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Biomedical subjects

G R Davis

Publications and source records attributed to G R Davis.

At least 19 recordsLinked to original sources

Characterization of growth hormone releasing factor analog expression in Saccharomyces cerevisiae.

An analog of growth hormone releasing factor (GRF), [Leu27]GRF(1-40)-OH, has been expressed and secreted in Saccharomyces cerevisiae under the control of the alpha-factor gene promoter and prepro sequence. A single pair of consecutive basic residues served as a processing site between the alpha-factor sequences and the GRF sequences. [Leu27]GRF(1-40)-OH from fermentor broth containing 20-30 mg/L of immunoreactive peptides was shown to be correctly processed and to possess biological activity as measured in vitro and in vivo. Additional peptides purified from broth appear to result from proteolytic degradation of the original translation product. Analysis of the amino acid compositions and sequences of these peptides suggests that processing enzymes may be responsible for some of the degradation.

Amino Acid Sequence

Mauthner cells maintain their lumbar projection in adult frog.

We have used retrograde labeling with horseradish peroxidase (HRP) in the bullfrog. Rana catesbeiana, to determine whether Mauthner (M) cells maintain a projection to the lumbar spinal cord in adult bullfrogs. We found that M cells persist in the adult bullfrog and maintain a projection to the lumbar spinal cord, despite the degeneration of much of their afferent input and of their motoneuronal targets in the spinal cord.

Animals

Detection of human immunodeficiency virus type 1 in AIDS patients using amplification-mediated hybridization analyses: reproducibility and quantitative limitations.

Eighty-six peripheral blood mononuclear cell (PBMC) samples from 30 patients with AIDS were analyzed using a transcription-based amplification system (TAS) and the polymerase chain reaction (PCR). Human immunodeficiency virus type 1 (HIV-1) sequences were detected by amplification-mediated hybridization in 98% of the samples, 52% of which were positive for p24 antigen by ELISA. Neither TAS (93%) nor PCR (95%) detected HIV-1 sequences in all 86 samples. The hybridization-detection methods (slot blot, bead-based sandwich, and solution) used to detect the HIV-1-specific TAS products had a clear influence on the efficiency of detecting and quantitating the levels of HIV-1 present in these samples. The reproducibility of amplification of constant amounts of HIV-1 RNA and beta-globin DNA by TAS and PCR was studied over 3 months. The results indicated that variations of 10- and 5-fold in the HIV-1 sequence levels could be detected between samples by TAS and PCR, respectively. Within the range of sensitivities for each assay used, the administration of zidovudine did not appear to reduce the amount of HIV-1 nucleic acid sequences as observed in PBMC obtained serially from six AIDS patients.

Acquired Immunodeficiency Syndrome

Transcription-based amplification system and detection of amplified human immunodeficiency virus type 1 with a bead-based sandwich hybridization format.

The in vitro amplification of biologically important nucleic acids has proceeded principally by a strategy of DNA replication. Polymerase chain reaction was the first such protocol to achieve this goal. In this report, a transcription-based amplification system (TAS) is described. Each cycle of the TAS is composed of two steps. The first is a cDNA synthesis step that produces one copy of a double-stranded DNA template for each copy of RNA or DNA target nucleic acid. During the course of this cDNA synthesis step, a sequence recognized by a DNA-dependent RNA polymerase is inserted into the cDNA copy of the target sequence to be amplified. The second step is the amplification of the target sequence by the transcription of the cDNA template into multiple copies of RNA. This procedure has been applied to the detection of human immunodeficiency virus type 1 (HIV-1)-infected cells. After four cycles of TAS, the amplification of the vif region of the HIV-1 RNA genome was measured to be, on the average, 38- to 47-fold per cycle, resulting in a 2-5 x 10(6)-fold increase in the copy number of the original target sequence. This amplification by the TAS protocol allows the detection of fewer than one HIV-1-infected CEM cell in a population of 10(6) uninfected CEM cells. Detection of the TAS-generated RNA from HIV-1-infected cells can easily be accomplished by means of a bead-based sandwich hybridization protocol, which provides additional specificity for the identification of the amplified HIV-1-specific sequence.

Cell Line

The double collision technique: a new method for measurement of the motor nerve refractory period distribution in man.

A new, double collision, technique is described for non-invasive measurement of the motor nerve refractory period distribution for human peripheral nerves. In contrast to previous collision techniques, this method is independent of the transient changes in nerve and muscle fibre conduction which can distort test muscle responses. The end-point of the distribution is determined by a null response; this permits accurate identification of those nerve fibres with the longest refractory periods. We have used the double collision technique to measure the refractory period distributions for the median nerve at the wrist in 20 normal subjects aged between 22 and 58 (mean 35) years. 150% maximal stimuli were used with a controlled limb temperature of 35 degrees C. Following a conditioning stimulus, the mean latencies for recovery of 5%, 50%, 95% and 99% of motor nerve fibres were 0.94, 1.03, 1.12 and 1.23 msec respectively. Data are also presented for the ulnar and peroneal nerves. The results show that the human motor nerve refractory period distribution is much less dispersed than has been previously supposed.

Adult

Motor nerve conduction velocity distributions in man: results of a new computer-based collision technique.

A new computer-based collision technique for direct measurement of the human motor nerve conduction velocity distribution is described. In contrast to previous collision techniques, the test muscle response is progressively cancelled to a null using an arrangement of proximal and distal stimuli which eliminates distortion of the test response caused by transient changes in nerve and muscle fibre conduction. The increased sensitivity of this new technique permits accurate measurement of the slowest 1% of alpha motor nerve fibres. We have used our modified collision technique to determine motor nerve conduction velocity distributions for the median nerve in 20 normal subjects aged between 19 and 59 (mean 35) years. 150% maximal stimulus intensities were used, with a controlled limb temperature of 35 degrees C. Group mean velocities (+/- S.D.) for the fastest (95%), mean (50%) and slowest (5% and 1%) motor fibres were 59.1 +/- 3.0, 56.9 +/- 2.9, 52.7 +/- 3.1 and 51.2 +/- 3.7 m/sec respectively. Data are also presented for the ulnar and peroneal nerves.

Action Potentials

The effect of continuous voluntary activation on neuromuscular transmission: a SFEMG study of myasthenia gravis and anterior horn cell disorders.

In normal subjects the neuromuscular jitter does not increase during continuous voluntary activation. In patients with myasthenia gravis, spinal muscular atrophy and motor neurone disease we have found that the neuromuscular jitter may increase during recordings of several minutes of continuous voluntary activation at steady innervation rates. In some units this led to impulse blocking, and in other units an initially normal jitter increased beyond the normal range. Measurement of jitter during continuous voluntary activation at steady innervation rates provides relevant information in the evaluation of neuromuscular transmission and fatigue in these disorders.

Adult

Feeding responses by adult males of Melanoplus sanguinipes (Fabricius) to carbohydrates and amino acids.

Adult males of Melanoplus sanguinipes (Fabricius), the migratory grasshopper, were starved for 18 h and were placed, one per arena, into arenas containing treated glass-fibre discs. The discs had been dipped into aqueous solutions of carbohydrates or amino acids of 0.0001, 0.001, 0.01, or 0.1 M concentration and allowed to dry at air temperature. They were placed vertically on pins and distributed at random among five equidistant stations around the periphery of the bottom of the arena. Each arena was provided with a glass-fibre disc that had been dipped in distilled water, as a control. The grasshoppers were allowed to feed at room temperature at 16:8 (L:D) for 24 h. Determinations of the effects of nutrient chemicals on feeding were determined by dry-weight differences between the weights of the treated discs before and after testing. Ribose, glucose, mannose and trehalose had no influence on feeding; 0.01 M fructose of 0.1 M arabinose, galactose, melibiose or sucrose increased feeding over that obtained with the control discs (P = 0.05); and 0.01 M xylose or sorbose depressed feeding. Each of the 11 amino acids investigated was phagodeterrent to adult males of M. sanguinipes. The present results confirm the variability of responses to carbohydrates among acridids; however, the reversal of phagodeterrency of sorbose or xylose at 0.1 M concentration is believed to be the first reported instance of such behaviour. In contrast to other reports of phagostimulation of acridids by individual amino acids or amino-acid mixtures, all protein amino acids tested in the present study were phagodeterrent to adult males of M. sanguinipes, even at low concentrations.

Amino Acids

Effects of amino-acid mixtures on food utilization and growth in Tenebrio molitor L.

Ten holidic diets, varying in amino-acid concentration or composition, were fed to larvae of Tenebrio molitor for four weeks at 27 +/- 0.25 degrees C and 65 +/- 5% r.h. Effects of diet on growth, food utilization and energy utilization were recorded for individual larvae. Differences in gains in fresh weight or in dry matter among larvae fed diets containing 0% to 5% of the amino-acid mixture were not demonstrated. However, larvae fed 10% or 20% of this mixture gained more than the former, but less than larvae fed a diet of ground whole wheat and brewer's yeast (9:1, w/w). When the amino-acid mixture was supplemented with alanine, aspartic acid, and serine, or with these three and asparagine, and was fed to larvae at the 10% level, growth was slower than with the unsupplemented mixture. Supplementation of the amino-acid mixture with the first three amino acids did not reduce larval growth when fed at the 20% level. Amounts of food and of energy utilized were positively correlated with larval fresh-weight and dry-matter gains. Energy utilization was negatively correlated with dietary amino-acid level. Food and energy utilization and larval growth were influenced by dietary amino acids, either metabolically or through phagostimulation.

Amino Acids

Somatostatinoma syndrome. Biochemical, morphologic and clinical features.

Diabetes mellitus, steatorrhea, cholelithiasis and a tumor distorting the duodenum prompted a work-up for somatostatinoma in a 52-year-old man. The responses of pancreatic B-cells but not of A-cells to nutrient stimuli were inhibited, and growth-hormone release was suppressed, suggesting somatostatin resistance in some target tissues. Plasma somatostatin-like immunoreactivity ranged from 9000 to 13,000 pg per milliliter (normal: 88+/-8, mean +/- S.E.M.) and was distributed in four molecular forms, including free somatostatin. The primary tumor contained 5 microgram of somatostatin-like immunoreactivity per milligram of wet tissue, distributed in three of the molecular forms noted in plasma. Plasma calcitonin was also elevated (4650 pg per milliliter; normal: less than 120). Immunocytochemical studies showed that cells of the primary tumor contained somatostatin and calcitonin but no other peptide hormones. Only somatostatin was present in the metastases. Somatostatin was localized electron microscopically in all secretory granules, irrespective of size and shape, whereas calcitonin was present only within a single subpopulation of small granules in the same cells.

C-Peptide

Protein nutrition of Tenebrio molitor L. XX. Growth response of larvae to graded levels of amino acids.

Larvae of Tenebrio molitor L;, Gembloux strain, race F, having an average initial weight of 10 mg were reared for 4 weeks at 27 +/- 0.25 degrees C and 65 +/- 5% relative humidity on diets containing an amino-acid mixture resembling the composition of larval tissues; Each of 14 amino acids was tested individually at levels of 0, 10, 20, 50, 100 and 200% of the amount found in larval tissues, while the concentration of other amino acids remained constant. Diets were maintained isonitrogenous at 1.6% nitrogen with supplementary glutamic acid, glycine, serine and tyrosine; Maximum fresh weight gains and dry weights were achieved by larvae fed reference levels of all essential amino acids except phenylalanine, threonine and tryptophan. Maximum growth was achieved by larvae fed 50% of the phenylalanine level found in larval tissues, Threonine and tryptophan were the limiting amino acids in this study and are probably required in the diet in excess of twice the concentration occuring in larval tissues, Probable ranges for quantitative amino-acid requirements of T. molitor were determined and suggestions were made for improving the nutritional adequacy of the amino-acid mixture.

Amino Acids

Ileal chloride secretion as a cause of secretory diarrhea in a patient with primary intestinal tuberculosis.

Severe secretory diarrhea resulting in dehydration and hypokalemia was observed in a patient in whom primary cecal tuberculosis was diagnosed by colonoscopy. Although the ileum appeared normal at colonoscopy and by biopsy, intestinal perfusion studies showed that the ileum was secreting water and electrolytes. Chloride secretion against a high electrical gradient suggested active anion secretion. Absorption in the colon was unimpaired. The diarrhea resolved after 4 wk of antituberculous therapy. Repeat perfusion studies after 3 mo showed normal absorption in the ileum. This case report suggests that intestinal tuberculosis can cause secretory diarrhea. The mechanism by which secretion occurs is not clear, but probably involves active anion secretion.

Cecal Diseases

Fecal incontinence in chronic diarrhea. Report of a case with improvement after training with rectally infused saline.

Patients with chronic diarrhea and fecal in continence are unable to retain as much rectally infused saline as patients without incontinence. We explored the effect of training such a patient to retain rectally infused saline. The patient was a 31-yr-old female with chronic diarrhea of obscure etiology who had daily episodes of fecal incontinence which markedly restricted her lifestyle. Training was accomplished by urging the patient to retain as much of a 25-min rectal infusion of 1500 ml saline as possible. After 10 training sessions, the patient increased her ability to hold rectally infused saline almost sevenfold. This increase was well maintained over 10 wk. In spite of continued diarrhea, the patient's incontinence did not recur after the first week of training, and she was able to resume a normal life. Anal sphincter pressure and a test of continence for a solid sphere did not change during or after training. This simple training technique has potential as a treatment for disabling fecal incontinence in patients with chronic diarrhea.

Adult

Determination of useful barley selections in an improvement program for increased lysine content by larvae of Tenebrio molitor L.

Larvae of the yellow mealworm, Tenebrio molitor L., Gembloux strain, race F, were reared for 4 weeks at 27 +/- 0.25 degrees C and 65 +/- 5% relative humidity. They were fed on each of 22 cultivars of barley, at the protein level occurring in harvested seed and at a protein level of 10% of dietary protein. Growth and body composition of the larvae were correlated positively and significantly with the concentrations of basic amino acids in the barleys and negatively and significantly with the concentrations of leucine. The percentage of crude protein in larval tissues can be used as a measure of available lysine in barley cultivars, and gains in fresh weight of larvae as indices of arginine concentrations. Differences were evident between the biological and chemical estimations of these amino acids. Several of the Saskatoon barley selections, derived from crosses with Hiproly, were equal to Hiproly or Risø varieties in the amounts of lysine available to the larvae.

Amino Acids

Nutritional evaluation of wheat and barley cultivars by growth rate and body composition of larvae of Tenebrio molitor.

Larvae of the yellow mealworm, Tenebrio molitor L., Gembloux strain, race F, were reared on diets of 17 cultivars of wheat and 29 cultivars of barley, prepared for determination of digestible energy with mice, for 4 weeks at 27 +/- 0.25 degrees C and 65 +/- 5% relative humidity. Values for percentage crude protein of tissues of larvae fed wheat cultivars were significantly and positively correlated with values for digestible energy as determined with mice. These values were not correlated for larvae fed barley cultivars; however, values for per cent dry matter content of larvae were significantly and positively correlated with values for digestible energy determined with mice. This apparent discrepancy is explained on the basis of the chemical constitution of barley and the availability of amino acids of barley to the larvae. Use of larvae of Tenebrio molitor to indicate the digestible energy of cereal grains is feasible, provided that the proper parameter is chosen. Nevertheless, use of this biological method seems more suitable for evaluation of protein quality and of amino acid availability than for a measure of digestible energy of feeds.

Animal Nutritional Physiological Phenomena