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PubMed1980

Miniaturized whole blood ADCC assay of depressed effector activity in cancer patients.

The ADCC test has been miniaturized to use 0.05 ml of whole blood per test as ADCC effectors. A linear dose response was shown by dilutions of 0.05 ml of blood down to a 1:16 dilution. ADCC effector activities of 40 patients with lung cancer and four with leukemia were markedly less than the activity of 200 normal subjects. Melanoma patients did not have comparable statistically significant loss of ADCC activity.

Antibody-Dependent Cell Cytotoxicity↗
PubMed1986

A study of halothane-diethyl ether azeotrope and the Oxford miniature vaporizer.

The performances of an Oxford miniature vaporizer (OMV) and a Fluotec Mark III vaporizer filled with an azeotropic mixture of halothane and diethyl ether were studied. Gas concentrations were estimated using an EMMA gas analyser and a MIRAN spectrophotometer. Calibration tables for both vaporizers were derived. In a small clinical series with air as the carrier gas, to which a small amount of oxygen was added, the azeotrope was found to be a satisfactory anaesthetic agent, giving a short awakening time and an almost pain-free postoperative course.

Anesthesia, Inhalation↗
PubMed1979

Detection of hepatitis B surface antigen with the miniature centrifugal fast analyzer. A modified reversed passive hemagglutination procedure.

A modified reversed passive hemagglutination test for the detection of hepatitis B surface antigen HBsAg is described. Sera and reagent cells coated with antibody to HBsAg (anti-HBs) are loaded separately into the rotor of a miniature centrifugal fast analyzer. The rotor is centrifuged briefly to transfer the components into its cuvettes. After mixing, the suspensions are allowed to stand at room temperature for 30 min, following which the rotor is again centrifuged and the absorbance of each cuvette is monitored. Cells suspended in serum containing HBsAg leave the light path more rapidly than cells suspended in sera free of antigen. The magnitude of change in absorbance varies directly with the concentration of the antigen. In 45 sera tested by the conventional V-plate technique, findings were as follows: 21 positive, 19 false positive and 5 negative. The automated procedure unequivocally differentiated the 21 positives; results for the false positive and negative specimens were identical and clearly distinguishable from the positive results. The automated procedure enhances specificity, offers equivalent sensitivity, and results that are quantitative and objective.

Absorption↗
PubMed1991

Mast Cell Degranulating Peptide Increases the Frequency of Spontaneous Miniature Postsynaptic Currents in CA3 Rat Hippocampal Neurons.

Mast cell degranulating peptide (MCDP) is a neurotoxic agent isolated from bee venom. It produces a long-term potentiation in the hippocampus. We now report that MCDP, at nanomolar concentrations, induces a reduction of a transient voltage-dependent potassium current (ID) in CA3 rat pyramidal neurons and a persistent (>30 min) enhancement of the frequency of spontaneous miniature excitatory and inhibitory postsynaptic currents (m.e.p.s.c.s. and m.i.p.s.c.s.). M.e.p.s.c.s. and m.i.p.s.c.s. were recorded in the presence of bicuculline (30 microM) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM), respectively. The increased frequency of m.e.p.s.c.s. (408 +/- 60%) and m.i.p.s.c.s. (583 +/- 553%) was independent of the reduction of ID because 4-aminopyridine (4-AP, 30 microM - 2 mM) blocked ID but had no effects on m.e.p.s.c.s. and m.i.p.s.c.s. In the presence of the calcium channel blocker manganese (3 mM), MCDP still enhanced the frequency of m.e.p.s.c.s. (326 +/- 162%). It is concluded that MCDP augments the release of excitatory and inhibitory transmitter by an action, which is independent of calcium influx, through voltage-dependent channels.

Journal Article↗
PubMed1995

Miniature endplate current kinetics at the mouse neuromuscular junction: effects of temperature and medium viscosity.

The time course of miniature endplate currents (MEPCs), derived by extracellular focal recording, was studied in the mouse neuromuscular junction at different temperatures and medium viscosities, and in eserine-treated endplates. At low temperatures (6-10 degrees C), almost the whole MEPC decay is exponential and the rising phase is not significantly modified by the channel closing process. At physiological temperatures (38-40 degrees C), the early part of the decay is much slower than the later part and the rising phase is made shorter and smaller by channel closing, showing that the channel opening and channel closing processes overlap remarkably. Even at physiological temperatures, however, the late part of the MEPC decay shows an exponential time course. At high temperatures the channel opening process has low temperature sensitivity and slows down when bath solution viscosity is increased, suggesting that at high temperatures channel opening kinetics may mainly be controlled by the time course of acetylcholine diffusion. The lower limit of conformational change rate leading to channel opening was estimated at 10 degrees C (4400 s-1). Experimentally recorded MEPCs were mathematically simulated to obtain a quantitative description of the processes controlling MEPC generation. Mathematical simulation further suggests that (i) acetylcholine diffusion kinetics may affect the onset rate of MEPCs without, however, being rate-limiting; and (ii) partial, transient acetylcholinesterase inhibition operated by acetylcholine may explain the low temperature sensitivity exhibited by the onset rate of MEPCs at high temperatures.

Acetylcholine↗
PubMedDate not supplied

Suspected ivermectin toxicosis in a miniature mule foal causing blindness.

A 9-week-old miniature mule foal presented to the Veterinary Medical Teaching Hospital for acute blindness, ataxia, and depression following an overdose of an over-the-counter ivermectin-based de-worming medication. Ophthalmic examination and electrodiagnostic evaluation eliminated outer retinal abnormalities as the primary cause of the bilateral blindness, implicating instead a central neurologic effect of the drug. With symptomatic and supportive care, the foal recovered fully and regained its vision.

Animals↗
PubMed1998

Differential effects of noradrenaline on evoked, spontaneous and miniature IPSCs in rat cerebellar stellate cells.

1. The modulation by noradrenaline (NA) of synapses among stellate cells was investigated in rat cerebellar slices by using presynaptic loose cell-attached recording and postsynaptic whole-cell recording. 2. NA increased the frequency of spontaneous IPSCs recorded from stellate cells without changing their mean amplitude. 3. NA increased the firing rate of stellate cells. This effect persisted after blocking ionotropic glutamate receptors and GABA receptors, indicating that it was independent of synaptic input. 4. The effects of NA on action potential frequency were mimicked by the beta-receptor agonist isoprenaline but not by the alpha-receptor agonist 6-fluoro noradrenaline, and they were not blocked by the alpha-receptor antagonist phentolamine, indicating that they were mediated by beta-receptors. 5. In paired recordings of connected stellate cells, NA slightly decreased the success rate of synaptic transmission. A small decrease in mean IPSC amplitude (excluding failures) and a slight increase in latency were also observed in NA. 6. These results show that, while NA increases the number of action potential-dependent IPSCs by increasing the firing rate of stellate cells, it actually reduces the probability of evoked release. Since previous studies showed that NA increases the rate of miniature IPSCs in this preparation, we conclude that different mechanisms underly the modulation by NA of action potential-dependent and action potential-independent transmitter release.

Action Potentials↗
PubMed2001

Spontaneous miniature outward currents in mechanically dissociated rat Meynert neurons.

1. Spontaneous miniature outward currents (SMOCs) were observed in mechanically dissociated rat Meynert neurons using nystatin perforated patch recordings under voltage-clamp conditions. 2. SMOCs were blocked by apamin, a selective blocker of small conductance Ca(2+)-activated K(+) (SK) channels, but not by blockers for other types of Ca(2+)-activated K(+) channel. 3. Ryanodine (10-100 microM) reduced both the amplitude and frequency of SMOCs. Caffeine (1 mM) increased the SMOC frequency. Blockers of the sarco/endoplasmic reticulum Ca(2+)-ATPase completely abolished SMOCs, indicating a requirement for functioning sarco/endoplasmic reticulum (SR/ER) Ca(2+) stores. 4. Both Cd(2+)-containing and Ca(2+)-free solutions partially inhibited SMOC frequency, a result which suggests that Ca(2+) influx contributes to, but is not essential for, SMOC generation. 5. Thus, SMOCs are SK currents linked to ryanodine- and caffeine-sensitive SR/ER Ca(2+) stores, and are only indirectly influenced by extracellular Ca(2+) influx. The development of this new, minimally invasive mechanical dissociation method has revealed that SMOCs are common in native CNS neurons.

Animals↗
PubMed2001

Oxytocin retrogradely inhibits evoked, but not miniature, EPSCs in the rat supraoptic nucleus: role of N- and P/Q-type calcium channels.

We previously reported that oxytocin (OXT), released from the dendrites of magnocellular neurons in the supraoptic nucleus (SON), acts retrogradely on presynaptic terminals to inhibit glutamatergic transmission. Here we test the hypothesis that oxytocin reduces calcium influx into the presynaptic terminal. We used nystatin perforated-patch recording in vitro to first identify the calcium channels involved in glutamatergic transmission in the SON. [omega]-Conotoxin GVIA ([omega]-CTx) and [omega]-Agatoxin TK ([omega]-Aga) both reduced evoked EPSC amplitude, while nicardipine and nickel had no effect. A combination of [omega]-CTx and [omega]-Aga completely abolished the evoked EPSCs. This depressant effect was accompanied by an increase in the paired pulse ratio with no change in the kinetics of the evoked EPSCs, AMPA currents or postsynaptic cell properties. These results suggest that presynaptic N- and P/Q-type calcium channels mediate glutamate release in the SON while L-, T- and R-type channels make little or no contribution. Oxytocin-induced reduction of the evoked EPSC was substantially occluded in the presence of [omega]-CTx but only partially in the presence of [omega]-Aga. Amastatin, an endopeptidase inhibitor that increases the level of endogenous OXT, also reduced the evoked EPSC. This amastatin effect was also occluded by [omega]-CTx and [omega]-Aga. Miniature EPSCs, which are independent of extracellular calcium, were unaffected by either [omega]-CTx or by OXT, thus further substantiating an action of both compounds on calcium channels. Therefore, dendritically released oxytocin acts mainly via a mechanism involving the N-type channel, and to a lesser extent the P/Q-type channel, to decrease excitatory transmission.

Agatoxins↗
PubMed1981

Chloroquine intoxication induces ganglioside storage in nervous tissue: a chemical and histopathological study of brain, spinal cord, dorsal root ganglia, and retinal in the miniature pig.

The effect of chronic chloroquine intoxication on lipid composition, particularly the gangliosides, was studied in the nervous system of miniature pigs, type Göttingen. The tissues examined were cerebrum, spinal cord, dorsal root ganglia and retina. Chloroquine was given in the diet in doses of 2.0-3.5 g/kg good. The intoxication of the pigs was started at the age of 100-240 days and continued for 177-219 days. The control pigs received the same diet without chloroquine. The ganglioside concentration was increased in all the tissues examined. Dorsal root ganglia and retinal were the tissues affected most and showed a twofold increase. This corresponded to the light and electron microscopically demonstrated extensive storage process in the perikarya of dorsal root ganglion cells and inner ganglion cells of the retinal. Under light microscopy the storage material was granular, intensely PAS-positive and dissolved by paraffin embedding. The electron microscopical equivalent consisted of conglomerates of membranous lysosomal residual bodies. In cerebrum the ganglioside concentration was increased by 12%. Storage in the brain varied widely between different systems and types of cells. The allocortex was much more affected than the isocortex. Certain inhibitory ganglion cell types, such as the basket cells, exhibited the most massive storage of all. The spinal medulla was morphologically less involved but showed approximately the same ganglioside increase, though not statistically significant. With the exception of cerebrum the increase in the tissues examined involved all the individual gangliosides, most severely ganglioside GM2 and three fucogangliosides. In cerebrum only the ganglioside GM2 was increased more than the other gangliosides. Chloroquine intoxication did not affect the concentration of phospholipids or cholesterol in the cerebrum, spinal cord or dorsal root ganglia, but in retina the acidic phospholipids were significantly increased.

Animals↗
PubMed1984

Effects of sea-anemone toxin (ATX-II) on the frequency of miniature endplate potentials at rat neuromuscular junctions.

Soleus and extensor digitorum longus muscles were isolated from rats. The muscles were exposed to ATX-II, a toxin isolated from extracts of the sea-anemone Anemonia sulcata . The toxin caused a dose-dependent increase in the frequency of miniature endplate potentials in both types of muscle. The increase in frequency could be reversed by the application of tetrodotoxin (TTX), and could be prevented by prior exposure of the muscles to TTX. It is concluded that ATX-II causes a sodium-dependent depolarization of the nerve-terminal membrane.

Action Potentials↗
PubMed1988

Early induction by crotoxin of biphasic frequency changes and giant miniature endplate potentials in frog muscle.

1. Following the addition of crotoxin (250 nM) at the frog neuromuscular junction, there was an initial fall in frequency of miniature endplate potentials (m.e.p.ps), followed by a secondary rise which was characterized by the appearance of large spontaneous potentials (giants, g.m.e.p.ps) and an occasional large potential of the burst type. 2. In the presence of 2-(4-phenylpiperidino)cyclohexanol (AH5183, vesicamol), an inhibitor of vesicular acetylcholine uptake, the frequency of g.m.e.p.ps induced by crotoxin was reduced. 3. The characteristic changes in m.e.p.p. frequency and amplitude distribution were absent with crotoxin in Sr-EGTA Ringer. In the presence of high concentrations of Mn (3.6 or 5.4 mM with 0.9 mM Ca), the crotoxin-induced initial fall and the onset of the secondary rise in m.e.p.p. and g.m.e.p.p. frequencies were slower. The timing of these phases was unaffected by Ca concentrations ranging from 6.3 to 0.9 mM. 4. High concentrations of Mn ions partially inhibited the phospholipase A2 activity of crotoxin on artificial phospholipid membranes. This also supports the involvement of the Ca-dependent phospholipase A2 subunit in both phases of the physiological action of the toxin. 5. G.m.e.p.ps were associated with a moderate increase in m.e.p.p. frequency (2-3 s-1) and were of a time-course similar to that of m.e.p.ps. They persisted after washing with medium lacking Ca ions and in the presence of Ca-Mn Ringer that blocked evoked responses. 6. It is concluded that crotoxin, acting through its phospholipase A2 subunit, produces specific disturbances of synaptic exocytosis and vesicle formation in the axolemma of the motor nerve terminal which lead to biphasic changes in m.e.p.p. frequency and the onset of large spontaneous potentials.

Animals↗
PubMed1991

Depression of miniature endplate potential frequency by acetylcholine and its analogues in frog.

1. Acetylcholine (ACh), 7.5 x 10(-5) M, and carbachol, 5 x 10(-6) M (CCh) depressed the frequency of miniature endplate potentials (m.e.p.ps) in the frog (Rana temporaria) sartorius neuromuscular junction with active acetylcholinesterase to about 50-55% of the controls. 2. A similar depression was produced by the nicotinic agonists, nicotine, suberyldicholine and tetramethylammonium. 3. The muscarinic agonists, oxotremorine, methylfurmethide and methacholine were without effect on m.e.p.p. frequency. The muscarinic antagonist, atropine and the nicotinic antagonist, (+)-tubocurarine, had no effect on the depression of m.e.p.p. frequency evoked by CCh. 4. The ganglionic blockers, benzhexonium and IEM-1119, were also without effect on the CCh-evoked depression of m.e.p.p. frequency. 5. Pretreatment of muscles with anticholinesterases did not prevent the CCh-induced drop in m.e.p.p. frequency. 6. The effect of CCh was proportionally the same as in the controls in preparations where the m.e.p.p. frequency was changed by elevation of K+ and in the presence of theophylline, noradrenaline, dibutyryl adenosine 3':5'-cyclic monophosphate (db cyclic AMP) and db cyclic GMP. 7. An inhibitor of Na+,K(+)-ATPase, ouabain, 5 x 10(-5) mol l-1, prevented or reversed the depression of m.e.p.p. frequency by CCh. However, the depression was present in a nominally K(+)-free medium. Insulin and adrenaline, which are considered to be Na+,K(+)-ATPase activators, were without effect on depression of m.e.p.p. frequency. 8. The depression of m.e.p.p. frequency by 5 x 10(-6) M CCh was the same at temperatures between 5 and 30 degrees C with a Q10 near to 1.0. When threshold amounts of CCh were used (6 x 10-7 and 3 x 10-7 M), the depression was less at higher temperatures.9. The receptive structures responsible for the CCh (or ACh)-evoked depression of m.e.p.p. frequency differ pharmacologically from muscarinic, nicotinic ganglionic and neuromuscular junction ACh-receptors as well as from the synaptic cholinesterase, in contrast to previous reports (Duncan & Publicover, 1979).The low temperature-dependence points to the possibility that physical rather than biochemical processes are limiting in this presynaptic effect of cholinomimetics.

Acetylcholine↗
PubMed1993

Potentiation of miniature endplate potential frequency by ATP in Xenopus tadpoles.

1. Extracellular application of ATP (1 mM), a substance co-stored and co-released with acetylcholine in peripheral nervous systems, potentiated the spontaneous secretion of acetylcholine (ACh) but had no effect on the amplitude and decay time constant of miniature endplate potentials (m.e.p.ps) at neuromuscular synapses in Xenopus tadpoles. 2. alpha,beta-Methylene ATP (0.3 mM) and GTP (1 mM) were also effective in increasing m.e.p.p. frequency. On the other hand, ADP, AMP and adenosine (all at 1 mM) decreased m.e.p.p. frequency. 3. Unlike the transient effect of ATP analogue and GTP on m.e.p.p. frequency, the phorbol ester TPA (2 microM) which is a protein kinase C activator, increased m.e.p.p. frequency consistently and the effects lasted as long as the presence of TPA. 4. Staurosporine (0.5 microM) and H-7 (10 microM), which are protein kinase C inhibitors, each decreased the basal level of m.e.p.p. frequency and markedly inhibited the effects of both ATP and TPA. 5. These results suggest that there is a basal activity of cytosolic protein kinases in the nerve terminals of Xenopus tadpoles and the effect of ATP is probably mediated by the binding of membrane surface purinoceptors which in turn activates cytosolic protein kinases and increases ACh release.

Acetylcholine↗
PubMed2004

Concept for a new hydrodynamic blood bearing for miniature blood pumps.

The most crucial element of a long-term implantable rotary blood pump is the rotor bearing. Because of heat generation and power loss resulting from friction, seals within the devices have to be avoided. Actively controlled magnetic bearings, although maintenance-free, increase the degree of complexity. Hydrodynamic bearings for magnetically coupled rotors may offer an alternative solution to this problem. Additionally, for miniature pumps, the load capacity of hydrodynamic bearings scales slower than that of, for example, magnetic bearings because of the cube-square-law. A special kind of hydrodynamic bearing is a spiral groove bearing (SGB), which features an excellent load capacity. Mock-loop tests showed that SGBs do not influence the hydraulic performance of the tested pumps. Although, as of now, the power consumption of the SBG is higher than for a mechanical pivot bearing, it is absolutely contact-free and has an unlimited lifetime. The liftoff of the rotor occurs already at 10% of design speed. Further tests and flow visualization studies on scaled-up models must demonstrate its overall blood compatibility.

Assisted Circulation↗
PubMed2006

Developmental differences in the sensitivity of spontaneous and miniature IPSCs to ethanol.

BACKGROUND: Ethanol (EtOH) consumption by juveniles and adolescents is an important public health problem. Recent studies have indicated that adolescent animals are less sedated by EtOH than adult animals and experience less motor impairment. Thus, human adolescents may be able to consume more EtOH prior to sedation, putting them at greater risk for EtOH addiction and other negative consequences of EtOH use. However, the mechanisms underlying this developmental difference are unknown. One contributing factor may be gamma-aminobutyric acid-A (GABA(A)) receptor-mediated inhibition, which is known to produce sedation. We have shown that evoked, GABA(A) receptor-mediated inhibitory postsynaptic currents (IPSCs) are less powerfully enhanced by EtOH in neurons from juvenile or adolescent animals than in those from adult animals; however, the mechanisms of this developmental difference in sensitivity are unknown. METHODS: Using whole-cell recording, we tested the response of spontaneous and miniature GABA(A) receptor-mediated IPSCs (sIPSCs and mIPSCs) to EtOH in rat hippocampal slices from animals representing two distinct developmental stages: adolescent and adult. RESULTS: We found significantly greater EtOH-induced enhancement of the frequency of sIPSCs in cells from adult animals compared to those from adolescent animals. Although EtOH also increased the frequency of mIPSCs, this effect was not age dependent. EtOH did not significantly affect the kinetics of mIPSCs. CONCLUSIONS: We conclude that the sensitivity of GABA(A) receptor-mediated inhibitory processes to EtOH increases with development from the adolescent period to adulthood, and that this is likely mediated by developmental changes in the effect of EtOH on interneuron excitation.

Age Factors↗
PubMedDate not supplied

Lateral patellar luxation in miniature horses.

Bilateral luxation of the patella in four Miniature Horses was corrected by a lateral release incision and medial imbrication of the parapatellar fascia to the tendon of the sartorius muscle. Before surgery, the four horses had a grade 3 to 4 lateral patellar luxation bilaterally and had difficulty walking. Trochlear ridge hypoplasia was evident on radiographs in each horse. Follow-up information varied from 11 months to 4 years after surgery. Three horses had no patellar luxation or lameness. The other horse had a normal right stifle, but patellar luxation (grade 3) had recurred on the left.

Animals↗