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Biomedical subjects

K Manning

Publications and source records attributed to K Manning.

At least 19 recordsLinked to original sources
PubMed2001

Expression of six expansin genes in relation to extension activity in developing strawberry fruit.

Expansins are proteins which have been demonstrated to induce cell wall extension in vitro. The identification and characterization of six expansin cDNAs from strawberry fruit, termed FaExp3 to FaExp7, as well as the previously identified FaExp2 is reported here. Analysis of expansin mRNAs during fruit development and in leaves, roots and stolons revealed a unique pattern of expression for each cDNA. FaExp3 mRNA was present at much lower levels than the other expansin mRNAs and was expressed in small green fruit and in ripe fruit. FaExp4 mRNA was present throughout fruit development, but was more strongly expressed during ripening. FaExp5 was the only clone to show fruit specific expression which was up-regulated at the onset of ripening. FaExp6 and FaExp7 mRNAs were present at low levels in the fruit with highest expression in stolon tissue. During fruit development FaExp6 had the highest expression at the white, turning and orange stages whereas expression of FaExp7 was highest in white fruit. The expression profiles of FaExp2 and FaExp5 in developing fruit were similar except that FaExp2 was induced at an earlier stage. Analysis of expansin protein by Western blotting using an antibody raised against CsExp1 from cucumber hypocotyls identified two bands of 29 and 31 kDa from developing fruit. Protein extracts from developing fruit were assayed for extension activity. Considerable rates of extension were observed with extracts from ripening fruit, but no extension was observed with protein from unripe green fruit. These results demonstrate the presence of at least six expansin genes in strawberry fruit and that during ripening the fruit acquires the ability to cause extension in vitro, characteristic of expansin action.

Amino Acid Sequence↗
PubMed2000

Risk factors for acquisition of vancomycin-resistant enterococci among patients on a renal ward during a community hospital outbreak.

BACKGROUND: During an outbreak of vancomycin-resistant enterococcal (VRE) infection and colonization at a community hospital in Indianapolis, Indiana, we performed a case-control study of patients on the hospital's renal unit to determine risk factors for acquisition of VRE among this potentially high-risk patient population. METHODS: Twenty-four renal patients with VRE colonization/infection (ie, case-patients) were compared by univariate and multivariate analyses with 29 renal patients with nosocomially acquired vancomycin-susceptible enterococcal infection and colonization (ie, controls). RESULTS: Age and length of hospitalization were similar between the VRE case-patients and the vancomycin-susceptible enterococcal control-patients, but case-patients had higher Acute Physiology and Chronic Health Evaluation II scores and received significantly greater numbers of antimicrobials and significantly more days of antimicrobials during the 60 days preceding the first positive enterococcal culture. In an assessment of the appropriateness of vancomycin use, one third of vancomycin orders were found to be inappropriate in both patient groups. CONCLUSIONS: Our data show that among renal patients, those who are severely ill and receive multiple and prolonged courses of antimicrobials are at greatest risk for acquiring VRE infection or colonization. The Centers for Disease Control and Prevention recommends that hospitals develop a comprehensive plan to prevent and control infection and colonization of patients with VRE. This plan should include prompt identification of affected patients, initiation of isolation precautions to prevent patient-to-patient transmission of VRE, and prudent use of antimicrobials, including vancomycin.

APACHE↗
PubMed1999

In vivo suppressor mutations correct a murine model of hereditary tyrosinemia type I.

Hereditary tyrosinemia type I and alkaptonuria are disorders of tyrosine catabolism caused by deficiency of fumarylacetoacetate hydrolase (FAH) and homogentisic acid dioxygenase (HGD), respectively. Tyrosinemia is a severe childhood disease that affects the liver and kidneys, but alkaptonuria is a more benign adult disorder in comparison. Because HGD is upstream of FAH in the tyrosine pathway, mice doubly mutant in both enzymes were found to be protected from the liver and renal damage of tyrosinemia as hypothesized. Mice mutant at the tyrosinemic locus but heterozygous for alkaptonuria spontaneously developed clonal nodules of functionally normal hepatocytes that were able to rescue the livers of some mice with this genotype. This phenotypic rescue was a result of an inactivating mutation of the wild-type homogentisic acid dioxygenase gene, thus presenting an example of an in vivo suppressor mutation in a mammalian model.

Acetoacetates↗
PubMed1999

Identification of the mutation in the alkaptonuria mouse model. Mutations in brief no. 216. Online.

Alkaptonuria (aku), an inborn error of metabolism caused by the loss of homogentisate 1,2-dioxygenase (HGD), has been described in a mouse model created by ethylnitrosourea mutagenesis but the mutation in these mice has not previously been identified. We used RT-PCR to amplify the Hgd cDNA from Hgd(aku)/Hgd(aku) mice. Two products shorter than the wild-type product were amplified. Restriction mapping and DNA sequencing were then used to identify the Hgd(aku) mouse mutation, found to be a single base change in a splice donor consensus sequence, causing exon skipping and frame-shifted products. This base change allowed us to create a non-radioactive genotyping assay for this allele.

Alkaptonuria↗
PubMed1999

Control of vancomycin-resistant enterococci at a community hospital: efficacy of patient and staff cohorting.

OBJECTIVE: To evaluate the efficacy of patient and staff cohorting to control vancomycin-resistant enterococci (VRE) at an Indianapolis community hospital. DESIGN: To interrupt transmission of VRE, a VRE point-prevalence survey of hospital inpatients was conducted, and VRE-infected or -colonized patients were cohorted on a single ward with dedicated nursing staff and patient-care equipment. To assess the impact of the intervention, staff compliance with contact isolation procedures was observed, and the VRE point-prevalence survey was repeated 2 months after the cohort ward was established. RESULTS: Following the establishment of the cohort ward, VRE prevalence among all hospitalized inpatients decreased from 8.1% to 4.7% (25 positive cultures among 310 patients compared to 13 positive cultures among 276 patients, P=.14); VRE prevalence among patients whose VRE status was unknown before cultures were obtained decreased from 5.9% to 0.8% (18 positive cultures among 303 patients compared to 2 positive cultures among 262 patients, P=.002); and observed staff-patient interactions compliant with published isolation recommendations increased (5 [22%] of 23 interactions compared to 36 [88%] of 41 interactions, P<.0001). CONCLUSIONS: Our data suggest that, in hospitals with endemic VRE or continued VRE transmission despite implementation of contact isolation measures, establishing a VRE cohort ward may be a practical and effective method to improve compliance with infection control measures and thereby to control epidemic or endemic VRE transmission.

Adult↗
PubMed1999

Etiology of acute lower respiratory tract infection in Central Australian Aboriginal children.

BACKGROUND: Aboriginal children in central Australia have attack rates for acute lower respiratory tract infection (ALRI) that are similar to those in developing countries. Although mortality rates are much lower than in developing countries, morbidity is high and ALRI is still the leading cause of hospitalization. However, there are no data on the etiology of ALRI in this population. METHODS: We prospectively studied 322 cases of ALRI in 280 Aboriginal children admitted to the hospital. Blood, urine and nasopharyngeal aspirate samples were examined for evidence of bacterial, viral and chlamydial infection. RESULTS: The combination of blood culture, viral studies and chlamydial serology provided at least 1 etiologic agent in 170 of 322 (52.5%) cases. Assays for pneumolysin immune complex and pneumolysin antibody increased etiologic diagnosis to 219 (68.0%). Blood cultures were positive in 6% but pneumolysin immune complex and pneumolysin antibody studies were positive in one-third of cases. Evidence of viral infection was present in 155 (48%) of cases compared with 12% in controls (P < 001). There were only 7 possible cases and 2 definite cases of Chlamydia trachomatis and 3 cases of Chlamydia pneumoniae. Coinfection was common in these children. CONCLUSION: These findings have implications for both standard treatment protocols and vaccine strategies. The high rate of coinfection may make it difficult to develop simple clinical predictors of bacterial infection. In the setting of a developed country with efficient patient evacuation services, management algorithms that focus on disease severity and need for hospital referral will be most useful to health staff in remote communities. Pneumococcal conjugate vaccines will be required to reduce the high attack rate of pneumococcal disease.

Australia↗
PubMed1998

Ex vivo hepatic gene therapy of a mouse model of Hereditary Tyrosinemia Type I.

Previously, this lab has reported the use of hepatocyte transplantation and in vivo gene therapy for the correction of a mouse model of Hereditary Tyrosinemia Type I (HT1). Here, we demonstrate repopulation of fumarylacetoacetate hydrolase (FAH)-deficient livers with cultured hepatocytes. Correction of the disease phenotype was achieved by retrovirally transducing cultured FAH- hepatocytes ex vivo, followed by transplantation and selective repopulation. Treated mice were phenotypically normal and had corrected plasma amino acid levels and liver function tests. Our results demonstrate that efficient hepatic repopulation using ex vivo genetically manipulated hepatocytes is feasible.

Amino Acid Metabolism, Inborn Errors↗
PubMed1998

Isolation of a set of ripening-related genes from strawberry: their identification and possible relationship to fruit quality traits.

The ripening of strawberry (Fragaria ananassa Duch.), a non-climacteric fruit, is a complex developmental process that involves many changes in gene expression. To understand how these changes relate to the biochemistry and composition of the fruit the specific genes involved have been examined. A high-quality cDNA library prepared from ripe strawberry fruit was differentially screened for ripening-related clones using cDNA from ripe and white fruits. From 112 up-regulated clones obtained in the primary screen, 66 differentially expressed clones were isolated from the secondary screen. The partial sequences of these cDNAs were compared with database sequences and 26 families of non-redundant clones were identified. Northern analysis confirmed that all of these cDNAs were ripening-enhanced. The expression of many of their corresponding genes was negatively regulated in auxin-treated fruit. These sequences, several of which are novel to fruits, encode proteins involved in key metabolic events including anthocyanin biosynthesis, cell wall degradation, sucrose and lipid metabolism, protein synthesis and degradation, and respiration. These findings are discussed in relation to the role of these genes in determining fruit quality characteristics.

Base Sequence↗
PubMed1998

Thoracocardiography: noninvasive monitoring of left ventricular stroke volume.

PURPOSE: Thoracocardiography noninvasively monitors global stroke volume by inductive plethysmographic recording of ventricular volume curves as previously validated by thermodilution. Our purpose was to investigate the potential of thoracocardiography to individually assess stroke volume of the left ventricle. We hypothesized that curves predominantly reflecting left ventricular volume could be obtained by recording waveforms from thoracocardiographic transducers placed at various levels around the chest, and by identifying their origin as the left ventricle if mean expiratory exceeded mean inspiratory stroke volumes during spontaneous breathing. MATERIALS AND METHODS: Stroke volumes obtained by thoracocardiography in normal subjects were compared beat by beat with estimates derived from simultaneous measurements of left ventricular cavity stroke area by echocardiography with automatic boundary detection. Changes in respiratory variations of stroke volumes were analyzed during spontaneous breathing at fixed rate and tidal volume, during mechanical ventilation, and resistive loaded breathing. RESULTS: In 170 comparisons of beat-by-beat stroke volumes, 89% of thoracocardiographic fell within +/-20% of echocardiographic estimates. Changes in tidal volume, resistive loaded breathing, and mechanical ventilation induced respiratory variations of thoracocardiographic derived stroke volumes consistent with the known effect of respiratory changes in intrapleural pressure on left ventricular stroke volumes. CONCLUSIONS: The results suggest that thoracocardiography noninvasively tracks changes in left ventricular stroke volumes. Their absolute value may also be monitored if an initial calibration by an independent technique, such as echocardiography, is performed.

Adult↗
PubMed1997

Gene expression in the pulp of ripening bananas. Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of in vitro translation products and cDNA cloning of 25 different ripening-related mRNAs.

mRNA was extracted from the pulp and peel of preclimacteric (d 0) bananas (Musa AAA group, cv Grand Nain) and those exposed to ethylene gas for 24 h and stored in air alone for a further 1 (d 2) and 4 d (d 5). Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of in vitro translation products from the pulp and peel of these fruits revealed significant up-regulation of numerous transcripts during ripening. The majority of the changes were initiated by d 2, with the level of these messages increasing during the remainder of the ripening period. Pulp tissue from d 2 was used for the construction of a cDNA library. This library was differentially screened for ripening-related clones using cDNA from d-0 and d-2 pulp by a novel microtiter plate method. In the primary screen 250 up- and down-regulated clones were isolated. Of these, 59 differentially expressed clones were obtained from the secondary screen. All of these cDNAs were partially sequenced and grouped into families after database searches. Twenty-five nonredundant groups of pulp clones were identified. These encoded enzymes were involved in ethylene biosynthesis, respiration, starch metabolism, cell wall degradation, and several other key metabolic events. We describe the analysis of these clones and their possible involvement in ripening.

Down-Regulation↗
PubMed1997

Does an increase in the sensitivity of serum thyrotropin assays reduce diagnostic costs for thyroid disease in the community?

Many authorities now advocate that the first-line assessment of thyroid function should be measurement of thyrotropin (TSH). The latest serum TSH assays (third generation) are more sensitive than the second generation but the reagents are more costly. We have examined whether overall assay reagent costs would be higher or lower with a third-generation assay, in a laboratory that serves a population of almost 500,000. In a prospective study over six weeks, 505 samples with a second-generation serum TSH less than 0.5 mU/L (303 for screening and 202 for monitoring thyroxine therapy) had an additional third-generation TSH analysis. With a second-generation assay for screening, 11% more free thyroxine (FT4) measurements were required to exclude thyrotoxicosis but there was a 42% saving on the reagent budget compared with a third-generation assay. In patients taking thyroxine, 33% more FT4 measurements were required to exclude over-replacement but the calculated saving in reagent costs was 53%. The costs of all other aspects of the two methods were similar. In this community-based sample, the improvement in sensitivity yielded by the third-generation assay at the lower end of the normal range reduced the number of confirmatory FT4 levels required to exclude thyrotoxicosis or over-replacement with thyroxine, but reagent costs were nevertheless higher than for second-generation assays. In financial terms, there is little justification for use of assays with sensitivity greater than the second generation (0.1 mU/L).

Biomarkers↗
PubMed1994

An outbreak of leptospirosis on a single farm in east Otago.

AIMS: To present an outbreak of leptospirosis on a dairy farm which had an unvaccinated herd. METHODS: Nine people working or living on the farm were tested for the presence of leptospira antibody by the microscopic agglutination test. Nineteen cows randomly selected, were also tested. RESULTS: Three human cases and one suspected case were detected over a one month period (December 1992-January 1993). Seventy-nine percent of the dairy herd tested had serological evidence of infection with L hardjo. CONCLUSIONS: Leptospirosis continues to be a major occupational problem in New Zealand. The importance of herd vaccination and the use of protective clothing during milking is emphasised.

Adolescent↗
PubMed1994

Comparison of goat and horse blood as culture medium supplements for isolation and identification of Haemophilus influenzae and Streptococcus pneumoniae from upper respiratory tract secretions.

The results of this study show that goat blood as a culture medium supplement is as supportive as horse blood for the isolation and identification of Haemophilus influenzae and Streptococcus pneumoniae from clinical material. Care is required in the preparation of goat blood chocolate agar to ensure that a thermolabile growth inhibitor of NAD-dependent Haemophilus species is eliminated.

Animals↗
PubMed1994

Upper airway carriage by Haemophilus influenzae and Streptococcus pneumoniae in Australian aboriginal children hospitalised with acute lower respiratory infection.

When nasopharyngeal secretions from 171 Australian Aboriginal children hospitalized with acute lower respiratory tract infections (ALRI) were cultured selectively for Streptococcus pneumoniae and Haemophilus influenzae, 136 (79.5%) and 151 (88.3%) children yielded 166 and 254 isolates of S. pneumoniae and H. influenzae, respectively. In colonized subjects multiple populations of S. pneumoniae (20% of carriage-positive patients) and H. influenzae (55%) were common. Pneumococci belonging to 27 types or groups were identified. H. influenzae serotype b colonized 16.4% of all children studied. More than one half of 152 children tested were excreting antibiotics at the time of admission to hospital. Significantly fewer children with serum antibiotic residues were colonized with S. pneumoniae than were antibiotic free children. Antibiotic usage had no measurable impact on the isolation rate of H. influenzae.

Acute Disease↗
PubMed1993

An outbreak of serotype 1 Streptococcus pneumoniae infection in central Australia.

An outbreak of serotype 1 Streptococcus pneumoniae infection involving both adults and children occurred in central Australia during the winter months of 1991. Eighteen patients, mainly Aboriginal men, presented with culture-positive serotype 1 bacteraemic pneumonia. In this group, 11 of 12 adults for whom medical records were available were alcohol dependent. Thirteen children who were separately studied were hospitalised with acute lower respiratory tract infection: none had bacteraemia but all had upper airway colonisation by type 1 pneumococci. Antibiotics taken by 8 of the 13 children before admission to hospital may have compromised the isolation of type 1 pneumococci from blood cultures. Since the availability of antibiotics, epidemic pneumococcal infection is infrequent and has not been reported in Australia. In three outbreaks of type 1 disease recorded elsewhere crowding and alcoholism were identified as contributory factors. In the 16 month period before this outbreak none of 162 strains of pneumococci isolated from blood (32 strains) and nasopharyngeal secretions (130 strains) from Aborigines with acute lower respiratory tract infection and meningitis in the Alice Springs region were serotype 1 organisms.

Adult↗
PubMed1993

Interferon-alpha (IFN alpha) induces a cytolytic mechanism in ovarian carcinoma cells through a protein kinase C-dependent pathway.

We have recently shown that IFN alpha induces a cytolytic mechanism in human ovarian carcinoma cell lines which is revealed when protein synthesis is subsequently inhibited. In order to determine whether the cytolytic activity induced by IFN alpha was activated through a pathway involving the activation of PKC, the human ovarian carcinoma cell line Caov-3 was exposed to phorbol 12-myristate 13-acetate (PMA). Under conditions where PMA activates PKC, PMA mimicked IFN alpha in its ability to induce a cytolytic mechanism. In contrast, under conditions where PMA depletes PKC, PMA not only did not induce cytolytic activity, but it prevented IFN alpha from inducing cytolytic activity. To further investigate the involvement of PKC in the signaling of cytolytic activity by IFN alpha, the ability of the protein kinase inhibitors, staurosporine, 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine dihydrochloride (H7), N-[2-(methylamino) ethyl]-5-isoquinolinesulfonamide dihydrochloride (H8), and N-(2-guanidinoethyl)-5-isoquinolinesulfonamide dihydrochloride (HA1004) to block the induction of cytolytic activity by IFN alpha was determined. The fact that H7, H8, and staurosporine, but not HA1004, blocked the induction of cytolytic activity by IFN alpha provides additional evidence of the involvement of PKC in this activity. Taken together these results indicate that the cytolytic activity induced by IFN alpha is induced through apathway that involves the activation of PKC.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗