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Query: UNIPROT:Q86TM3 (cage)
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Intact bovine brain clathrin triskelia, comprising three heavy and three light chains, require either 2 mM calcium or the assistance of protein co-factors for efficient assembly into regular cage structures (Keen, J. H., Willingham, M. C., and Pastan, I. (1979) Cell 16, 303-312). In contrast light chain-free heavy chains assemble readily in the absence of co-factors or calcium. Reconstitution of intact clathrin from heavy and light chains restores the calcium requirement. Our data indicate that light chains impede assembly by creating a kinetic trap rather than by perturbing the affinity of heavy chains for each other. This property suggests a function for light chains as regulatory subunits for clathrin assembly.
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PMID:Bovine brain clathrin light chains impede heavy chain assembly in vitro. 206 36

1. In a 6 x 7 factorial experiment using 2688 22-week-old laying hens of the Lohmann-SL strain kept in cages (4 birds/cage), diets containing six calcium (20, 25, 30, 35, 40, 45 g calcium/kg) and seven phosphorus concentrations (3.2, 4.2, 5.2, 6.2, 7.2, 8.2, 16.2 g total phosphorus/kg (Pt)) were combined orthogonally. The resulting 42 treatments were replicated 8 times so that a replicate consisted of a double cage of 2 x 4 hens. The experiment lasted 40 weeks (10 x 28 days). 2. The experimental diets, based on maize and soyabean meals contained 11.5 MJ metabolisable energy/kg and 175 g/kg protein. Different dietary calcium and phosphorus contents were obtained by substituting oat hulls with limestone and dicalcium phosphate. 3. Mortality, egg production, egg weight, egg mass, food intake and food conversion efficiency were determined as well as the breaking strength, thickness of shells and the percentage of eggs with defective shells. 4. All responses measured were significantly influenced by the variance sources (calcium, phosphorus, interaction). Most of the production traits responded asymptotically to increasing dietary phosphorus concentration, the greatest increases or decreases generally being seen between 3.2 and 5.2 g Pt/kg. Further but weaker increases were seen between 5.2 and 8.2 or 16.2 g Pt/kg. 5. Increases in dietary calcium content always resulted in curvilinear responses. In all cases optimal effects were obtained with diets containing 25 g calcium/kg and the worst values at 45 g calcium/kg. The interaction between calcium and phosphorus was recognised by strong performance depressions and a high mortality at combinations of the lowest phosphorus concentration (3.2 g/kg) with high calcium contents (35 to 45 g/kg). These were largely offset by increasing dietary phosphorus. Thus, between 7.2 and 16.2 g Pt/kg and 25 and 45 g Ca/kg a plateau was formed where only small differences in egg production were observed. 6. From the three egg shell characteristics measured, breaking strength and shell thickness responded differently to the percentage of eggs with defective shells. While breaking strength and shell thickness were respectively negatively and positively influenced by increasing dietary phosphorus and calcium contents, both elements affected the proportion of eggs with defective shells.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Evaluation of the dietary interaction of calcium and phosphorus in the high producing laying hen. 224 45

The biomechanical and biochemical responses of lumbar vertebral bodies during a 12.5-day spaceflight (Cosmos 1887 biosatellite) were determined for rapidly growing rats (90-day-old, Czechoslovakian-Wistar). By use of age-matched vivarium controls (normal cage environment) and synchronous controls (simulated flight conditions), as well as a basal control group (killed before lift-off on the 1st day of flight), the combined influences of growth and space-flight could be examined. Centra of the sixth lumbar vertebrae (L6) were compressed to 50% strain at a fast strain rate while immersed in physiological buffer (37 degrees C). The body masses of vivarium and synchronous controls were significantly heavier than either the flight or basal controls. The flight group had an L6 vertebral body compressional stiffness that was 39% less than the vivarium controls, 47% less than the synchronous control, and 16% less than the basal controls. In addition, the average initial maximum load of the flight L6 was 22% less than vivarium controls and 18% less than the synchronous controls, whereas the linear compressional load of the flight group averaged 34% less than the vivarium and 25% less than the synchronous groups. The structural properties of the vertebrae from the 12.5-day-younger basal group closely resembled the flight vertebrae. Calcium, phosphorous, and hydroxyproline concentrations were not significantly different among the groups. Nevertheless, the lack of strength and stiffness development in spaceflight, coupled with a smaller proportion of mature hydroxypyridinoline cross-links, suggested that the 12.5 days of spaceflight slowed the maturation of trabecular bone in the vertebral bodies of rapidly growing rats.
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PMID:Spaceflight effects on biomechanical and biochemical properties of rat vertebrae. 236 Jun 84

The mechanisms that underlie synaptic plasticity have been largely inferred from electrophysiological studies performed at sites remote from synaptic terminals. Thus the mechanisms involved in plasticity at the secretory sites have remained ill-defined. We have now used somatic synapses of cultured Helisoma neurones to directly assess presynaptic ion conductances and study the secretory apparatus. At these synapses we determined the actions of a modulatory neuropeptide, Phe-Met-Arg-Phe-NH2 (FMRFa), on the release of the neurotransmitter acetylcholine (ACh). Using voltage- and calcium-clamp techniques, we have demonstrated that FMRFa causes a presynaptic inhibition of ACh release by (1) reducing the magnitude of the voltage-dependent calcium current, and (2) regulating the secretory apparatus. The photolabile calcium cage, nitr-5 (refs 3-8), was dialysed into the presynaptic cell. In response to ultraviolet light, calcium was released from nitr-5 and ACh secretion was stimulated. Under conditions of constant internal calcium, FMRFa reduced the rate of ACh release. Thus we conclude that FMRFa reduces the influx of calcium during the action potential and decreases the sensitivity of the secretory apparatus to elevated internal calcium, thereby contributing to a presynaptic inhibition of transmitter release.
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PMID:A neuromodulator of synaptic transmission acts on the secretory apparatus as well as on ion channels. 247 76

Small-cell carcinoma of the lung is a highly lethal form of cancer associated with a wide variety of paraneoplastic syndromes. Using the patch-clamp technique, we have directly demonstrated the presence of voltage-gated K+, Na+, and Ca2+ channels in three cell lines of human small-cell carcinoma, NCI-H128, NCI-H69, and NCI-H146. Whole-cell currents were measured from the tumor cells held at -80 mV and depolarized to -60 to +120 mV. Outward K+ current (IK), which was found in every cell tested, reached 1.58 +/- 0.12 nA (mean +/- SE, n = 24 cells) for H128 cells and 2.14 +/- 0.18 nA (n = 41) for H69 cells in response to a test potential of +80 mV. Unlike H69 and H128 tumor cells, IK from H146 cells occasionally exhibited partial inactivation during the 60-ms pulse length and reached 0.94 +/- 0.15 nA (n = 18) in response to a +80 mV test potential. IK from each of the cell lines was significantly reduced by 4-aminopyridine and tetraethylammonium. The rapidly inactivating inward Na+ current (INa), recorded in H146 cells and about 30% of the H69 and H128 cells tested, demonstrated a peak amplitude of 58 +/- 6 pA (n = 11) at 0 mV and a reversal potential of 47 +/- 2 mV (n = 11). Externally applied tetrodotoxin quickly suppressed INa. For the H128 and H69 tumor cells, inward Ca2+ current (ICa), observed in about 25% of the cells exposed to 10 mM [Ca2+]o, peaked at 5.1 +/- 0.4 ms (n = 5) with an amplitude of 46 +/- 14 pA (n = 5) at +20 mV and partially inactivated over the 40-ms depolarization. In H128 cells exposed to isotonic Ba2+ (110 mM), inward currents with time courses similar to those of ICa were recorded. Nearly all H146 tumor cells demonstrated a significant inward Ca2+ current which peaked with an amplitude of 93 +/- 16 pA (n = 26) at +30 to +40 mV in the presence of 10 mM [Ca2+]o. Application of test potentials 2 s in duration revealed that H146 ICa inactivated in a voltage-dependent manner with a time constant on the order of seconds. Adjustment of the holding potential from -80 mV to -40 mV had no observable effect on the amplitude of the evoked current. These voltage-dependent ion channels may have integral roles in several small-cell carcinoma bioelectric phenomena, including secretion, resting membrane potential, and action potential generation.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Voltage-dependent ion channels in small-cell lung cancer cells. 247 49

EDTA-fluorocarbon microspheres (EDTAFM), calcium disodium ethylene diaminetetraacetate (CaNa2EDTA), calcium- or zinc-diethylene triamine pentaacetate (Ca- or Zn-DTPA) were investigated for their ability to treat experimental lead intoxication in mice. The 48 ICR mice were divided into six groups. Group I = no treatment; The other groups were injected with single ip doses of 210Pb (10 mg Pb2+ +555 kBq/kg). After 24 h they were injected in the tail vein with the chelating agents (20 mg/kg) or an equal volume of 10% glucose (10 mg/kg). Each mouse was housed in one metabolic cage, and urine was collected daily for 3 d. After 3 d, the mice were sacrificed for comparison of lead distribution within the liver, kidney, femur and the entire carcass as measured by 0.047 Mev gamma emission from 210Pb. The results reveal that injection of EDTA-FM to lead poisoned mice pretreated with 210Pb was more effective than Zn- or Ca-DTPA and CaNa2EDTA in reducing the lead induced inhibition in the activity of blood ALAD, and that it increased the excretion of 210Pb into the urine. The hepatic, renal and femur 210Pb contents after treatment with EDTAFM were much more decreased than Zn- or Ca-DTPA and CaNa2-EDTA. The order of effectiveness was EDTAFM greater than Zn-DTPA greater than Ca-DTPA greater than CaNa2-EDTA.
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PMID:[Scavenging effect of EDTA-fluorocarbon microspheres on 210lead]. 251 47

Three intravenous bisphosphonates were compared in the treatment of cancer-associated hypercalcaemia. 48 patients were randomly allocated to one of three treatment groups (each with 16 subjects)--30 mg pamidronate or 600 mg clodronate, both as single intravenous infusions; or etidronate as three infusions of 7.5 mg/kg per day for three consecutive days. Patients were rehydrated with normal saline before bisphosphonate treatment. All three bisphosphonates lowered serum calcium by inhibiting bone resorption; pamidronate was the most potent in this respect. By comparison with the other groups, more patients in the pamidronate group became normocalcaemic, and the effect on serum calcium was apparent sooner and lasted longer.
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PMID:Comparison of three intravenous bisphosphonates in cancer-associated hypercalcaemia. 196 61

Thirty-nine patients with cancer-associated hypercalcaemia were randomly allocated to receive aminohydroxypropylidene diphosphonate (APD), mithramycin, or corticosteroids and salmon calcitonin. Corticosteroids/calcitonin had the fastest calcium-lowering effect, owing mainly to an acute reduction in renal tubular calcium reabsorption; continued therapy over 9 days failed to suppress accelerated bone resorption, however, and most patients remained hypercalcaemic. Mithramycin also substantially reduced serum calcium within 24 h. A further dose on day 2 generally controlled hypercalcaemia until day 6 by reducing both bone resorption and renal tubular calcium reabsorption. By day 9, however, about 50% of the mithramycin-treated patients had started to relapse as bone resorption increased again. With APD serum calcium levels fell more slowly but progressively owing to effective suppression of bone resorption; by day 9 the control of hypercalcaemia was significantly better than in the other treatment groups. Symptoms of hypercalcaemia were greatly relieved, especially by APD.
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PMID:Comparison of aminohydroxypropylidene diphosphonate, mithramycin, and corticosteroids/calcitonin in treatment of cancer-associated hypercalcaemia. 286 17

In the present study protein overlays were used to study the molecular interactions of clathrin with clathrin coat-associated proteins. Coated vesicles (CV) were isolated, subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and transferred to nitrocellulose. The transfers were quenched and equilibrated in buffer, containing 1% Triton X-100. Alpha-Actinin and calmodulin, proteins known to interact with coated vesicles, were iodinated, placed in buffer, and incubated over the transfer for 1 h. After being rinsed extensively, the total amount of 125I associated with the filters was measured, and the filters were then processed for autoradiography. For alpha-actinin, the clathrin heavy chain and a series of lower molecular weight proteins were labeled. The binding of 125I alpha-actinin was inhibited with cold ligand and selectively released from the transfer with a buffer know to strip alpha-actinin from plasma membrane preparations. For 125I calmodulin the predominant binding site was also the clathrin heavy chain. Cold ligand inhibited binding and 60% of the detectable binding were calcium dependent. In addition, when these ligands were used in competition with each other, no significant inhibition was detected in the amount of binding associated with the clathrin heavy chain. These studies show that the clathrin heavy chain is a primary site of the clathrin cage receptive to intracellular interactions and furthermore suggest that the clathrin heavy chain consists of domains of biochemical specificity which may selectively affect the activities of coated vesicles.
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PMID:Evidence for the interaction of alpha-actinin and calmodulin with the clathrin heavy chain. 286 5

A serially transplantable perianal gland carcinoma (CAC-9) was developed in nude mice from a hypercalcemic dog that has been maintained through passage 20. Tumor doubling rate of CAC-9 was 3.1 +/- 0.4 days. Mithramycin (MMC) injected intraperitoneally (8 mg/kg) into nude mice bearing CAC-9 markedly decreased the tumor volume 2 weeks post-injection. MMC returned the elevated serum and urine calcium levels in mice with CAC-9 back to similar values as controls. The few remaining viable tumor cells after MMC were large and had numerous aggregations of intermediate filaments that displaced cytoplasmic organelles. Histomorphometric evaluation of lumbar vertebrae reveled no significant differences in bone resorption of nude mice bearing CAC-9 compared to saline-treated controls. This rapidly growing tumor line in nude mice associated with mild hypercalcemia will be a useful animal model to evaluate combinations of chemotherapy for cancer-associated hypercalcemia.
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PMID:Effects of mithramycin on transplantable canine perianal gland carcinoma (CAC-9) in nude mice: biochemical, histomorphometric, and ultrastructural investigations. 294 19


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