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Biomedical subjects

J E McIntosh

Publications and source records attributed to J E McIntosh.

At least 37 records · Page 2Linked to original sources
PubMed1986

The analysis of data from breast cancer estrogen and progesterone receptor assays: Scatchard plots are inferior to direct fitting by computer.

The advantages are illustrated of analyzing breast cancer estrogen and progesterone receptor assays by fitting a receptor-ligand binding model directly to experimental data, rather than using the common graphical transformation known as the 'Scatchard plot'. Analysis of laboratory experiments and the use of simulation show that while both methods give equivalent results, the direct approach has definite advantages. In particular, results are more reliable in the face of increasing experimental error, it is possible to quantify non-specific binding from the primary data rather than having to measure it separately, and the methodology readily lends itself to optimizing the design of experiments.

Breast Neoplasms↗
PubMed1986

Immunohistochemical localization of calbindin-D28k during the development of the rabbit nephron.

Specific antibodies raised against a 28-kilodalton chick intestinal calcium-binding protein (calbindin-D28k) were used to localize the protein immunocytochemically in the developing rabbit kidney. Kidneys taken from rabbits between the 13th embryonic and 17th postnatal day were examined. Calbindin-D28k was observed in the mesonephric duct and the ureteral bud on the 13th embryonic day. During subsequent involution of the mesonephros, the ampullae of the metanephric ureteral buds contained calbindin-D28k. The protein was gradually lost from the ureters and the deep interstitial collecting ducts. Calbindin-D28k was never present in the renal vesicles derived from the nephrogenic blastema, but it was present in the connecting tubule segments during formation of the arcades. The last finding supports the belief that the connecting tubule is derived from the ureteral bud.

Animals↗
PubMed1986

Varying the patterns and concentrations of gonadotrophin-releasing hormone stimulation does not alter the ratio of LH and FSH released from perifused sheep pituitary cells.

Our aim was to determine whether release of LH and FSH can be controlled differentially by the characteristics of applied signals of stimulatory gonadotrophin-releasing hormone (GnRH) alone, free of the effects of steroid feedback or other influences from the whole animal. The outputs of both gonadotrophins were significantly correlated (r approximately 0.90; P less than 0.0005) when samples of freshly dispersed sheep pituitary cells were perifused in columns for 7 h with medium containing a range of concentrations of GnRH in various patterns of pulses. Hormone released in response to the second, third and fourth pulses from every column was analysed in detail. Dose-response relationships for both LH and FSH were very similar when cells were stimulated with 5-8500 pmol GnRH/l in 5-min pulses every hour. When GnRH was delivered in pulses at a maximally stimulating level, the outputs of both hormones increased similarly with increasing inter-pulse intervals. Efficiency of stimulation (release of gonadotrophin/unit stimulatory GnRH) decreased (was desensitized) with increasing pulse duration in the same way for both hormones. Thus, varying the dose, interval and duration of GnRH pulses did not alter the proportions of LH and FSH released in the short-term from freshly dissociated cells. However, the same cell preparations released more LH relative to FSH when treated with maximally stimulating levels of GnRH for 3 h in the presence of 10% serum from a sheep in the follicular phase of its ovulatory cycle compared with charcoal-treated serum.(ABSTRACT TRUNCATED AT 250 WORDS)

Anestrus↗
PubMed1986

Effects of lithium and phorbol esters on the dynamics of LH release from dispersed sheep pituitary cells.

The possible involvement of polyphosphoinositides in the stimulation of LH release was investigated. Dispersed sheep pituitary cells were incubated in test-tubes, or perifused in columns, with gonadotrophin-releasing hormone (GnRH) and Li+, or with a phorbol ester, and the amounts and patterns of LH release over time compared. Treatment with Li+ (10 mmol/l), which is known to increase levels of inositol phosphates in gonadotrophs, was shown to have effects only on the responses of desensitized cells, significantly decreasing the rate at which the cells desensitize (P less than 0.005) and decreasing the response to supramaximal levels of GnRH stimulus (P less than 0.01). It is suggested that these effects could be due to increased levels of inositol monophosphate, inositol bisphosphate or inositol 1,3,4-trisphosphate. Responses to single or repeated pulses of GnRH at 18-, 30- and 60-min intervals were not significantly altered. Phorbol 12-myristate 13-acetate (PMA), an activator of the calcium and phospholipid-dependent protein kinase (protein kinase C), was specifically active in releasing LH with a half-maximal stimulating dose of approximately 3 nmol/l. Phorbol 12,13-diacetate, which is structurally similar to PMA but does not activate protein kinase C, did not release LH, except at high levels in freshly dispersed cells. The timing of PMA-stimulated LH release was similar to that for GnRH-stimulated release, and PMA was able to release greater amounts of LH than could GnRH. This suggests that activation of protein kinase C is likely to be important in the GnRH-stimulated release of LH from gonadotrophs.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

The wheat mitochondrial gene for apocytochrome b: absence of a prokaryotic ribosome binding site.

The wheat mitochondrial gene for apocytochrome b (CYB) has been identified by its hybridization to a yeast CYB probe and its nucleotide sequence has been determined. The wheat CYB sequence predicts a cytochrome b apoprotein of 398 amino acids; it is almost identical to that of maize but has ten additional amino acids at the carboxy terminus. No introns are present in the wheat CYB gene, but an internal segment of the gene is repeated at another genomic location. Transcript analysis reveals a single wheat CYB mRNA of approximately 2.4 kb with a long untranslated leader. Sequences upstream of the CYB coding region are very similar in wheat and maize but the stretch proposed to be a ribosome binding site in maize is not conserved in wheat. The corresponding leader regions of the wheat mitochondrial mRNAs for cytochrome oxidase subunits I and II also lack complementarity to the 3'-end of the small subunit rRNA. We conclude that alternative signals are involved in the initiation of translation in plant mitochondria.

Amino Acid Sequence↗
PubMed1985

Dynamic characteristics of luteinizing hormone release from perifused sheep anterior pituitary cells stimulated by combined pulsatile and continuous gonadotropin-releasing hormone.

We aim to quantify and relate the dominant dynamic factors of GnRH signals and of the resultant patterns of LH release from pituitary cells. Using perifused sheep cells we have already shown that rising edges of GnRH pulses are major effectors of LH release and that a longer absence of signal between pulses improves response. This study reports the effects on LH release dynamics of continuous levels of GnRH with superimposed pulses and of slowing the important rising edge of the GnRH pulses. Low baseline GnRH perifusions at physiological levels (5-60 pM) reduced the response to hourly pulses of 850 pM GnRH. Continuous GnRH (420 pM), which initially yielded maximal LH release followed by desensitization, prevented extra stimulation by pulses of equal concentration, but 10-fold higher pulses gave additional LH output. After desensitization an hour's respite from stimulation resensitized cells to 420 pM pulses. Whereas continuous stimulation of cells with GnRH even at the very low level of 5-10 pM [ED50 = 58 +/- 6 (SE) pM] produced desensitization in 10-15 min, slowly rising GnRH (0.56-14 pM/min) caused increasing LH output with time. However, in comparison with square wave pulses, stimulatory signals consisting of slowly rising concentrations of GnRH produced peaks characterized by less total LH output and a changed shape. This was consistent with desensitization at low concentrations of GnRH reducing response to later increases in the level of stimulation. The mechanism for detecting GnRH signals and/or the mechanisms controlling release of LH were desensitized to constant GnRH at any concentration but retained a reduced sensitivity, or developed an additional release capacity, to increased levels of GnRH. Properties of four distinct types of LH release dynamics were described quantitatively and were shown to be controlled by different time constants in the GnRH pulse stimulation patterns.

Animals↗
PubMed1985

The effect of various corticotropin-releasing factor trains on the release of adrenocorticotropin, beta-endorphin, and beta-lipotropin from perifused ovine pituitary cells.

The dynamics of the release of proopiomelanocortin-derived hormones from ovine anterior pituitary cells in response to varying pulse characteristics of ovine corticotropin-releasing factor (CRF) were investigated with an in vitro automated 15-column simultaneous perifusion system. Columns of cells were stimulated continuously or with trains of CRF pulses of varying pulse length (2-16 min), pulse period (20-160 min), and concentration, for 500 min. Ovine ACTH, beta-lipotropin, and beta-endorphin immunoreactivity were measured by unextracted RIA. Each pulse of CRF stimulated clearly defined and highly correlated (r greater than 0.9) pulses of the three pituitary hormones, suggesting similar mechanisms controlling release. In dose-response experiments, the minimum concentration of CRF in a 10-min pulse required to significantly raise the output of ACTH was 200 pM, and initial responses had not attained maximal levels with concentrations of CRF increased to 2 microM. Responses to pulsed CRF stimulation decreased with time with all stimulation patterns selected, although previously unstimulated control columns retained the initial capacity to respond. Multiple linear regression analysis showed that hormone output per pulse of CRF (43 nM) increased with increasing pulse period and pulse length. Output of pituitary hormones per unit of CRF applied decreased with pulse length but increased with pulse period. In summary, the responses of proopiomelanocortin derivatives were shown to be sensitive to abrupt increases in CRF, to reduce output under continued stimulation, and to have an inherent time lag before responding maximally to subsequent pulsed stimulation.

Adrenocorticotropic Hormone↗
PubMed1985

Amplitude of episodic release of LH as a measure of pituitary function analysed from the time-course of hormone levels in the blood: comparison of four menstrual cycles in an individual.

Pulse amplitude and frequency are often used to describe measurements of LH in blood. Such analyses are compatible with models of LH being released from the pituitary in episodes that are controlled by pulses of hypothalamic gonadotrophin-releasing hormone. The amplitudes of these secretory episodes as seen in blood are usually defined as the net heights of peaks above a baseline. As a measure of each pituitary secretory episode, this is valid only if peaks are regularly and widely spaced, making overlap negligible. When episodes are erratic and frequent so that only fractions of peaks have been cleared from the circulation before others follow, nadirs between peaks include output from previous episodes and do not define a physiologically meaningful baseline. Applied to overlapping peaks, such measures of amplitude usually underestimate pituitary secretory episodes and imply a tonic mode of LH secretion in addition to pulsatile release. Using the additional information of fitted LH clearance coefficients to define the shapes of LH peaks, a simple method based on an episodic mode of release alone is described, for estimating more accurately the relative sizes of secretory episodes as observed in blood, free of the effects of overlapping peaks. Using this analysis we have described the variation in amplitude, interval and clearance rates of LH secretory episodes within and between four normal menstrual cycles of a single individual. Thirteen, 3-6 h blood sampling sessions were performed during early follicular growth at the transition from luteal to follicular phases when the frequencies of LH peaks, LH/FSH ratios and progesterone concentrations were changing markedly.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1983

Radioimmunological measurements of total LH in sheep pituitary cells.

Procedures commonly used to extract LH from pituitary cells in order to measure total cell content were compared in four cell preparations. It was shown in 81 samples of cells suspended in 1 mM EDTA or 50 mM NaHCO3 that after freezing and thawing followed by any of a variety of treatments, there were no significant differences in the amounts of LH measured by RIA relative to the arbitrarily chosen reference treatment of vigorous pipetting. The additional treatments were multiple freezing and thawing, homogenisation, sonication, homogenisation in 25-100 mM Na2CO3 followed by rapid neutralisation, or none. The consistency of the results suggested that the same cellular pools of LH were being made accessible for measurement with all treatments. However, use of the more vigorous conditions of 1-2.5 M urea, 1% Triton X-100, or sonication on ice in 100 mM Na2CO3 decreased the amount of measurable hormone presumably due to its modification. In two cell preparations, homogenisation of cells in 100 mM Na2CO3 produced an additional 45% of measurable LH not accessible using other treatments nor from the source material in two other preparations.

Animals↗
PubMed1983

Influence of the characteristics of pulses of gonadotrophin releasing hormone on the dynamics of luteinizing hormone release from perifused sheep pituitary cells.

The effects were studied of varying the frequency, width and amplitude of pulses of gonadotrophin releasing hormone (GnRH) on the release of LH from anterior pituitary cells. Dispersed sheep cells supported in Sephadex were perifused with medium for 10 h and stimulated with different constant pulse patterns of GnRH. The timing of release of LH was measured by radioimmunoassay of the effluent fractions. Pulses of GnRH ranging in duration from 2 min every 8 min to 16 min every 128 min, and in concentration from 1.7 pmol/l to 250 nmol/l were applied to the cells, as well as continuous stimulation. Comparisons of differences between LH release patterns among samples of the same preparation of cells were used to demonstrate the effects of different GnRH stimulatory regimes. It was concluded that (1) the frequency of GnRH stimulation was important to the nature of LH release (periods shorter than about 16 min between pulses reduced LH output and caused faster desensitization of response), (2) the pulse width of GnRH input was important (the rising edge of the pulse produced greater LH output per unit of GnRH input than did continued application of GnRH within a pulse and wider pulses combined with shorter periods reduced LH output) and (3) over a threshold value of 5-10 nmol GnRH/l pulse amplitude had little further influence on LH output or rate of desensitization in dispersed cells. These findings reinforce the hypothesis that the rising edge of the GnRH pulse is the major stimulant to LH release.

Animals↗
PubMed1982

Immunocytochemical localization of vitamin D-dependent calcium-binding protein in renal tubules of rabbit, rat, and chick.

Vitamin D-dependent calcium-binding protein (CaBP) was localized in tissue sections of kidneys from rabbits, rats, and chicks using antiserum specific for chick intestinal CaBP. In rabbit kidney, CaBP was present in all cells of the distal convoluted tubule and most cells of the connecting tubule. Fewer, but still a majority, of the cells of cortical collecting ducts contained CaBP. The intensity of immunochemical staining and the number of stained cells decreased markedly in medullary collecting ducts, and only a few collecting duct cells contained CaBP at the junction of the inner and outer medulla. In the rat kidney, CaBP was present in all distal convoluted tubule cells, but the immunochemical staining was less intense than in the rabbit. The protein also was found in most connecting tubule cells of the rat; however, only a few collecting duct cells in the superficial corte of the rat contained CaBP. CaBP was essentially absent from mid- to deep-cortical collecting duct cells, while a very few collecting duct cells always contained CaBP at the junction of the inner and outer stripes of the outer medulla. In the chick, CaBP was present in distal convoluted tubule cells as the distal convoluted tubule coursed adjacent to the central vein. CaBP was absent from chick collecting duct cells. In all three species CaBP was not detected in the other portions of the nephron.

Animals↗
PubMed1981

Neonatal body water turnover: a putative index of perinatal morbidity.

The water metabolism of 46 newborn babies was determined during a 10 day period by means of an isotope dilution technique, and correlations were sought with the clinical assessment of the babies by multiple obstetric and pediatric clinical criteria. The babies, 48 to 72 hours of age, were given a single oral dose (2 ml/kg) of deuterated water (D2O), a nonradioactive tracer, and the urinary excretion rate was followed by means of infrared spectrophotometry. After a period of equilibration of the D2O with body water (20 hours), the rate of D2O clearance was found to be a single exponential decay process, thus allowing the fraction of total body water lost each hour (the rate constant) to be calculated for each baby. The median values of the rate constants X 10(4)(h-1) for 14 growth-retarded babies ws 104 (98% confidence limits, 97.8 to 122) compared with 76.3 (67.0 to 80.2) for 16 normal mature babies and 82.1 (73.4 to 90.6) for 16 normal premature babies. These data indicate that, compared with normal mature or normal premature babies, growth-retarded infants have a significantly (P less than 0.05) faster turnover of water during the first 10 days of postnatal life. Since there was little overlap in results between the normally grown and the retarded infants, the measurement of water turnover may provide a useful index of perinatal morbidity.

Body Water↗
PubMed1981

Urinary steroid metabolites and the overgrowth of lean and fat tissues in obese girls.

Studies were made of steroid metabolites excreted in the urine of 17 obese girls 11.4 to 16.8 yr and 17 normal girls 11 to 17 yr. Creatinine excretion (muscle mass), total body water (or deuterium space), lean body mass and body fat were determined in the obese girls. Extracellular volume (corrected bromide space) was also measured and by difference with body water, intracellular water or soft tissue cell mass was calculated. In normal girls 24-h creatinine excretion was determined, but body water was predicted from height and weight. It was found, as in previous studies, that the obese girls had excess muscle mass and soft tissue cell mass for height. The excess growth of muscle, lean tissue, and body length in obese girls correlated with increments in oxosteroid (17 ketosteroid) excretion. The overall weight increase correlated with increased excretion of corticosteroid metabolites--a finding of interest since a physiological Cushing's syndrome was postulated for fat girls many years ago. When the normal and obese girls were divided by age at 14 yr and the subgroups compared (normal obese) the younger girls showed differences with respect to height, weight, total body water, fat and percentage fat. Differences in steroid metabolites were not found. In older girls the same findings were made again, but here it was clear that the increments in body size, particularly muscle mass, correlated with augmented oxosteroid excretion. Evidence is cited that these findings are not just related to a larger steroid pool in obese girls.

17-Ketosteroids↗
PubMed1981

Pulpal reaction to bleaching of teeth in dogs.

Hydrogen peroxide and heat are two components of a technique commonly used to bleach human teeth. The effects of these two components on pulp tissue of dog teeth were evaluated histologically. Hydrogen peroxide alone or with heat caused obliteration of odontoblasts, hemorrhage, resorption, and inflammatory infiltration, while heat alone was not detrimental. Pulpal changes demonstrated evidence of reversibility after 60 d.

Animals↗
PubMed1981

The role of the pineal gland in seasonality.

The life time reproductive performance of 2 flocks of Merino crossbred ewes pinealectomized at 7-60 days of age, and maintained in South Australia did not differ from that of sham-operated control animals kept in the same flocks. The pineal gland is therefore not a major determinant of reproductive success, but a role for the pineal in adjusting breeding activity to season is not excluded. It was confirmed that pineal denervation of adult ewes by cranial sympathectomy could have disruptive effects and evidence of a seasonal change in pineal function was obtained from studies of plasma levels of melatonin, a putative pineal hormone, and a pineal peptide with gonadotrophin releasing activity. That melatonin stems mainly, but not exclusively, from the pineal gland was confirmed by studies of urinary excretion values. Adult Merino ewes produced 39-90 micrograms melatonin/day, mostly (80%) during the dark period, whereas pinealectomized animals produced less than one-fifth this amount. A link was provided between the pineal gland and diet. Melatonin and other 5-methoxyindoles influenced the activity of alpha-chymotrypsin and may regulate the availability of precursor substances required for the synthesis of neurotransmitters important in reproduction.

Animals↗
PubMed1980

Predicting the luteinizing hormone surge: relationship between the duration of the follicular and luteal phases and the length of the human menstrual cycle.

Determination of blood serum levels of luteinizing hormone (LH) are used to detect the day of the midcycle surge. This information, collected over several menstrual cycles of numerous women, is used to derive mathematical expressions relating the day of the surge to the length of the cycle. The equations are subsequently employed to predict the most likely day of the LH surge, and hence the time of ovulation, solely from knowledge of the average length and variability of a woman's cycles, without the need for determinations of LH. A convenient table is provided for making this prediction.

Adult↗