control of Listeria monocytogenes and Bacillus cereus in a range of food systems.

AIMS: The potential of a powdered preparation of the bacteriocin, lacticin 3147, was investigated for the inhibition of Listeria monocytogenes and Bacillus cereus. METHODS AND RESULTS: A 10% solution of reconstituted demineralized whey powder was fermented with Lactococcus lactis DPC3147 for the generation of a lacticin 3147 containing powdered product. A 99.9% reduction in L. monocytogenes numbers occurred in the presence of the lacticin 3147 powder within 2 h in natural yogurt, and an 85% reduction was observed in cottage cheese within the same time frame. Counts of B. cereus were reduced by 80% in soup, in the presence of 1% (w/w) lacticin 3147 powder, within 3 h. CONCLUSIONS: A powdered preparation of lacticin 3147 was effective for the control of Listeria and Bacillus in natural yogurt, cottage cheese and soup. SIGNIFICANCE AND IMPACT OF THE STUDY: The bioactive lacticin 3147 powder may find broad applications for control of Gram-positive pathogens/spoilage bacteria in a range of foods.

Bacillus cereus↗

Melanocortin 3 receptor (MC3R) gene variants in extremely obese women.

OBJECTIVE: Following several reports of linkage of obesity related phenotypes to human chromosome 20q we sought to determine whether variations of the melanocortin 3 receptor (MC3R) gene are associated with obesity. DESIGN: We screened the MC3R gene coding region and approximately 2 kb of 5' and 3' flanking sequences for DNA variants in unrelated extremely obese women and average weight controls using polymerase chain reaction (PCR) single strand conformation polymorphism (SSCP) analysis and DNA sequencing. SUBJECTS: 124 unrelated extremely obese women (body mass index, (BMI)>/=40 kg/m2) and 85 average weight controls (BMI<27 kg/m2). MEASUREMENTS: Radiation hybrid (RH) mapping was performed to localize the MC3R gene. 5' and 3' flanking sequences of MC3R gene were cloned. PCR-SSCP and DNA sequencing were used to detect mutations in the MC3R gene coding region and flanking sequences. RESULTS: RH mapping localized the MC3R gene to 20q13, between markers D20S100 and D20S149. 1083 bp 5' and 653 bp 3' flanking region of the MC3R gene were cloned. A missense mutation (+241, codon 81 ATT/GTT, Ile-->Val) was found in the MC3R coding region. Four more variants were detected in the 5' flanking sequence: -201(C-->G), -239 (A-->G), -762(A-->T) and -769(T-->C). Compared with controls, no significant allele frequency differences were found. Racial differences were found for the +241, -201, -239 and -762 polymorphisms. CONCLUSIONS: Several sequence variants were found in the MC3R gene coding region and in 5' flanking sequences. However, none of the variants were associated with obesity phenotypes. The linkage of extreme human obesity on 20q13 is likely caused by genes other than MC3R. International Journal of Obesity (2000) 24, 206-210

Adult↗

High prevalence of the very rare Wilson disease gene mutation Leu708Pro in the Island of Gran Canaria (Canary Islands, Spain): a genetic and clinical study.

The molecular basis of Wilson disease (WD), an autosomal recessive disorder, is the presence of mutations in the ATP7B gene, a copper transporting ATPase. Hospital records indicated a higher prevalence of WD (1 in 2,600) in some counties in the northeastern region of the island of Gran Canaria (Canary Islands, Spain) that was around 10-fold higher than that described for European populations (1 in 30,000). The ATP7B gene was analyzed for mutations in 24 affected subjects, revealing a high prevalence of the rare Leu708Pro mutation present in 12 homozygous and 7 heterozygous individuals. In these patients, who constitute one of the largest described cohorts of WD homozygotes, we found a variable clinical presentation of the disease, although the biochemical picture was homogenous and characteristic, thereby confirming that the Leu708Pro change is indeed a mutation associated with WD. Haplotype analysis of subjects homozygous for the Leu708Pro mutation showed a conserved shared region smaller than 1 centimorgan (cM), and the region of linkage disequilibrium between the Leu708Pro mutation and neighboring microsatellite markers extended approximately 4.6 cM. When comparing the amount of linkage disequilibrium versus genetic distance from the disease mutation, it was estimated that a common ancestral Leu708Pro chromosome may have been introduced in Gran Canaria over 56 generations ago, dating it back to pre-Hispanic times. The prevalence, and the tight geographical distribution of the Leu708Pro chromosome suggests that the Canary Islands can be considered a genetic isolate for linkage disequilibrium studies.

Adenosine Triphosphatases↗

Lacticin 3147 displays activity in buffer against gram-positive bacterial pathogens which appear insensitive in standard plate assays.

Lacticin 3147 is a broad-spectrum bacteriocin produced by Lactococcus lactis subsp. lactis DPC3147, which has been shown to be active against a range of food-borne bacteria. The reported inhibitory range for lacticin is extended to include methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecalis, penicillin-resistant Pneumococcus, Propionibacterium acne and Streptococcus mutans. This extended host range is not obvious from traditional agar plate-based methods, but reductions in bacterial cell numbers by up to 6 log10 cfu ml-1 was observed after 2 h in time-kill curve studies conducted in broth, suggesting that the bacteriocin may have potential as a therapeutic agent in the treatment of human infections.

Bacterial Proteins↗

Physical mapping of the CXC chemokine locus on human chromosome 4.

A physical map of the CXC chemokine locus on chromosome 4 has been constructed by PCR analysis and PFGE mapping of YAC clones. The genes for IL8, GRO1, PPBP, PF4, SCYB5 (ENA-78) and SCYB6 (GCP-2) have been co-localized on a 335-kb genomic fragment. The GRO2 and GRO3 genes did not map within this region and based on analysis of a YAC contig overlapping IL8 we speculate that GRO2 and GRO3 map downstream of this region. We have also assigned the novel CXC chemokine gene, SCYB9B (alias H174/betaR1) to chromosome 4q21, upstream and within 12 kb of INP10. Like INP10 and MIG, INP10 and SCYB9B are arranged in a head to tail manner. The chromosomal arrangement of these genes appears to reflect the evolution of this multigene family and supports the theory that it arose by gene duplication.

Biological Evolution↗

Fast and reliable screening of mutations in human tumors: use of multiple fluorescence-based long linker arm nucleotides assay (mf-LLA).

Human tumor samples were screened for point mutations by adapting a mobility-shift assay to automated DNA sizing. This screen identifies the type of point mutation and relative amount of mutated DNA sequences present in a sample. Test samples having known hypoxanthine-guanine phosphoribosyl transferase (hprt)/exon-3 sequence mutations were characterized by: (i) PCR amplification, (ii) fluorescent dye-primer extension with 36-atom linker derived deoxycytosine or deoxyuridine triphosphate and the remaining three natural nucleotides and (iii) sizing of the resulting fluorescently labeled modified strands, using an automated DNA sequencer. Routinely, a range of sizes is observed among the sequence variants of a single DNA target sequence. This is because nucleotide analogs are incorporated into DNA strands in a sequence-dependent manner, resulting in composition-dependent electrophoretic mobility. Thus, point mutations are identified as shifts in mobility between the fluorescently labeled modified strands of the control and test samples. The twenty different hprt/exon-3 single-base substitution mutations tested were easily identified, even at fourfold dilution with control DNA.

Animals↗

Development of a lacticin 3147-enriched whey powder with inhibitory activity against foodborne pathogens.

The broad-spectrum bacteriocin lacticin 3147, produced by Lactococcus lactis DPC3147, is inhibitory to a wide range of gram-positive food spoilage and pathogenic organisms. A 10% solution of demineralized whey powder was fermented with DPC3147 at a constant pH of 6.5. The fermentate was spray dried, and the resulting powder exhibited inhibitory activity. The ability of the lacticin 3147-enriched powder to inhibit Listeria monocytogenes Scott A and Staphylococcus aureus 10 was assessed in buffer at both acidic (pH 5) and neutral (pH 7) pH. In addition, the ability of the powder to inhibit L. monocytogenes Scott A in an infant milk formulation was assessed. Resuspension of approximately 10(8) midexponential phase L. monocytogenes Scott A cells in a 10% solution of the lacticin 3147-enriched powder resulted in a 1,000-fold reduction in viable cells at pH 5 and pH 7 after 3 h at 30 degrees C. In the case of S. aureus 10, resuspension of 2.5 x 10(7) midexponential phase cells in a 15% solution of the lacticin 3147-enriched powder at pH 5 resulted in only a 10-fold reduction in viable cell counts, compared with a 1,000-fold reduction at pH 7, following incubation for 3 h at 30 degrees C. The use of the lacticin 3147 powder in an infant milk formulation resulted in greater than a 99% kill of L. monocytogenes within 3 h at 30 degrees C. These results suggest that this bioactive lacticin 3147 food ingredient may find applications in many different foods, including those with pH close to neutrality.

Animals↗

"A satellite primer on tuberculosis:" a collaboration in distance education.

"A Satellite Primer on Tuberculosis" was offered as a distance-based certificate course on tuberculosis (TB) fundamentals to a national audience of over 5,000 individuals. The course was a collaborative effort of a school of public health, a state health department, and the Centers for Disease Control and Prevention. Instruction was provided through print-based self-study modules that were complemented by live, interactive satellite conferences. Course completers, over 70 percent of whom were nurses and employees of public health departments, scored significantly higher on a course posttest than on a pretest, and the vast majority felt the course provided valuable training.

Curriculum↗

Moralization of attachment: a fourth domain of conscience functioning.

OBJECTIVES: To define discrete developmental levels of understanding regarding the ways in which normal children and adolescents link remembered and current attachment experiences to their moral belief system and to study the correlation between this progression and previously identified stages of conscience conceptualization. METHOD: Using the moralization of attachment section from the semistructured Stilwell Conscience Interview, 132 normal volunteers between the ages of 5 and 17 years were individually interviewed. Analysis of the interviews resulted in five levels of understanding. RESULTS: By analyses of variance and covariance, the five attachment levels showed significant correlation with the five conceptualization stages. Conceptualization stage showed a stronger correlation than age. CONCLUSIONS: In normal development, moralization of attachment is a domain of conscience functioning which follows a five-level hierarchical developmental progression; first, the child's sense of security and empathic responsiveness become paired with a sense of moral obligation; caretaker rules are then incorporated; an understanding of how empathy modifies strict rule-following develops; idols and ideals are chosen that reflect earlier learning in attachment relationships; finally, a visualization of the self as moral standard-bearer or teacher unfolds.

Adolescent↗

Moral valuation: a third domain of conscience functioning.

OBJECTIVE: To assess development of moral valuation in normal children and adolescents, that is, how moral rules for living are derived and justified, and to examine the relationship of this progression with previously identified stages of conceptualization of conscience. METHOD: Using three semistructured questions from the Stilwell Conscience Interview, 132 normal volunteers between the ages of 5 and 17 years were assessed. All moral valuation responses were examined within three aspects of social reference: authority-derived, self-derived, and peer-derived. Each aspect was scaled for complexity into six anchored levels. RESULTS: The levels of all three aspects correlated positively with conceptualization stages as well as with each other. When the covariate, age, was taken into consideration, peer-derived valuation was significantly correlated with both age and stage. CONCLUSIONS: Moral valuation is a domain of conscience functioning in which moral rules and their justifications are socially referenced in relationship to authority, self, and peers. Anchored levels of these three aspects of moral valuation provide developmental guidelines for mental status examinations in patients between 5 and 17 years of age as well as providing criteria for future comparative studies in various diagnostic categories of psychopathology.

Adolescent↗

Peripheral blood stem cell transplantation in myeloma using CD34 selected cells.

We have performed nine CD34 selection procedures on peripheral blood stem cells harvested from eight patients with myeloma using the Cellpro avidin-biotin immunoaffinity column (Ceprate). They all received CVAMP chemotherapy to maximum response prior to mobilisation. Six of the patients have been transplanted using these cells, one receiving successive autografts. Median absolute cell numbers processed and retrieved were: 31.1 x 10(9) pre-column, 2.07 x 10(8) in the final product and 30.4 x 10(9) in the column waste. Mean CD34 positivity in the product was 49% (range 18.4-98) with a median CD34+ yield of 31.4% (range 21-37.8). IgH PCR was performed and seven of the eight patients were amplifiable. Of these, two were positive in the pre-column product and both of these were successfully purged with a negative result in the final, post-column product. Patients were transplanted with a median of 2.0 x 10(6) CD34+ cells/kg (range 1.5-9.4) following conditioning with melphalan 200 mg/m2. The mean time to recovery of neutrophils to > 0.5 x 10(9)/l and platelets to > 20 x 10(9)/l was 16 and 17 days, respectively. At a mean follow-up of 9 months, four of the six patients transplanted are alive, three of them in complete remission and one in a clinically stable relapse. One has died of disease relapse and one of progressive neurological problems the aetiology of which was uncertain but there was no sign of progression of their myeloma. We conclude that PBSCT using CD34 selected cells is safe and practical in myeloma following remission induction with CVAMP chemotherapy.

Antigens, CD34↗

Serum dopamine beta hydroxylase and maltreatment in psychiatrically hospitalized boys.

Fifty boys, hospitalized on a school-age and an adolescent unit in an intermediate length psychiatric hospital, were studied while off psychoactive medication to determine how serum dopamine beta hydroxylase (DBH) activity varies with different childhood maltreatment experiences. Childhood maltreatment was categorized according to onset (before 36 months old, between 36-72 months old and over 72 months old). Childhood maltreatment groups were compared with a group of psychiatrically hospitalized boys who had neither been abused nor neglected. Boys who were younger than 72 months at age of onset of maltreatment had significantly lower DBH activity than those who had experienced maltreatment later in childhood and those who had not been subjected to abuse or neglect. This difference appeared attributable to the DBH activity of school age (but not adolescent) boys who had been abused/neglected before 72 months. Boys with a principal diagnosis of conduct disorder solitary aggressive type had lower DBH activity than boys without this diagnosis regardless of whether or not they had been maltreated. Low serum DBH may be a biological sequela of maltreatment early in life that correlates with the development of conduct disorder solitary aggressive type in boys.

Adolescent↗

Practicing population-based care in an HMO: evaluation after 18 months.

OBJECTIVE: To determine whether population-based care in a primary care practice results in improvement in compliance with patient care guidelines. DESIGN: Time series analysis. SETTING: One primary care practice in Group Health Cooperative of Puget Sound (GHC). PARTICIPANTS: Approximately 1500 enrollees cared for by the practice. INTERVENTIONS: An ongoing approach to aid clinical planning at the level of the primary care team--population-based care--that depends on clinical guidelines, a computing system to provide epidemiologic data on guideline performance in the practice and reminders, and a process whereby the practice team analyzed and designs interventions for specific clinical problems. MAIN OUTCOME MEASURES: We compared compliance with practice guidelines for preventive care and chronic illness management at baseline and after 18 months in the intervention population with other patients in the same clinic and with patients in GHC as a whole. RESULTS: Compliance with breast cancer screening and colorectal cancer screening guidelines in the intervention population increased from baseline 32% and 18% respectively. These increases were significantly greater than in the remainder of the clinic or in GHC as a whole. CONCLUSIONS: The availability of practice-based data, clinical guidelines and a local intervention design process resulted in significant improvements in compliance with patient care guidelines.

Algorithms↗

Moral-emotional responsiveness: a two-factor domain of conscience functioning.

OBJECTIVE: The purpose of this study was to assess the progression in development of moral-emotional responsiveness in children and adolescents and to examine the relationship of this progression with previously identified stages of conceptualization of conscience. METHOD: Using three semistructured questions from the Stilwell Conscience Interview, 132 normal volunteers between the ages of 5 and 17 years were assessed regarding comprehension of their emotional responses to moral stimuli. RESULTS: Rational analysis of the responses identified six items; each item was scaled for complexity into five stages. Factor analysis of the six items revealed two factors: moral-emotional responsiveness 1 contained items relating to external anxiety, internal anxiety, and mood; more-emotional responsiveness 2 contained items relating to the restoration of psychophysiological equilibrium through the processes of reparation and healing. Differences between conceptualization stages, with the moral-emotional responsiveness factors serving as dependent variables, were accounted for by stage differences in age and the positive correlations between the moral-emotional responsiveness factors and age. CONCLUSIONS: Moral-emotional responsiveness is a two-factor domain of the conscience. The findings provide additional developmental guidelines for assessing conscience development and functioning both in clinical practice and in research.

Adolescent↗

Low dopamine-beta-hydroxylase: a biological sequela of abuse and neglect?

Twenty-one psychiatrically hospitalized boys were studied while off psychoactive medication to determine if conduct disorder, solitary type, and abuse or neglect experiences correlated with low levels of dopamine-beta-hydroxylase (DBH) activity. Preliminary results supported earlier findings that undersocialized types of conduct disorder in boys were correlated with low DBH activity. Possible or definite neglect or abuse before 36 months of age was correlated with low DBH activity. Abuse or neglect was not correlated with low DBH activity when time of occurrence was not specified. Low serum DBH may be a biological sequela of seriously disrupted attachment.

Adolescent↗

Conceptualization of conscience in normal children and adolescents, ages 5 to 17.

In order to determine how normal children and adolescents conceptualize their conscience, the Stilwell Conscience Interview was given to 125 normal subjects between the ages of 5 and 17. Responses to questions 1, 2 and 11 (including the drawing of conscience) were empirically analyzed, resulting in a five-stage developmental model. The responses were then randomly presented to two blinded raters to assign them to one of the five stages. Highly acceptable interrater reliability was found, Kappa = 0.90. The relationship of age to stage demonstrated a highly significant positive correlation, indicating that the conceptualization of conscience in normal development follows an invariant, hierarchical pattern of organization. A commentary regarding each stage is presented. The value of this conceptualization model as a comparative standard of normality in the clinical assessment of youngsters suffering from psychopathology is discussed relative to assessment, psychotherapy, and future research.

Adolescent↗