Search PubMed⌕ Search

Biomedical subjects

M L Johnson

Publications and source records attributed to M L Johnson.

At least 37 records · Page 2Linked to original sources
PubMed2000

Background suppression in frequency-domain fluorometry.

Gated detection is often used in time-domain measurements of long-lived fluorophores for suppression of interfering short-lived autofluorescence. However, no direct method has been available for gated detection and background suppression when using frequency-domain fluorometry. We describe a direct method for real-time suppression of autofluorescence in frequency-domain fluorometry. The method uses a gated detector and the sample is excited by a pulsed train. The detector is gated on following each excitation pulse after a suitable time delay for decay of the prompt autofluorescence. Under the same experimental conditions a detectable reference signal is obtained by using a long lifetime standard with a known decay time. Because the sample and reference signals are measured under identical excitation, gating and instrumental conditions, the data can be analyzed as usual for frequency-domain data without further processing. We show by simulations that this method can be used to resolve single and multiexponential decays in the presence of short lifetime autofluorescence.

Fluorescence↗
PubMed2000

The effects of dietary available phosphorus levels and phytase on performance of young and older laying hens.

In the first of two experiments (20 to 70 wk of age), eight treatments consisted of corn-soybean meal diets (0.34% total P, 3.8% Ca, 17% CP, 2,758 kcal ME/kg) containing 0.10, 0.15, 0.20, 0.25, or 0.45% available P (AP), with the three lowest AP diets supplemented with 300 units of phytase/kg of diet. A second experiment evaluated the effect of feeding the 0.10% AP diet with and without phytase on performance and time required for onset of P deficiency in older hens (70 to 76 wk of age). In Experiment 1, the 0.10% AP diet with no supplemental phytase depressed performance by 28 wk of age. No other significant differences in performance were observed among treatments for the entire 20- to 70-wk period except that the 0.15% AP diet with no phytase resulted in body weights and tibia ash that were lower (P < 0.05) than those of hens fed the 0.45% AP diet during the last 30 wk. The mean daily AP intake of hens fed the 0.10% AP, 0.15% AP, 0.45% AP, or 0.10% AP + phytase diets was 94,159, 499, or 108 mg, respectively. Excreta P concentration was decreased by approximately 50% in birds consuming 0.10% AP + 300 U/kg phytase compared with those consuming 0.45% AP. In the second experiment, P deficiency signs occurred within 3 wk of consuming the unsupplemented corn-soybean meal diet (0.10% AP) compared with 8 wk in Experiment 1. The results of this study indicate that phytase improves P utilization in corn-soybean meal diets for laying hens and that a corn-soybean meal diet containing 0.15% AP (159 mg AP/d) or containing 0.10% AP + 300 units of phytase/kg (108 mg AP/d) supported optimal egg production from 20 to 70 wk of age. Additionally, results suggested that older hens may exhibit P deficiency symptoms sooner than younger hens.

6-Phytase↗
PubMed2000

Human immunodeficiency virus type 1 pathogenesis in SCID-hu mice correlates with syncytium-inducing phenotype and viral replication.

Human immunodeficiency virus type 1 (HIV-1) patient isolates and molecular clones were used to analyze the determinants responsible for human CD4(+) thymocyte depletion in SCID-hu mice. Non-syncytium-inducing, R5 or R3R5 HIV-1 isolates from asymptomatic infected people showed little or no human CD4(+) thymocyte depletion in SCID-hu mice, while syncytium-inducing (SI), R5X4 or R3R5X4 HIV-1 isolates from the same individuals, isolated just prior to the onset of AIDS, rapidly and efficiently eliminated CD4-bearing human thymocytes. We have mapped the ability of one SI HIV-1 isolate to eliminate CD4(+) human cells in SCID-hu mice to a region of the env gene including the three most amino-terminal variable regions (V1 to V3). We find that for all of the HIV-1 isolates that we studied, a nonlinear relationship exists between viral replication and the depletion of CD4(+) cells. This relationship can best be described mathematically with a Hill-type plot indicating that a threshold level of viral replication, at which cytopathic effects begin to be seen, exists for HIV-1 infection of thymus/liver grafts in SCID-hu mice. This threshold level is 1 copy of viral DNA for every 11 cells (95% confidence interval = 1 copy of HIV-1 per 67 cells to 1 copy per 4 cells). Furthermore, while SI viruses more frequently achieve this level of replication, replication above this threshold level correlates best with cytopathic effects in this model system. We used GHOST cells to map the coreceptor specificity and relative entry efficiency of these early- and late-stage patient isolates of HIV-1. Our studies show that coreceptor specificity and entry efficiency are critical determinants of HIV-1 pathogenesis in vivo.

Amino Acid Sequence↗
PubMed2000

Deconvolution analysis of cardiac natriuretic peptides during acute volume overload.

Cardiac natriuretic peptides, especially amino terminal pro-Brain Natriuretic Peptide (NT-proBNP), are emerging as powerful circulating markers of cardiac function. However, the in vivo secretion and elimination (t1/2) of these peptides during acute volume overload have not been studied. We present the first report of the secretion and elimination of cardiac natriuretic peptides, based on deconvolution analysis of endogenous ovine plasma levels measured by specific radioimmunoassay. Four normal, conscious sheep underwent rapid right ventricular pacing (225 bpm) for 1 hour to stimulate acute cardiac natriuretic peptide release. Plasma samples and right atrial pressure measurements were taken at regular intervals 30 minutes before, during, and 4 hours after pacing. Baseline right atrial pressure significantly increased (P:=0.02) during the 1 hour of pacing in association with a prompt increase in plasma BNP (P:=0.03), atrial natriuretic peptide (P:=0.01), and NT-proBNP (P:=0.02). Deconvolution analysis showed that the t1/2 of NT-proBNP (69.6+/-10.8 minutes) was 15-fold longer than BNP (4.8+/-1. 0 minutes). Despite sustained increases in atrial pressure, cardiac secretion of natriuretic peptides (particularly atrial natriuretic peptide) fell during the pacing period, suggesting a finite source of peptide for secretion. Size-exclusion high-performance liquid chromatography revealed NT-proBNP to be a single immunoreactive peak, whereas BNP comprised at least 2 immunoreactive forms. These findings, especially the prompt secretion of BNP and the prolonged t1/2 of NT-proBNP, clarify the metabolism of BNP forms and help to explain the diagnostic value of NT-proBNP measurement as a sensitive marker of ventricular function.

Algorithms↗
PubMed2000

Hangover symptoms in Asian Americans with variations in the aldehyde dehydrogenase (ALDH2) gene.

OBJECTIVE: Hangovers are not experienced by all people and whether they contribute to the development of alcoholism is unclear. One population that might provide some insight into the role of hangover in the etiology of alcohol use disorders is that of individuals of Asian heritage. Certain Asians have lower rates of alcohol use and alcoholism, findings associated with a mutation in the aldehyde dehydrogenase (ALDH2) gene. Asians with ALDH2*2 alleles drink less and are less likely to be alcoholic than Asians without this mutation. Following alcohol ingestion, they exhibit more intense reactions to alcohol and generate higher levels of the metabolite acetaldehyde. This study evaluated hangover symptoms in Asian Americans with variations in the ALDH2 gene. METHOD: Men and women of Chinese, Japanese and Korean heritage (N = 140) were asked about their drinking history and a blood sample was collected for genotyping at the ALDH2 locus. Subjects used a Likert-type scale to estimate their severity of hangover and completed a 13-item hangover scale assessing the frequency of hangover symptoms during the previous 6 months. RESULTS: With abstainers (n = 17) excluded and with the effects of gender and recent drinking history controlled, ALDH2 genotype accounted for a significant amount of additional variability in the estimated severity of hangover score with a similar, but nonsignificant, trend for a five-item subscale score derived from the hangover scale. CONCLUSIONS: These results suggest that Asian Americans with ALDH2*2 alleles may experience more severe hangovers that may contribute, in part, to protection against the development of excessive or problematic drinking in this population.

Adult↗
PubMed2000

Fragment size difference between multiplex and singleplex PCR products and their practical implications.

By simultaneously amplifying several loci in the same reaction, multiplex PCR has been used in gene mapping and DNA typing with polymorphic short tandem repeat loci. Previous studies have discussed in detail the various parameters and conditions that influence the quantity of individual products generated by multiplex PCR. In practice, when a primer pair fails to amplify in a multiplex PCR for some individuals, singleplex PCR is often employed as a supplement to amplify the primer pair. However, the reliability of this procedure is unknown. In this study, we used six primer pairs from ABI PRISM Linkage Mapping Set version 2 to perform multiplex and singleplex reactions. The fluorescence-labeled amplification products were separated and detected on ABI PRISM 310 Genetic Analyzer. We found that for the marker D1S468, multiplex and singleplex reactions for the majority of individuals yielded reactions of different sizes. Therefore, the potential size difference between multiplex and singleplex reactions needs to be investigated. This investigation is essential to employ multiplex PCR supplemented with singleplex PCR in gene mapping and DNA typing.

DNA↗
PubMed2000

Relationship of dorsoventral eyeshine distributions to habitat depth and animal size in mesopelagic decapods.

Eyeshine distribution patterns recorded from the eyes of 19 mesopelagic decapod species were examined and related to the depths at which the species are found. For most species examined, eyeshine was found to be brighter ventrally than dorsally. Deep-water decapod species that do not undergo diel vertical migrations had brighter dorsal eyeshine than migratory species. Eyeshine intensity increased with body size in five of the species examined and decreased in two. These changes in eyeshine intensity may be an adaptation to variations in depth distributions that occur with increasing body size. It is suggested that the depth and size-related changes reflect the importance of remaining camouflaged in the mesopelagic realm and are an example of ecologically functional development.

Animals↗
PubMed1999

Association of bone dimensions with a parathyroid hormone gene polymorphism in women.

We have recently shown that bone radiogrammetric dimensions are associated with vitamin D receptor gene polymorphism. Since parathyroid hormone (PTH) plays a central role in maintaining calcium homeostasis and in bone remodeling, we investigated whether bone radiogrammetric dimensions are associated with a PTH gene polymorphism in 91 healthy Caucasian women, who were premenopausal at entry into the study. These women had assessments of bone by radiogrammetry every five years for a median period of 20 years (range 4-27). DNA was extracted from white blood cells. A segment of the PTH gene with a polymorphism at a BstBI restriction site was amplified by polymerase chain reaction. Diameter, cortical thickness and cross-sectional area at standard sites of the metacarpals, radius and femur were measured with radiogrammetry. Higher metacarpal diameter and cross-sectional cortical area, and a slower decrease in radial cortical area with age, were associated with the absence of the BstBI restriction site of the PTH gene. PTH gene polymorphism accounts for about 7-9% of the total variances of bone dimensional variables. These findings suggest that the dimensions of long bones are influenced by allelic variations in the PTH gene or in genes nearby.

Adult↗