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Biomedical subjects

R Johnson

Publications and source records attributed to R Johnson.

At least 91 records · Page 5Linked to original sources
PubMed1999

Abnormal morphogenesis but intact IKK activation in mice lacking the IKKalpha subunit of IkappaB kinase.

The oligomeric IkappaB kinase (IKK) is composed of three polypeptides: IKKalpha and IKKbeta, the catalytic subunits, and IKKgamma, a regulatory subunit. IKKalpha and IKKbeta are similar in structure and thought to have similar function-phosphorylation of the IkappaB inhibitors in response to proinflammatory stimuli. Such phosphorylation leads to degradation of IkappaB and activation of nuclear factor kappaB transcription factors. The physiological function of these protein kinases was explored by analysis of IKKalpha-deficient mice. IKKalpha was not required for activation of IKK and degradation of IkappaB by proinflammatory stimuli. Instead, loss of IKKalpha interfered with multiple morphogenetic events, including limb and skeletal patterning and proliferation and differentiation of epidermal keratinocytes.

Abnormalities, Multiple↗
PubMed1999

Molecular karyotypes for Alternaria plant pathogens known to produce host-specific toxins.

There are at least ten plant diseases caused by Alternaria species in which host-specific toxins (HSTs) are responsible for fungal pathogenicity. Of these HST-producers, seven are considered distinct pathotypes of the species Alternaria alternata, and the remaining three are among other species of pathogenic Alternaria. Inter- and intra-specific variation among Alternaria taxa, including HST-producers, was determined by electrophoretic karyotyping using pulsed-field gel electrophoresis. A. alternata including seven pathotypes of A. alternata and eight non-pathogenic strains had 9-11 chromosomal bands with estimated sizes ranging from 0.4 to 5.7 Mb. In contrast, Alternaria species that are morphologically distinct from A. alternata had 8-10 bands with sizes between 0.9 and 5.7 Mb. Estimated genome sizes of A. alternata and other Alternaria species ranged from 28.8 to 33.6 Mb and 25.1 to 30.7 Mb, respectively. Other species of pathogenic Alternaria were difficult to differentiate from A. alternata on the basis of chromosome-size polymorphisms alone, but Southern analysis using rDNA as a probe could, in some cases, differentiate between them. These results were cytologically confirmed by 4',6-diamidino-2-phenylindole (DAPI) staining and fluorescence in situ hybridization with a rDNA probe for mitotic metaphase chromosomes prepared by the germ-tube burst method.

Alternaria↗
PubMed1999

In vitro synaptotrophic effects of Cerebrolysin in NT2N cells.

Recent studies have shown that Cerebrolysin can enhance synaptic function and ameliorate synaptodendritic alterations in animal models of neurodegeneration, suggesting a synaptotrophic effect. We hypothesize that Cerebrolysin might exert this effect, in part, by regulating the expression of amyloid precursor protein (APP). We studied the patterns of expression of synaptic proteins during differentiation of human teratocarcinoma cell line NTera 2 (NT2) in the presence or absence of Cerebrolysin. This study showed that the terminally differentiated neurons (NT2N) expressed a wide variety of synaptic markers and that expression of these synaptic-associated proteins coincided with the shift in expression from APP770/751 to APP695. Furthermore, APP immunoreactivity was colocalized with synaptophysin-immunoreactive neuritic varicosities in NT2N neurites, and Cerebrolysin treatment of NT2N cells resulted in an augmented and earlier expression of synaptic-associated proteins. This increased synaptic protein expression coincided with an increase in APP695 over APP770/751. These results support the possibility that synaptotrophic effects of Cerebrolysin might be mediated via regulation of APP expression.

Amino Acids↗
PubMed1999

The use of laser scanning cytometry to assess depth of penetration of adenovirus p53 gene therapy in human xenograft biopsies.

SCH58500 is an agent for gene therapy of cancer, consisting of a replication-deficient type 5 adenovirus (Ad5) expressing the human p53 tumor suppressor gene (Ad5/p53). An important question about the use of Ad5/p53 gene therapy is how to achieve the therapeutically effective delivery of an Ad5/p53 vector to the tumor. We wanted to determine the effective depth of penetration of an Ad5/p53 vector by dosing the vector in an experimental human xenograft/SCID model. To assess depth of penetration, we developed a novel methodology for scanning tissue sections by laser scanning cytometry (LSC). SCID mice were given intraperitoneal injections of either p53(null) SK-OV-3 human ovarian tumor cells or p53(mut) DU-145 human prostate tumor cells to establish xenograft solid tumors. Mice were then dosed once or twice at 24-hour intervals by intraperitoneal injection with SCH58500 (Ad5/p53), an adenovirus construct expressing beta-galactosidase (Ad5/beta-gal), or a buffer control. Additional groups of mice received a single intraperitoneal dose of 10 mg/kg paclitaxel either alone or coadministered with Ad5/p53. Twenty-four hours after each last dose, the human solid tumor xenograft and relevant mouse tissue were removed from each mouse for the analysis of Ad5/p53 penetration. Immunohistochemistry (IHC) for beta-galactosidase protein revealed a depth of penetration of between 1 and 10 cells from the tumor surface. In some mice, hepatocytes in the periportal regions of liver lobules were also positive, indicating systemic absorption of adenovirus from the peritoneal cavity. IHC staining for p53 and p21 proteins in SK-OV-3 solid tumor xenografts revealed similar Ad/p53 penetration. LSC was used to map and quantitate apoptosis in both tumor and liver tissue biopsies, with over 450,000 nuclei from liver tissue and 150,000 nuclei from tumor tissue being evaluated. LSC analysis demonstrated a high level of apoptosis in the tumors that had been removed from Ad5/p53-dosed mice (12.7-19.7%). This level of apoptosis was significantly higher (P < 0.05) than was observed for liver tissues taken from Ad5/p53-dosed mice (2.7-8.0%) or tumor tissues taken from either Ad5/beta-gal-dosed mice (3.0-6.4%) or buffer control-dosed mice (3.0-5.3%). Scan bit maps from the extensive LSC analyses confirmed that apoptosis was present to about the same depth (1-10 cells) as had been identified by IHC for beta-galactosidase, p53, and p21 proteins. Paclitaxel coadministered with Ad5/p53 had no effect on Ad5 penetration into solid tumors in vivo as measured by IHC for p53 or p21 protein. However, the combination therapy did cause an elevation in the number of tumor cells undergoing apoptosis.

Adenoviridae↗
PubMed1999

Soluble dietary fiber protects against cholesterol gallstone formation.

BACKGROUND: Epidemiological studies have suggested that soluble dietary fibers are hypocholesterolemic and may inhibit cholelithiasis. METHODS: Thirty prairie dogs were placed on a cholesterol-supplemented lithogenic diet. Ten animals received 5% psyllium (PSY) and 10 animals received 5% cellulose. After 6 weeks all gallbladders were inspected for stones; blood and bile were collected for analysis. RESULTS: Cholesterol stones were present in 8 of 10 of the control animals, in 6 of 10 of the cellulose group, and 3 of 10 of the PSY animals (P <0.05). Concentrations of cholesterol and chenodeoxycholic acid (CDCA) were significantly lower in the PSY group compared with controls (0.49 versus 0.88 mM and 4.2 versus 9.2 mM, respectively) leading to a significant reduction in the cholesterol saturation index (0.62 versus 1.2). CONCLUSIONS: A dietary soluble fiber (PSY) inhibits cholesterol stone formation by reducing the biliary cholesterol saturation index. This protective effect is associated with a selective decrease in biliary cholesterol and CDCA.

Animals↗
PubMed1999

Platelets and DAMI megakaryocytes possess beta-secretase-like activity.

We report here the discovery of two novel human platelet and megakaryocytic DAMI cell enzymes that have beta-secretase-like activity. These activities could potentially effect cleavage of the amyloid precursor protein (APP) at the beta-amyloid peptide N-terminus, by an EC 3.4.24.15-like metalloprotease, and the N terminus-1 position, by a serine protease. Thus both enzymes may generate the amyloidogenic beta-peptide. Studies of intact and Triton X-100-lysed DAMI cells, as well as intact versus subcellular fractions of platelets, demonstrate the presence of these proteolytic activities. The resting platelet has (1) a surface serine protease, demonstrated by its ability to cleave a beta-secretase substrate and by its inhibitor sensitivity; and (2) a metalloprotease, recognized by an antibody to EC 3.4.24.15, which resides intracellularly in the alpha-granule membrane, is translocated to the surface on activation, and shows beta-secretase-like activity by cleaving the same substrate. This metalloprotease can also cleave recombinant APP to a potentially amyloidogenic fragment. Surface metalloprotease was identified in DAMI cells by flow cytometry and Western blotting with a specific anti-EC 3.4.24.15 monoclonal antibody, while activity was identified by using two beta-secretase substrates. This article is the first to document two previously unknown endoproteinases with beta-secretase-like activity in platelets and DAMI cells. These proteases are capable of effecting cleavage of APP and could therefore contribute to Abeta deposition in the cerebrovasculature.

Alzheimer Disease↗
PubMed1999

Helicopter transport of patients to tertiary care centers after cardiac arrest.

Air transport is commonly used to transfer survivors of cardiac arrest from rural hospitals to large tertiary-care centers, presumably to improve outcome. To examine this issue, a retrospective review of patients stabilized after a cardiac arrest was conducted; 157 transports were reviewed. The mean age of patients was 37.9 +/- 27.8 yrs, with a male to female ratio of 2.2:1. Survivors were significantly older than nonsurvivors. Thirty-one of 69 patients (45%) with primary cardiac disease were discharged alive from the hospital, 75% without neurological sequelae. Only a minority of patients with noncardiac medical illness (7%), electrical injury (33%), suffocation (15%), near-drowning (15%), and inhalation (0%) were discharged alive from the hospital. Outcomes for cardiac arrest in adult patients older than 65 years (32.3% survival) were similar to those for adult patients younger than 65 years (36.2% survival) (P = .887). These results show that survivors of a primary cardiac event have a favorable outcome when transferred by air to tertiary centers when compared with historical controls that were transported by ground. On the other hand, cardiac arrests from noncardiac medical illness, suffocation, near-drowning, and inhalation have a grim prognosis. Prospective studies should clarify the role of air transport in these patients.

Adolescent↗
PubMed1999

Mutation inhibition by beta-estradiol after low doses of gamma-irradiation of mammalian cells.

The methodology previously described for measuring mutagenesis has been applied to the study of mutation prevention in immortalized G2 phase human lymphocytes exposed to 25 and 50 cGy of gamma-radiation. Caffeine prevents repair of mutations. Two times 10(-4) M beta-estradiol applied for 2.5 h markedly decreases induced mutations and affects male and female cells similarly. Quantitative measurement of mutagenesis in cells of different individuals and the effect of various agents on mutation yield should be important in prevention of cancer and other mutational disease.

Caffeine↗
PubMed1999

Predicting body composition from anthropometry in pre-adolescent children.

The objectives of this paper were to: a) evaluate the accuracy and precision of previously published pediatric body composition prediction equations and b) develop additional prediction equations from a large, heterogeneous group of Caucasian (n = 133) and African-American (n = 69) children. The combined cohort of 202 children included a wide range of ages (4.0-10.9 y), weights (14.0-70.8 kg), fat mass (FM: 1.2-28.5 kg) and percent body fat (% body fat: 6.2-49.6%). Skinfold measurements were obtained using a Lange caliper and body fat was measured with a Lunar DPX-L densitometer. The previously published equations of Slaughter et al and Goran et al did not accurately predict body fat. The entire cohort was randomly divided into two sub-groups for purposes of deriving and cross-validating a new prediction equation. In stepwise regression analysis in the development group (n = 135), weight, triceps skinfold, gender, ethnicity and abdominal skinfold estimated FM measured by dual energy x-ray absorptiometry (DEXA) with a model R2 of 0.95. The new prediction equation was cross-validated in the control group (n = 67) and each ethnic and gender subgroup. We conclude that a) the equations of Slaughter et al and Goran et al did not accurately predict FM in a heterogeneous group of children and b) a new anthropometric prediction equation is proposed that may provide accurate estimates of FM in both Caucasian and African-American children aged 4-10 y with a wide range of FM and body composition.

Alabama↗
PubMed1999

Congenital orbital teratoma.

A case of congenital orbital teratoma is described in which there was no organized eye only microscopic evidence of ocular tissues within the disorganized teratoma. A baby boy presented at birth with a 10-x-8-cm mass extruding from the left orbit. Magnetic resonance imaging (MRI) showed a mixed cystic-solid orbital mass containing areas of calcification and deforming the bony orbit around its margins. There was no organized eye and no intracranial extension. The eye was removed with reconstruction of the eyelids. Histopathology showed representation from all three germ cell layers consistent with a teratoma. There was no organized eye, but some disorganized ocular structures within the teratoma. Follow-up has been uneventful. Neonatologists and pediatricians should be aware of the possible diagnoses in a newborn presenting with an orbital mass, so that early definitive surgery can be performed with preservation of the globe where possible.

Anophthalmos↗
PubMed1999

Influence of lower limb pneumatic compression on pulmonary artery temperature: effect on cardiac output measurements.

OBJECTIVES: To characterize the decreases in pulmonary artery temperature that coincide with the inflation cycle of pneumatic calf compression stockings and to examine their effects on the thermodilution measurement of cardiac output. DESIGN: Three-part observational study. SETTING: University hospital surgical intensive care unit. PATIENTS: Postoperative patients with indwelling pulmonary artery catheters. INTERVENTION: Thermodilution cardiac output measurements with and without pneumatic calf compression. MEASUREMENTS AND MAIN RESULTS: Phase 1 (n = 18) examined the effects of pneumatic compression on pulmonary artery temperature. There was no effect on pulmonary artery temperature (device off, 37.468+/-0.008 degrees C; device on, 37.458+/-0.014 degrees C), but the difference between the maximum and minimum pulmonary artery temperatures was increased (off, 0.031+/-0.006 degrees C; on, 0.055+/-0.012 degrees C [p < .001]). Phase 2 (n = 12) found that the mean thermodilution cardiac output with 10 mL of cold (0-5 degrees C) injectate was unchanged by pneumatic compression (off, 7.00+/-2.28 L/min; on, 6.89+/-2.22 L/min). However, when the compression devices were operating, the variability between the individual measurements was increased, as reflected by larger coefficients of variation (off, 3.19+/-1.96; on, 8.72+/-6.56 [p < .02]). Similar results were obtained during phase 3 (n = 5), when cardiac output was measured with room temperature Injectate. CONCLUSIONS: Intermittent pneumatic calf compression increased lower limb venous return, causing acute but transient decreases in pulmonary artery blood temperature. This did not affect the accuracy of thermodilution cardiac output measurements that were made using 10 mL of either cold or room temperature injectate.

Bandages↗
PubMed1999

Multiple layers of cooperativity regulate enhanceosome-responsive RNA polymerase II transcription complex assembly.

Two coordinate forms of transcriptional synergy mediate eukaryotic gene regulation: the greater-than-additive transcriptional response to multiple promoter-bound activators, and the sigmoidal response to increasing activator concentration. The mechanism underlying the sigmoidal response has not been elucidated but is almost certainly founded on the cooperative binding of activators and the general machinery to DNA. Here we explore that mechanism by using highly purified transcription factor preparations and a strong Epstein-Barr virus promoter, BHLF-1, regulated by the virally encoded activator ZEBRA. We demonstrate that two layers of cooperative binding govern transcription complex assembly. First, the architectural proteins HMG-1 and -2 mediate cooperative formation of an enhanceosome containing ZEBRA and cellular Sp1. This enhanceosome then recruits transcription factor IIA (TFIIA) and TFIID to the promoter to form the DA complex. The DA complex, however, stimulates assembly of the enhanceosome itself such that the entire reaction can occur in a highly concerted manner. The data reveal the importance of reciprocal cooperative interactions among activators and the general machinery in eukaryotic gene regulation.

Allosteric Regulation↗
PubMed1999

Caffeine overdose in a premature infant: clinical course and pharmacokinetics.

The elimination of caffeine was investigated in a 1860 g, 31 week gestation neonate, following the accidental administration of a 160 mg.kg-1 dose. The first serum concentration measured was 217.5 mg.l-1 at 36.5 h after dosing. Fitting of time-concentration data was performed using non-linear regression with MKMODEL. A first order elimination model was superior to a mixed order model. Parameter estimates were: clearance 0.01 l.h-1, volume of distribution 1.17 litres, elimination half-life 81 h. Toxic manifestations included hypertonia, sweating, tachycardia, cardiac failure, pulmonary oedema and metabolic disturbances (metabolic acidosis, hyperglycaemia and creatine kinase elevation). An unusual feature of this infant's illness course was gastric dilatation. These signs resolved by day 7 at a serum concentration of 60-70 mg.l-1. Caffeine clearance has traditionally been reported as either an absolute value or as directly proportional to body weight. The per kilogram model gives an erroneous impression that clearance is greatest in early childhood and then decreases with age until adult rates are reached in late adolescence. Age-related clearance values reported in the literature were reviewed using an allometric 3/4 power model. This size model demonstrates that clearance increases in infancy and reaches adult rates within the first three months of life.

Age Factors↗
PubMed1999

Factor structure and subtest differences on the Preschool Behavior Questionnaire in a Latino, African-American, Euro-American, and Asian preschool population.

This study examined the factor structure of the Preschool Behavior Questionnaire in a sample of African-American, Euro-American, Asian, and Hispanic children. The sample consisted of 304 children (141 boys, 163 girls) 3- and 4-yr.-old and enrolled in Head Start. A principal component analysis with a varimax rotation was conducted and two- and three-factor solutions were extracted. A two-factor solution produced a clear interpretive structure representing Fowler and Park's 1979 Aggressive-Hyperactive-Distractible and Anxious-Fearful factors. Even though a three-factor solution was statistically appropriate, extracting more than two factors yielded dimensions difficult to interpret. Examination of subscale differences among ethnic groups indicated significant group effect for ethnicity. Further examination showed that Euro-American children are rated significantly higher on the Anxious subtest than Latino, African-American, and Asian children, but there were no other subscale differences. Clinical and research implications are discussed.

Black or African American↗