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Biomedical subjects

K E Jones

Publications and source records attributed to K E Jones.

9 recordsLinked to original sources

A glass/silicon composite intracortical electrode array.

A new manufacturing technique has been developed for creating silicon-based, penetrating electrode arrays intended for implantation into cerebral cortex. The arrays consist of a 4.2 mm x 4.2 mm glass/silicon composite base, from which project 100 silicon needle-type electrodes in a 10 x 10 array. Each needle is approximately 1,500 microns long, 80 microns in diameter at the base, and tapers to a sharp point at the metalized tip. The technique used to manufacture these arrays differs from our previous method in that a glass dielectric, rather than a p-n-p junction, provides electrical isolation between the individual electrodes in the array. The new electrode arrays exhibit superior electrical properties to those described previously. We have measured interelectrode impedances of at least 10(13) omega, and interelectrode capacitances of approximately 50 fF for the new arrays. In this paper, we describe the manufacturing techniques used to create the arrays, focusing on the dielectric isolation technique, and discuss the electrical and mechanical characteristics of these arrays.

Electric Conductivity

Molecular cloning of a complementary deoxyribonucleic acid encoding the thyrotropin-releasing hormone receptor and regulation of its messenger ribonucleic acid in rat GH cells.

Rat pituitary GH cells have been used extensively to study the biochemical actions of TRH on lactotropic cells. To investigate the structure and regulation of the rat TRH receptor (rTRHR), we have cloned its cDNA from GH4C1 cells. Using the polymerase chain reaction with degenerate primers and pools of cloned cDNAs from a GH4C1 cDNA library, a fragment sharing high similarity to the mouse thyrotrope TRHR (mTRHR) was identified. Conventional library screening with this fragment was used to isolate a single cDNA. mRNA synthesized in vitro from this cDNA was injected into Xenopus oocytes, and a characteristic conductance response to TRH was detected by voltage clamp recording. DNA sequence analysis revealed a molecule of 412 amino acid residues, with 96% similarity to the mTRHR. However, in contrast to the mTRHR, the rTRHR had an additional 19 amino acid residues at its carboxy-terminus. A mRNA of about 4 kilobases was identified in GH3 cells. Regulation of the rTRHR mRNA concentration was studied in GH3 cells. Steady state rTRHR mRNA levels were decreased to 30% of the control level by incubation with TRH for 48 h and increased 4-fold by incubation with dexamethasone for 12 h. Southern blot analysis of genomic DNA from GH3 cells gave a simple banding pattern consistent with a single copy gene. We conclude that the rTRHR shares high primary sequence similarity to the mTRHR, but the rTRHR has an extension of 19 amino acids at its carboxy-terminus, which is lacking in the mTRHR.

Amino Acid Sequence

A silicon-based, three-dimensional neural interface: manufacturing processes for an intracortical electrode array.

A method has been developed for the manufacture of a "three-dimensional" electrode array geometry for chronic intracortical stimulation. This silicon based array consists of a 4.2 x 4.2 x 0.12 mm thick monocrystalline substrate, from which project 100 conductive, silicon needles sharpened to facilitate cortical penetration. Each needle is electrically isolated from the other needles, and is about 0.09 mm thick at its base and 1.5 mm long. The sharpened end of each needle is coated with platinum to facilitate charge transfer into neural tissue. The following manufacturing processes were used to create this array. 1) Thermomigration of 100 aluminum pads through an n-type silicon block. This creates trails of highly conductive p+ silicon isolated from each other by opposing pn junctions. 2) A combination of mechanical and chemical micromachining which creates individual penetrating needles of the p+ silicon trails. 3) Metal deposition to create active electrode areas and electrical contact pads. 4) Array encapsulation with polyimide. The geometrical, mechanical, and electrical properties of these arrays should make them well suited as interfaces to cortical tissue.

Animals

Deterioration in blood glucose control in females with diabetes changed to a basal-bolus regimen using a pen-injector.

Although the basal-bolus regimen of insulin delivery is more physiological than conventional twice daily insulin it does not usually improve blood glucose control. Having recently withdrawn 3 patients from pen-injector therapy for deteriorated blood glucose control we examined the records of all our diabetic patients on a pen-injector for at least 1 year. Of the 51 patients identified, 3 had died, 2 moved away, and 4 patients (3 females) had pen-injector therapy withdrawn for poor control; 3 patients had inadequate records. The remaining 21 males and 18 females were well matched for age, duration of diabetes, and duration of pen-injector treatment. Blood glucose control deteriorated significantly during pen-injector therapy in female patients (9.6 +/- 1.2 to 10.7 +/- 1.4%, p = 0.02), but not in the males (9.3 +/- 1.3 vs 9.4 +/- 1.4%, NS). Weight and insulin dose did not change significantly in any patient. Blood glucose control may deteriorate in young women with diabetes changed to insulin therapy with a pen-injector.

Adult

Differential regulation of thyroid hormone receptor messenger ribonucleic acid levels by thyrotropin-releasing hormone.

In addition to its well known actions in stimulating TSH and PRL synthesis and secretion, TRH has been shown to decrease the concentration of thyroid hormone receptors (TRs) in GH4C1 cells as measured by nuclear thyroid hormone (T3) binding. In the present study we have investigated the effects of TRH on the levels of mRNA encoding the different forms of TR, TR beta-1, TR beta-2, and TR alpha-1 as well as that of the non-T3-binding variant, c-erbA alpha-2. GH3 cells were incubated with 100 nM TRH in the presence or absence of 1 nM T3 for 48 h, and mRNA levels were determined by Northern blot analysis. Results revealed that there is differential regulation of the individual TRs by TRH at the pretranslational level. The mRNA for the pituitary-specific form of TR, TR beta-2, was down-regulated by 60% by TRH in GH3 cells, while that of its alternative splice product, TR beta-1, was unchanged. A modest change was observed in TR alpha-1 mRNA levels, which were down-regulated by 20%; there was no change in c-erbA alpha-2 mRNA levels. Levels of nuclear T3 binding were assessed under the same conditions, and 100 nM TRH was found to decrease binding by 40% from 0.78 to 0.46 fmol/micrograms DNA. A similar change in nuclear T3 binding was seen after incubation with 1 nM T3. The effect of TRH on the GH mRNA response to T3 was investigated. In the absence of TRH there was a 4-fold induction of GH mRNA after incubation with 1 nM T3. In the presence of 100 nM TRH, no significant induction in GH mRNA by T3 was seen, indicating that T3 responsiveness as well as receptor concentration are diminished by TRH under these conditions.

Adenoma

Triiodothyronine receptor beta-2 messenger ribonucleic acid expression by somatotropes and thyrotropes: effect of propylthiouracil-induced hypothyroidism in rats.

mRNA for a thyroid hormone receptor isoform that is unique to the pituitary gland (TR beta-2) is down-regulated by T3. Increases in the expression of this mRNA are seen in rats rendered hypothyroid by treatment with propylthiouracil (PTU). This study used dual labeling to determine which pituitary cells expressed TR beta-2 mRNA in normal and PTU-treated rats. In situ hybridization protocols localized the mRNA (with biotinylated complementary oligonucleotide probes detected by avidin-biotin-peroxidase), and immunoperoxidase protocols identified the pituitary hormone proteins. In dispersed pituitary cells, 20 +/- 2% (average +/- SD) of cells from normal rats and 30 +/- 3% of cells from PTU-treated rats were labeled for TR beta-2 mRNA. PTU caused increases in the area of the labeled cells (from 114 +/- 11 to 225 +/- 7 microns 2), the area of the label per cell (from 27 +/- 3 to 71 +/- 11 microns 2), and label density. PTU produced increases in the percentage of TSH cells from 8 +/- 1% to 19 +/- 2%, decreases in the percentage of GH cells from 27 +/- 3% to 11 +/- 2%, and no change in other cell types. After dual labeling, 73% of cells that expressed TR beta-2 mRNA stored either TSH (35 +/- 8) or GH (38 +/- 6). Less than 10% stored other hormones. When each cell type was analyzed, 56 +/- 3% of TSH cells and 43 +/- 4% of GH cells expressed TR beta-2 mRNA. When these percentages were multiplied by the percentages of each cell type in the overall population, TSH and GH cells with TR beta-2 mRNA represented 6.8 +/- 1% and 11.6 +/- 1% of the pituitary cells, respectively. Less than 1% of all pituitary cells expressed TR beta-2 and ACTH (0.9 +/- 0.06), LH (0.8 +/- 0.1), FSH (0.8 +/- 0.1), and PRL (0.9 +/- 0.04). PTU treatment increased the percentage of TSH cells with TR beta-2 mRNA to 72 +/- 4% and decreased the percentage of GH cells with TR beta-2 mRNA to 30 +/- 3%. However, some enlarged putative TSH cells could not be identified by immunolabel because the storage levels were low. Thus, changes in TR beta-2 mRNA in hypothyroid rats may be the net result of the increase in the percentage of TSH cells, the amount of mRNA per cell (measured by area and density of label), and the decrease in the percentage of GH cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

Isolation of a cDNA encoding human Rev-ErbA alpha: transcription from the noncoding DNA strand of a thyroid hormone receptor gene results in a related protein that does not bind thyroid hormone.

A cDNA encoding Rev-ErbA alpha, a member of the thyroid/steroid hormone receptor superfamily, has been isolated from a human fetal skeletal muscle library. This cDNA contains 269 consecutive base pairs identical to a region of a human c-erbA alpha-2 cDNA, but the respective long open reading frames utilize this nucleotide sequence in opposite orientations. Thus, human Rev-ErbA alpha (hRev) is derived from opposite-strand transcription of the c-erbA alpha genomic locus. mRNA encoding hRev was detected in human skeletal muscle by Northern analysis. Comparison of hRev and Rev-ErbA (rRev) reveals 99% identity in the putative DNA-binding and "ligand-binding" (carboxy-terminal) domains. hRev does not bind thyroid hormone (T3), as has also been found for its rat homolog. Interestingly, the human and rat Rev-ErbA alpha cDNAs are dissimilar at their 5' ends, corresponding to the first exon that we have identified in the rat gene. The conservation of the bidirectionally transcribed regions of c-erbA alpha-2 and Rev-ErbA alpha in human and rat suggests that this unusual genomic arrangement has an important function, perhaps related to the regulation of gene expression.

Base Sequence

Gerontological research. Is it useful for nursing practice?

The potential usefulness to gerontological nursing practice of nursing as well as non-nursing research was studied. The empirical phase of nursing research was also compared with that of medical, behavioral science, and multidisciplinary research. Although the quantity of nursing research reviewed was small, nearly 80% was judged to be potentially useful to the practice of gerontological nursing. About one third of the non-nursing research was judged potentially useful. These findings are relevant to all nurses, indicating the value on non-nursing as well as nursing research to nursing practice. Overall, the design of nursing research compared favorably to the design of non-nursing research.

Geriatric Nursing

A 100 electrode intracortical array: structural variability.

A technique has been developed for fabricating three dimensional "hair brush" electrode arrays from monocrystalline silicon blocks. Arrays consist of a square pattern of 100 penetrating electrodes, with 400 microns interelectrode spacing. Each electrode is 1.5mm in length and tapers from about 100 microns at its base to a sharp point at the tip. The tips of each electrode are coated with platinum and the entire structure, with the exception of the tips, is insulated with polyimide. Electrical connection to selected electrodes is made by wire bonding polyimide insulated 25 microns diameter gold lead wires to bonding pads on the rear surface of the array. As the geometrical characteristics of the electrodes in such an aray will influence their electrical properties (such as impedance, capacitance, spreading resistance in an electrolyte, etc.) it is desirable that such an array have minimal variability in geometry from electrode to electrode. A study was performed to determine the geometrical variability resulting from our micromachining techniques. Measurements of the diameter of each of the 100 electrodes were made at various planes above the silicon substrate of the array. For the array that was measured, the standard deviation of the diameters was approximately 9% of the mean diameter near the tip, 8% near the middle, and 6% near the base. We describe fabrication techniques which should further reduce these variabilities.

Animals