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Biomedical subjects

L A Johnson

Publications and source records attributed to L A Johnson.

At least 109 records · Page 6Linked to original sources

Gender preselection in humans? Flow cytometric separation of X and Y spermatozoa for the prevention of X-linked diseases.

Human X- and Y-chromosome-bearing spermatozoa were separated based on their DNA content, using modified flow cytometric cell sorting technology. The resulting separation purity of the X-bearing from Y-bearing spermatozoa was evaluated using in-situ hybridization with alpha satellite DNA probes for the X- and Y-chromosomes. In the putative X-enriched-sorted populations, an average of 82% of the spermatozoa showed a hybridization signal with the X probe. Similarly, in the Y-sorted population 75% gave a signal with the Y probe. Sorted X- and Y-bearing spermatozoa were found to maintain their viability for several hours after sorting. These results demonstrate that the human sperm sex ratio can be significantly shifted to favour the selection of female-producing (X) spermatozoa or male-producing (Y) spermatozoa when spermatozoa are flow cytometrically sorted on the basis of DNA content. We propose that flow cytometrically sorted human spermatozoa, used in conjunction with in-vitro fertilization or intra-oviductal insemination, could be used by families who are at risk for X-linked diseases to preferentially produce female offspring. Sorted spermatozoa could also be used to pre-select for male offspring if that were medically indicated.

Cell Separation↗

Magnetic resonance imaging of overall and regional body fat, estrogen metabolism, and ovulation of athletes compared to controls.

The association of menstrual dysfunction of athletes with changes in body composition has been controversial, because most estimations of body fatness have been indirect. Using magnetic resonance imaging, we quantified the sc and internal fat over a specific volume from the fifth thoracic vertebra to femoral fat in the upper thigh and at 4 other anatomical landmarks of 17 athletes (13 oarswomen and 4 runners) compared to that in 11 nonathletic controls. The magnetic resonance imaging data were also analyzed for the athletes and controls in relation to ovulatory status, which was determined by assay of urinary pregnanediol glucuronide, and in relation to the extent of 2-hydroxylation of estradiol to a nonpotent metabolite, 2-hydroxyestrone, which was evaluated by radiometric analysis. We found that 1) the relative and absolute body fat values of the athletes were significantly less (P < 0.05) than those of the controls overall and at each of the six regional sites, although the body weights of the rowers were significantly heavier than those of the controls, and the runners did not differ from the controls; 2) the ratio of sc fat to internal fat was 80%:20% among both athletes and controls, even though the athletes had significantly less fat; 3) the extent of estradiol 2-hydroxylation was significantly (P = 0.005) inversely related to total fat as a percentage of the total volume and to sc fat as a percentage of the total volume (P = 0.004) overall and at each of the regional fat depots; 4) athletes with menstrual disorders had significantly decreased sc and internal fat overall and at all regional sites compared to controls; and 5) a subgroup of ovulatory rowers had an apparent increase or lack of decrease in internal fat at the level of vertebrae lumbar 4, sacral 1, and sacral 4, compared to controls, whereas their sc fat was decreased at these sites compared to that in controls. Changes in regional fat deposits of both sc and internal fat may be involved in the menstrual dysfunction of the athletes in addition to their decreased overall fatness. The body weight and body mass index of well trained athletes can be a misleading index of body composition.

Adipose Tissue↗

Experimental murine chronic hepatitis: results following intrahepatic inoculation of human uveitis mycoplasma-like organisms.

Mycoplasma-like organisms (MLO) are non-cultivated intracellular cell-wall deficient pathogenic bacteria with a distinctive ultrastructural appearance. Diagnosis of MLO disease depends on finding the organisms in parasitized cells using a transmission electron microscope. MLO are a well studied cause of transmissible chronic plant disease responsive to antibiotics. MLO have recently been found to cause human chronic uveitis, orbital, and retinal disease with autoimmune features. Ophthalmic leucocytes in these patients display MLO parasitization. Inoculation of human uveitis MLO into mouse eyelids produced chronic uveitis. MLO also disseminated to produce randomly distributed lethal systemic disease including chronic hepatitis. MLO parasitized leucocytes were present in all disease sites. Direct intrahepatic inoculation of human hepatic pathogens is a simple and efficient technique to produce murine hepatitis. This report describes the delayed onset widespread inflammatory liver disease produced by direct intrahepatic inoculation of human chronic uveitis MLO in 12 of 20 mice versus 0 in 40 controls (P < 0.05). The liver disease was accompanied by elevated serum SGOT levels, splenomegaly, and accelerated mortality. All 12 inflamed livers displayed MLO parasitized leucocytes versus 0 of 10 control livers. The resemblance of human chronic active hepatitis, massive hepatic necrosis, and post-necrotic cirrhosis to the MLO induced murine liver disease, the role of molecular biologic techniques in the detection and classification of those bacteria, and in therapy of MLO disease are discussed.

Animals↗

Analysis of variance of parameter estimates: F tests and t tests.

The problem of comparing and pooling experimentally independent estimates of a parameter such as a Michaelis constant (K) has been treated as a simple analysis of variance of "within" and "between" set deviations from the fitted variable (v). As applied to assessing the reproducibility of multiple estimates of the same K, this is identical to the procedure of Duggleby (Anal. Biochem. 189, 84-87, 1990). However, the theory developed here shows that applying Duggleby's procedure to the comparison of two experiments (each consisting of multiple data sets) depends critically on the assumption of equal errors within and between the individual sets, i.e., Fvb vw = s2wv/s2bv is close to 1. Application of the method when this is not the case will underestimate the common error (s2rv), overestimate its associated degrees of freedom (vr = vb+vw), and may suggest apparently significant differences where there are none. The theory also shows that this situation is an instance of the Fisher-Behrens problem and shows how Welch's solution can be applied. This gives the between set error s2bv as the corrected estimate of the common error and the corrected degrees of freedom as a simple function of vb, vw, and Fvb vw. When the nine prephenate dehydratase data sets which originally showed three apparently significant differences were reanalyzed in this way, all the variations in K were found to be within the range of the experimental error.

Analysis of Variance↗

Iodine content of commercially-prepared cat foods.

Twenty-eight varieties of commercially-available cat food (23 canned, 5 dried) were analysed for iodine. The iodine concentration varied from less than 0.37 micromol/kg to 41.8 pmol/kg, wet weight (less than 1.48 micromol/kg to 167 micromol/kg, dry weight). Excessive or insufficient iodine intake or wide swings in iodine intake over prolonged periods may contribute to thyroid disorders in cats.

Journal Article↗

Serum free thyroxine levels respond inversely to changes in levels of dietary iodine in the domestic cat.

Because of a perceived increase in the incidence of toxic multinodular goitres in cats in recent years, we investigated the iodine content of three varieties of commercial canned cat foods and studied the acute effects of 'ingestion of these preparations on urinary iodine excretion and serum free thyroxine levels in young, healthy cats. Ten castrated male cats were fed from a common source. The type of food was changed every 2 weeks. Urine and blood specimens were obtained weekly. Serum free thyroxine levels were determined and iodine concentrations in urine were assayed. The iodine content of the cats' food was also assayed. Food varieties of high, intermediate and low iodine content were fed for 2-week periods. There was a consistent, reciprocal relationship between the mean urinary iodine concentration and the mean serum free thyroxine level for each 2-week period. The difference in the mean serum free thyroxine concentrations for the high and low iodine intake periods was highly significant (p<0.01). When the serum free thyroxine level and the urinary iodine level for each cat at each collection throughout the 12-week study were analysed (66 paired results), a strong inverse correlation (r=0.59, p<0.01) was found. We concluded that the serum free thyroxine level in cats, as measured by a kit designed for human serum, is acutely responsive to changes in iodine intake.

Journal Article↗

Degradation of triglycerides by a pseudomonad isolated from milk: molecular analysis of a lipase-encoding gene and its expression in Escherichia coli.

Psychrotrophic lipolytic bacteria represent a significant problem in the storage of refrigerated dairy products. A lipase-encoding gene has been cloned and characterized from a highly lipolytic strain of Pseudomonas. The nucleotide sequence of the gene predicts a polypeptide of M(r) 49,905, which was identified when the gene was expressed in Escherichia coli.

Amino Acid Sequence↗

Quantitative chemical shift imaging of vertebral bone marrow in patients with Gaucher disease.

To evaluate extent of bone marrow involvement and disease severity in Gaucher patients, results of modified Dixon quantitative chemical shift imaging (QCSI) of the lumbar spine were correlated with quantitative analysis of marrow triglycerides and glucocerebrosides and with quantitative determination of splenic volume at magnetic resonance (MR) imaging. High-field-strength MR spectra of surgical marrow specimens were dominated by a single fat and a water peak, validating use of QCSI. QCSI showed average vertebral marrow fat fractions of 10% +/- 8 in Gaucher patients (normal adult averages, 29% +/- 6). Relaxation times for lipid and water approximated normal averages; bulk T1 values were significantly longer, reflecting decreased marrow fat. Glucocerebroside concentrations were higher in Gaucher marrow and inversely correlated with triglyceride concentrations. Extent of marrow infiltration determined by fat fraction measurements correlated with disease severity measured by splenic enlargement. These results show that as Gaucher cells infiltrate bone marrow and displace normal marrow adipocytes, bulk T1 increases due to the higher T1 of water compared with that of fat. QCSI provides a sensitive, noninvasive technique for evaluating bone marrow involvement in Gaucher disease.

Body Water↗

Multiple tumor suppressor genes in multistep carcinogenesis.

One of the most exciting areas of molecular oncology is the convergence of two independent lines of evidence suggesting involvement of multiple tumor suppressor genes in a given type of cancer. First, epidemiology and somatic cell genetics indicate the presence of multiple tumor suppressor genes in each of several malignancies. Second, cancers often lose multiple chromosomal regions during tumor progression. We will use two tumors, colorectal cancer and Wilms tumor, to illustrate the questions that multiple tumor suppressor genes raise.

Colorectal Neoplasms↗

A computer management system for patient simulations.

A series of interactive videodisc patient simulations is being used to teach clinical problem-solving skills, including diagnosis and management, to dental students. This series is called Oral Disease Simulations for Diagnosis and Management (ODSDM). A computer management system has been developed in response to the following needs. First, the sequence in which students perform simulations is critical. Second, maintaining records of completed simulations and student performance on each simulation is a time-consuming task for faculty. Third, the simulations require ongoing evaluation to ensure high quality instruction. The primary objective of the management system is to ensure that each student masters diagnosis. Mastery must be obtained at a specific level before advancing to the next level. The management system does this by individualizing the sequence of the simulations to adapt to the needs of each student. The management system generates reports which provide information about students or the simulations. Student reports contain demographic and performance information. System reports include information about individual patient simulations and act as a quality control mechanism for the simulations.

Computer Simulation↗

Iodine intake and the seasonal incidence of thyrotoxicosis in New Zealand.

We explored a possible relationship between the seasonal variation in the incidence of thyrotoxicosis in our area and iodine intake, as assessed by measuring the 24-h urinary iodide excretion rate eight times at 3-month intervals in a group of normal subjects and in 992 outpatient specimens over a 2-year period (1988-1990). For the period 1978-1990, 139 cases of thyrotoxicosis were diagnosed during the warmer half of the year (November-April) and 100 cases during the cooler half (May-October). This difference was statistically highly significant (P = 0.013). A similar disproportionality in the incidence of thyrotoxicosis was observed between the warmer (57%) and cooler (43%) halves of the year for the period 1988-1990. During the 1988-1990 period there was no discernible seasonal variation in either the milk iodide concentration or in the cumulative mean 24-h urinary iodide excretion rates for the normal subjects or the outpatients. We conclude that the higher proportion of thyrotoxic patients diagnosed during the warmer 6-month period of the year in our area is best explained by the fact that symptoms tend to be less tolerable in warm weather. A comparison of our findings on 24-h urinary iodide excretion rates with data collected in our area 25 years ago suggested that iodine intake may have declined.

Adult↗

A survey of the iodide concentration of human milk.

The iodide concentration of human milk from mothers living in the Wellington area and of six prepared infant formulae was measured. A total of 93 human milk samples were assayed, 14 from mothers of infants delivered at less than 35 weeks gestation and collected within 30 days of delivery (group A), 57 samples from mothers of infants delivered at 35 or more weeks gestation and collected within 60 days of delivery (group B) and 22 samples from mothers with infants aged greater than 60 days (group C). There was no significant (p greater than 0.05) difference between the means (SE) of groups A 1.14 mumol/L (0.20) and B 0.98 mumol/L (0.07). The mean for group C 0.39 mumol/L (0.03) was significantly less (p less than 0.05) than the means for both groups A and B. For groups A, B and C combined, there was a correlation (r = 0.44, p less than 0.01) between the milk iodide concentration and the iodide/creatinine ratio of a maternal urine specimen obtained immediately before or after the milk specimen. Formula milk preparations based on cow's milk contained higher concentrations of iodide than human milk.

Creatinine↗