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Biomedical subjects

R S Turner

Publications and source records attributed to R S Turner.

At least 55 records · Page 3Linked to original sources

Activity of neurons in cerebellar-receiving and pallidal-receiving areas of the thalamus of the behaving monkey.

1. Thalamic neurons that receive synaptic input from the globus pallidus or the cerebellar nuclei were identified in awake monkeys trained to perform an arm-reaching task. The location of electrophysiologically identified cerebellar-receiving (CR) and pallidal-receiving (PR) neurons was used to identify a total of 264 thalamic neurons in cerebellar (CB) or pallidal (GP) regions of the thalamus. 2. Stimulation in the brachium conjunctivum or white matter adjacent to the cerebellar nuclei excited 85 neurons in the thalamus at short latencies. These CR neurons were located in the oral portion of the ventral posterolateral nucleus (VPLo), in caudal portions of the ventral lateral nucleus (VLc), and in area X. 3. Stimulation in the internal globus pallidus (GPi) inhibited 10 thalamic neurons at short latency. These PR neurons were located in rostral portions of VLc, in the oral part of the ventral lateral nucleus (VLo), and in the parvicellular part of the ventral anterior nucleus (VApc). 4. There was no clear single somatotopic organization of neurons in CB and GP regions of the thalamus, as defined by "free-form" responses to passive manipulation and observation of eye movements. There was, in fact, a tendency for two representations, each, of the head/eye/mouth cells and cells with modifications of activity in response to manipulation of the arm. 5. During the hold period before illumination of a visual target, the mean firing rates and variability of discharge of arm-related CR and PR neurons did not differ significantly. This was also true for the total sample of arm-related neurons in the CB versus GP regions. 6. The activity of many neurons in both the CB and GP regions began to change before the reaching movement and, for some, before the earliest recorded changes in electromyographic (EMG) activity. The initial change was an increase in discharge for greater than 75% of the cells studied in both the CB and GP regions. 7. During the reaching task, there also was no significant difference in the time of the initial change in discharge of neurons in the CB versus GP regions of the thalamus. 8. These data are consistent with the hypothesis that the initial task-related change in discharge of PR thalamic neurons is dominated by input from the cerebral cortex and that pallidal input modulates later phases of their movement-related changes in activity.

Animals↗

Synthetic myelin basic protein peptide analogs are specific inhibitors of phospholipid/calcium-dependent protein kinase (protein kinase C).

Synthetic peptide analogs of the bovine myelin basic protein (MBP) corresponding to residues 104-118 were found to specifically inhibit phospholipid/ Ca2+-dependent protein kinase (protein kinase C). The peptides [Ala107]MBP (104-118) and [Ala113]MBP (104-118) inhibited protein phosphorylation of intact MBP, histone H1 and peptide phosphorylation with MBP(104-123), MBP(104-118) or [Ala105]MBP (104-118) as substrates. The inhibitor peptides [Ala107]MBP(104-118) and [Ala113]MBP (104-118), containing alanine in place of the arginine recognition sites, apparently inhibited the enzyme noncompetitively with respect to substrates, with IC50 values ranging from 46-145 and 28-62 microM, respectively. These peptide analogs did not inhibit cyclic AMP-dependent protein kinase or myosin light chain kinase but inhibited phospholipid/Ca2+-dependent phosphorylation of endogenous proteins in the total, solubilized fraction of rat brain.

Amino Acid Sequence↗

Phospholipid/calcium-dependent protein kinase (protein kinase C) system: a major site of bioregulation.

The substrate specificity determinants of protein kinase C are probed using synthetic peptides encompassing the major phosphorylation site serine 115 in bovine MBP. The results indicate that basic residues arginine 107 and 113 N-terminal to the phosphorylation site are essential for the substrate activity of the peptides. Substitutions of these basic residues by alanine cause a marked decrease in their substrate activity and the resulting peptide analogs become specific and rather potent inhibitors of protein kinase C. Leukemic cells are particularly abundant in protein kinase C and its substrate proteins, and the enzyme system has been shown to play a key role in cell growth. The agents that stimulate protein kinase C include tumor promoting phorbol esters (such as TPA) and mezerein, and the putative second messenger diacylglycerol. Many antineoplastic agents, on the other hand, inhibit the enzyme which include adriamycin, tamoxifen, alkyl-lysophospholipid, selenium, retinal and lipoidal amine CP-46, 665-1. Immunocytochemical studies of protein kinase C in leukemic cells indicate that it is localized in the plasma membrane, cytoplasm, nucleus and Golgi apparatus, and the subcellular distribution of the enzyme might be related to the phases of the cell cycle. TPA induces translocation of the enzyme to plasma membrane, suggesting an additional mode of action for the tumor promotor.

Animals↗

Dynamic post-surgical hallux varus after lateral sesamoidectomy: treatment and prevention.

Hallux varus and clawing are occasionally seen after a McBride procedure, due to the severance of the flexor hallucis brevis, when the lateral sesamoid is removed. The present study includes six patients, (involving ten feet), who developed hallux varus and great toe clawing after McBride procedures were performed by various orthopedic surgeons. Treatment utilized partial proximal phalangeal resection, with and without silicone single-stem implants, extensor hallucis longus tendon transfer to the great toe metatarsal, and interphalangeal joint arthrodesis, or tenodesis of the great toe to correct clawing. One patient eventually required a great toe metatarsophalangeal joint fusion. This reconstructive surgery is generally effective by correcting capsular malalignment, the "bowstringing" tendency of the extensor hallucis longus, and great toe clawing that develops simultaneously with hallux varus. Lateral sesamoidectomy is risky, and believed unnecessary as adductor hallucis tenotomy is effective in relieving hallux valgus. Although, it can occur with excessive excision of the medial emminence and distal advancement of the abductor hallucis. The author is unaware of hallux varus developing after adductor hallucis tenotomies.

Adult↗

Substrate specificity of phospholipid/Ca2+-dependent protein kinase as probed with synthetic peptide fragments of the bovine myelin basic protein.

The substrate specificity of phospholipid/Ca2+-dependent protein kinase (protein kinase C) was studied using synthetic peptides, in particular those corresponding to the amino acid sequence around serine 115 in bovine myelin basic protein (MBP). It was found that MBP (104-118) and MBP (104-123) were substrates for the enzyme, with apparent Km values of 14 and 10 microM, respectively. Neither MBP (111-118) nor MBP (111-123) were phosphorylated, indicating that an additional segment of sequence extending toward the N terminus, but not toward the C terminus, was essential for the substrate activity of the peptides. Of the alanine-substituted analogs examined, [Ala 105] MBP (104-118) was comparable to the parent peptide, whereas [Ala 107] MBP (104-118) and [Ala 113] MBP-(104-118) were much poorer substrates. These findings indicated that lysine 105 was not essential, but both arginine 107 and arginine 113 were important specificity determinants. Initial studies revealed that [Ala 113] MBP (104-118) inhibited phosphorylation by the enzyme of the parent peptide and, to a lesser extent, the intact MBP(1-170). Serine 115 was the only site phosphorylated in the analog peptides [Ala 105] MBP (104-118) and [Ala 107]MBP (104-118). In the parent peptide, serine 115 was the initial site of phosphorylation but after prolonged phosphorylation other sites became phosphorylated (serine 110 and/or serine 112), further supporting the concept that arginine residues act as essential substrate specificity determinants for phospholipid/Ca2+-dependent protein kinase.

Animals↗

Afterimages of sinusoidal, square-wave and compound gratings.

Negative afterimages were observed after steady fixation of sinusoidal gratings at low spatial frequencies, and quantified using contrast-matching and cancellation methods. Afterimage contrast increased as a power function of "exposure", defined as the product of inspection contrast and inspection duration. A single function, linear on a log-log plot, describes the afterimages of gratings at different spatial frequencies, contrasts and durations. The matching method yielded a lower slope (about 0.42) than the cancellation method (0.62), probably because contrast adaptation attenuates perceived afterimage contrast in the first method, but not the second. Square-wave gratings, and those containing two or three harmonic components gave much weaker afterimages (assessed by contrast-matching) than sine-waves did. Contrast adaptation may again be responsible. Hemi-field asymmetries in contrast perception were noted. The contrast of real gratings and afterimages was nearly additive, especially at lower exposures, but the results cannot distinguish between linear filter and gain control models of "local adaptation".

Adaptation, Ocular↗

Correlation of the MMPI with lumbosacral spine fusion results. Prospective study.

This study was done to determine whether the Minnesota Multiphasic Personality Inventory (MMPI) aided in selection of appropriate lumbosacral fusion candidates, compared with those selected without an MMPI. Postoperative end-result ratings were done, categorizing patients into two groups: one group of 62 patients with a preoperative MMPI and a second group of 23 patients with no preoperative MMPI. Thirty patients with normal MMPIs, strong objective indications for initial surgery, and absence of compensation and/or litigation factors had 88% good or excellent end-results. Twenty-three patients had surgery without MMPI evaluation. Of 21 available for follow-up, 86% had good or excellent end-results. "Warning" physical signs aided selection of appropriate surgical candidates in the patients who did not have an MMPI.

Female↗

Phospholipid-sensitive Ca2+-dependent protein kinase inhibition by R-24571, a calmodulin antagonist.

R-24571 (calmidazolium), a derivative of the antimycotic agent miconazole, inhibited phospholipid-sensitive Ca2+-dependent protein kinase (PL-Ca-PK), with an IC50 (the concentration causing 50% inhibition) of 5.3 microM. It also inhibited the calmodulin/Ca2+-stimulated enzymes, with IC50 values of 1.6 and 0.1 microM for myosin light chain kinase (MLCK) and phosphodiesterase respectively. Analysis of inhibition by R-24571 of PL-Ca-PK and MLCK revealed complex kinetics, suggesting that the agent interacted with the cofactors, the enzyme, and/or the cofactor-enzyme complexes. At saturating concentrations of the cofactors, R-24571 inhibited PL-Ca-PK and MLCK noncompetitively with their respective cofactors. Inhibition of MLCK by R-24571 was completely overcome by phosphatidylserine, indicating a strong hydrophobic interaction between R-24571 and the phospholipid in the presence of calmodulin. R-24571 also inhibited phosphorylation of various endogenous proteins in brain stimulated specifically by phosphatidylserine/Ca2+ or calmodulin/Ca2+. The present findings inducated that R-24571 has little specificity in inhibiting two types of Ca2+-dependent protein kinases sensitive to phospholipid or calmodulin.

Animals↗

Stability of phase recognition in complex spatial waveforms.

Observers viewed 200 msec presentations of gratings containing first (0.5 c/deg) and third (1.5 c/deg) harmonic components. The phase of the third harmonic and the absolute position of the grating on the screen varied randomly from trial to trial. Classification of the phase relation (0, 90, 180 or 270 deg was 99% perfect. When a 2 sec period of inspection of the grating or its fundamental preceded the test presentation, strong shifts in perceived waveform were observed that depended on the test grating's position relative to the inspection grating, and resembled the effects seen during continuous viewing. No phase-specific effects were obtained. The pattern of results was exactly that predicted by negative afterimage. Phase recognition at low contrast, and at a high frequency, was also good. Triangular-wave gratings were misperceived (the "square-wave illusion") only when real or simulated afterimages were present. We conclude that recognition of phase relations in a complex waveform is stable when the predictable variation due to afterimages is eliminated.

Afterimage↗

Developmental studies of phospholipid-sensitive Ca2+-dependent protein kinase and its substrates and of phosphoprotein phosphatases in rat brain.

Ontogenetic changes in the protein phosphorylation/dephosphorylation systems in rat brain were investigated. It was found that the activity level of phospholipid-sensitive Ca2+-dependent protein kinase (PL-Ca-PK) in the particulate fraction of grey and white matter and the soluble fraction of grey matter increased rapidly and markedly after birth, reached the highest level at day 30, and declined slightly or remained unchanged thereafter. The enzyme level in the soluble fraction of white matter, in contrast, remained constant throughout the development and maturation of brain. Various ontogenetic changes in the substrate proteins for PL-Ca-PK were also noted. The levels of myelin basic protein and other substrates (notably the Mr 87,000, 58,000, 54,000, and 50,000 protein in grey matter) progressively increased during development, reaching the highest level at adulthood. The level of the Mr 66,000 protein from the particulate fraction of white and grey matter, on the other hand, increased rapidly after birth, reached a peak at day 18, and then declined to the initial neonatal level at the adult stage. The time scale for the increases in the levels of PL-Ca-PK and its many substrates paralleled that of brain development and maturation (synaptogenesis and myelinogenesis). The activity levels of phosphoprotein phosphatases (assayed using 32P-labeled myelin basic protein, histone, and protamine sulfate) were found to only slightly (up to 60%) increase or decrease in certain fractions from different brain regions during development, suggesting that phosphorylation, compared to dephosphorylation, may be more important in determining the phosphorylation state of cellular proteins.

Aging↗

S-100 and other acidic proteins promote Ca2+-independent phosphorylation of protamine catalyzed by a new protein kinase from brain.

A new protein kinase modulated by S-100 (tentatively referred to as protein kinase X) was partially purified from pig brain extracts. The activity of protein kinase X, which was independent of Ca2+, was demonstrated when protamine (free base), but not protamine sulfate and other proteins (including histone), was used as substrate. The enzyme activity, found to distribute in both soluble and particulate fractions and to occur at the highest level in brain compared with other tissues (heart, kidney, liver, skeletal muscle, spleen, and testis) of rats, was also modulated by other acidic proteins (calmodulin, troponin C, and stimulatory modulator) in a Ca2+ -independent manner. S-100 and other acidic proteins appeared to function as "substrate modifiers" by interacting with protamine (a highly basic protein), but not with the enzyme, thus rendering protamine in the complex a superior phosphate acceptor. The two isoforms of S-100 (i.e., a and b) were equally effective. Although the enzyme was not inhibited by many agents (trifluoperazine, melittin, cytotoxin I, polymyxin B, and spermine) shown to inhibit markedly phospholipid/Ca2+- or calmodulin/Ca2+ -stimulated protein kinase, gossypol was found to inhibit specifically protein kinase X. The present findings suggest that S-100, a major acidic protein specific to nervous system, may promote phosphorylation by protein kinase X of certain neural proteins resembling protamine or containing protamine-like domains, in addition to its presumed role of a low-affinity Ca2+ -binding protein.

Animals↗

Phospholipid-sensitive Ca2+ -dependent protein kinase preferentially phosphorylates serine-115 of bovine myelin basic protein.

Phospholipid-sensitive Ca2+ -dependent protein kinase (PL-Ca-PK) and cyclic AMP-dependent protein kinase (A-PK) both preferentially phosphorylated serine residues of bovine myelin basic protein (MBP). Tryptic peptide maps of MBP phosphorylated by PL-Ca-PK or A-PK, however, revealed different phosphopeptides, suggesting a difference in the intramolecular substrate specificity for the two enzymes. Serine-115 of MBP, in the sequence (-Arg-Phe-Ser(115)-Trp-), was found to be a preferred and probably major phosphorylation site for PL-Ca-PK. Because serine-115 of bovine MBP corresponds to serine-113 of rabbit MBP, an in vivo phosphorylation site reported by Martenson et al. (1983), and PL-Ca-PK is present at a very high level in brain and myelin, it is suggested that the enzyme may be responsible for the in vivo phosphorylation of this and other sites in MBP.

Amino Acid Sequence↗

Polyamines inhibit phospholipid-sensitive and calmodulin-sensitive Ca2+-dependent protein kinases.

Effects of polyamines on various protein kinases were investigated. It was found that both phospholipid-sensitive Ca2+-dependent protein kinase and myosin light-chain kinase (a calmodulin-sensitive species of Ca2+-dependent protein kinase) were inhibited to different degrees by polyamines, with an approximate order of inhibitory potency of spermine = 1, 12-diaminododecane greater than spermidine = 1, 10-diaminodecane much greater than cadaverine = putrescine. Kinetic analysis revealed that spermine inhibited the phospholipid-sensitive enzyme non-competitively with respect to Ca2+ (Ki = 0.84 mM) and phosphatidylserine (Ki = 0.90 mM); it also inhibited myosin light-chain kinase non-competitively with respect to Ca2+ (Ki = 1.82 mM) and calmodulin (Ki = 2.73 mM). 1, 12-Diaminododecane, in comparison, inhibited the phospholipid-sensitive enzyme competitively with respect to Ca2+ (Ki = 0.45 mM) and phosphatidylserine (Ki = 0.50 mM); it also inhibited myosin light-chain kinase competitively with respect to calmodulin (Ki = 0.63 mM) but non-competitively with respect to Ca2+ (Ki = 1.49 mM). Moreover, spermine (0.5 mM) was found to inhibit markedly phosphatidylserine/Ca2+- and calmodulin/Ca2+-stimulated phosphorylation of endogenous proteins in rat brain particulate fraction. All the polyamines tested were practically without effect on cyclic AMP-dependent and cyclic GMP-dependent protein kinases. Polyarginine, like spermine, was found to be a more selective inhibitor of Ca2+-dependent protein kinases, whereas polyglutamate preferentially inhibited the cyclic nucleotide-dependent enzymes. The present results indicated that, in addition to certain lipophilic compounds (such as trifluoperazine, palmitoylcarnitine, adriamycin and naphthalenesulphonamide) and polypeptides with hydrophobic regions (such as melittin and polymyxin B) previously reported, polycationic compounds (exemplified by polyamines) could also inhibit the two classes of Ca2+-dependent protein kinases requiring either phospholipid or calmodulin as a cofactor. Because of the high cellular concentration (up to 10 mM) and the differential effects of polyamines, it is suggested that spermine, and to smaller extents spermidine and putrescine, may be involved in the regulation of certain Ca2+-dependent protein-phosphorylation systems in vivo.

Animals↗

Comparative abilities of lanthanide ions La3+ and Tb3+ to substitute for Ca2+ in regulating phospholipid-sensitive Ca2+-dependent protein kinase and myosin light chain kinase.

Although lanthanide ions La3+ and Tb3+ were only slightly able to substitute for Ca2+ to activate phospholipid-sensitive Ca2+-dependent protein kinase (PL-Ca-PK), they potentiated the ability of a suboptimal concentration of Ca2+ to stimulate the enzyme. In comparison, the lanthanides were more effective Ca2+ substitutes for myosin light chain kinase, a calmodulin-sensitive Ca2+-dependent protein kinase. Both enzymes, however, were inhibited by high concentrations of lanthanides either in the presence or absence of Ca2+. Similar effects of the lanthanides were also noted on phosphorylation of endogenous substrates in the particulate fraction of rat brain stimulated by either phosphatidylserine/Ca2+ or calmodulin/Ca2+. The La3+- or Tb3+-stimulated activity of PL-Ca-PK, as the Ca2+-stimulated activity, was inhibited by various agents, such as trifluoperazine, polymyxin B, cobra cytotoxin I, melittin, and spermine.

Animals↗