Urinary inhibitor of the formation of calcium oxalate. Normal urine was examined for substances which inhibited the formation of calcium oxalate, using a separation scheme which involved ultrafiltration, alcohol precipitation, electrophoresis and paper chromatography. The major anionic inhibitors were found to be citric acid, isocitric acid and pyrophosphate. The inhibitory activity of urine and that of an artificial urine having the same concentration of urea, creatinine, sodium, potassiu, magnesium, calcium, ammonia, sulphate, chloride, inorganic phosphate, oxalate, citrate, isocitrate and pyrophosphate, and the same pH were identical. This confirmed that urine does not contain an unidentified potent substance which inhibits the formation of calcium oxalate.


Growth and effect of chlamydiae in human and bovine oviduct organ cultures. Organ cultures of 10 Fallopian tubes were inoculated with a genital strain of Chlamydia trachomatis and seven were infected. Infection was enhanced by centrifuging the organisms on to the tissues, larger numbers of organisms being reisolated from the tissues after this procedure. There was evidence of chlamydial multiplication because the number of organisms which were recovered from the tissues three to five days after inoculation had increased. Recovery was rare, however, after the sixth day, thus suggesting a self-limiting infection. Organ cultures of two bovine oviducts were infected with the bovine abortion strain of Chlamydia psittaci, but in these experiments centrifugation of the inocula did not enhance infection. The organisms were found in both the tissue and medium of cultures up to 18 days after inoculation and in much greater numbers than in the C. trachomatis-infected Fallopian cultures. Chlamydial infection was not entirely host-tissue specific, because C. trachomatis organisms were isolated from bovine oviduct cultures. Inclusions, however, were not detected histologically or electron microscopically in the epithelium of C. trachomatis-infected cultures, but they were detected by these means in C. psittaci-infected bovine cultures. All the elements of the chlamydial growth cycle were seen by electron microscopy, organisms being found in ciliated and possibly non-ciliated cells, and shedding of some infected epithelial cells was observed. No evidence of extensive epithelial cell damage was observed, however, and no loss of ciliary activity was detected in cultures infected with either C. trachomatis or C. psittaci when compared with uninoculated cultures. Thus acute salpingitis, when caused by chlamydial infection, is probably immunologically mediated.


Distribution of PNS myelin proteins and membrane enzymes in fractions isolated by continuous gradient zonal centrifugation. Myelin was purified from adult rabbit sciatic nerve by two procedures: discontinuous gradient centrifugation and continuous gradient zonal centrifugation. Two fractions were obtained from the discontinuous gradient. The fraction floating on 0.32 M sucrose and the fraction recovered from the 0.32/0.85 M sucrose interface showed typical myelin membranes by electron microscopy and typical myelin proteins by gel electrophoresis. The specific activity of 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP) decreased from the top to the bottom of the discontinuous gradient. The myelin separated by zonal centrifugation on a continuous sucrose gradient showed three distinct peaks (on monitoring optical density) at 0.10, 0.30 and 0.57 M sucrose. The latter peak yielded 92% of the material applied. The two minor peaks of low density exhibited high CNP and acetylcholinesterase (AChE) activities but the specific activity of both enzymes increased markedly at the heavy end of the gradient. The zonal fractions showed typical myelin proteins in all fractions by polyacrylamide gel electrophoresis but with important quantitative differences. These results indicate that PNS myelin shows significant heterogeneity.


Evidence that an excitatory connection between the periaqueductal gray and nucleus raphe magnus mediates stimulation produced analgesia. Both electrical stimulation and injection of morphine into the midbrain periaqueductal gray (PAG) produce analgesia in the rat. There is evidence that this analgesic effect is mediated by a descending system that involves nucleus raphe magnus (NRM) and adjacent reticular formation. In the studies reported here, the activity of the cells in the PAG was increased by microinjection of glutamate in this area and its effect on both the activity of single cells in the NRM and on a flexion reflex elicited by noxious heat was measured. It is shown that an increase in the firing rate of the cells in the PAG is associated with a raised threshold for flexion and is also correlated with an increase in the firing rate of a majority of the cells in the NRM. This effect on the flexion reflex can be abolished by (a) lesion of the nucleus raphe magnus and a small area of the reticular formation surrounding this nucleus and (b) by nalazone 20 min after its i.v. injection. It is concluded that there is an excitatory connection between the periaqueductal gray and the nucleus raphe magnus and that activation of this system can cause analgesia.


Role of the nucleus raphe magnus in opiate analgesia as studied by the microinjection technique in the rat. The analgesic effects of morphine (5 microgram, 0.2 microliter) microinjected into the nucleus raphé magnus (NRM) and the surrounding reticular formation of the rat were tested using vocalization after electric shock to the tail as the test for analgesia. Only sites in the NRM produced powerful analgesic effects, strongest analgesia being equivalent to 3 mg/kg i.v. morphine. The analgesia produced by the microinjection was reversed by systemic naloxone. Pretreatment with systemic cinanserin, a blocker of serotonergic receptors, led to a pronounced diminution of the analgesic effects of the morphine. The effects of microinjections of naloxone (5 microgram 0.2 microliter) were studied for their effect on analgesia produced by systemic morphine. The analgesia following 3 mg/kg i.v. morphine was diminished by the microinjection of naloxone but the naloxone almost completely reversed the analgesic effects of 1.5 mg/kg i.v. morphine. These results further substantiate the role of the NRM in analgesic mechanisms.


Endogenous phosphorylation in vitro: differential effects of brain state (anesthesia, post-mortem) on electrophoretically separated brain proteins. In vitro phosphorylation of rat cerebral cortex synaptosomes was measured in animals that had been acutely treated with sodium pentobarbital. [32P]Labelled phosphoproteins were separated by SDS-slab gel electrophoresis, and the autoradiographs were analyzed by densitometry. We report here that Band F of our previous reports can be separated into two components, F1 and F2, using an improved gel system. This separation is particularly relevant in this report since these components appear to be differentially sensitive to the manipulations used. Specifically, we found that while F1 phosphorylation was markedly diminished by deep barbiturate anesthesia, F2 was relatively stable. While phosphorylation of F2 was also stable 24 h post-mortem, Band F1 phosphorylation was no longer detectable. Finally, while osmotic shock treatment of synaptosomes reduced phosphorylation of F2 somewhat, it eliminated the in vitro phosphorylation of Band F1. We found that under light barbiturate anesthesia, just at the time when the animals lost the righting reflex, the in vitro phosphorylation of Bands D (MR 78,000--80,000 daltons), F1 (MR 47,000--49,000) and F2 (MR 40,000--45,000) increased relative to unanesthetized controls. The in vitro labelling of Bands D and F1 was depressed in tissue prepared from animals that were deeply comatose. These effects of pentobarbital were more pronounced when animals were sacrificed by liquid nitrogen immersion, rather than by decapitation. Cyclic AMP-dependent phosphorylation of Band D exhibited remarkable stability 24 h post-morten (7 days in one case), even when brain tissue was left at room temperature (21--23 degrees C). Phosphorylation of Band F1, however, was not detectable in post-mortem tissue. The results of these studies indicate that phosphorylation of Band F1 is: (1) sensitive to pentobarbital, and (2) unstable post-mortem. Previous findings from our laboratory suggest that Band F1 is: (3) increased in phosphorylation in liquid nitrogen P2 preparations, and may be (4) cAMP-independent, (5) rapidly turning over its phosphate in vivo, and (6) altered by a training experience. Other evidence suggests that: (7) Band F1 phosphorylation may be Ca2+-dependent and that: (8) its phosphorylation is sensitive to osmotic lysis of synaptosomes. The results suggest an important and perhaps unique role for Band F1 in neuronal function.


Effects of opiate-like peptides, morphine, and naloxone in the photosensitive baboon, Papio papio. The effects of intracerebroventricular (i.c.v.) or systemic injections of Met- or Leu-enkephalin, beta-endorphin, FK 33.824 (D-Ala2, MePhe4, Met(O5)-ol-enkephalin) and of morphine and naloxone have been studied in baboons, Papio papio, which spontaneously show photically induced epileptic responses. Animals were chronically implanted with epidural or deep recording electrodes and a cannula in one lateral ventricle, and tested whilst seated in a primate chair. In some animals the natural syndrome was enhanced by the prior administration of DL-allylglycine, 100--200 mg/kg, i.v. Met- or Leu-enkephalin, 1--10 mg, i.c.v., did not lead to any manifest focal or generalized seizure discharges. Nor did it lead to any consistent enhancement or reduction of photically induced myoclonic responses (as tested 5--10 min after injection). beta-Endorphin, 0.1--0.5 mg, i.c.v., did not enhance or impair photically induced myoclonic responses. FK 33.824, 0.1--0.5 mg, i.c.v., depressed respiration and slowed EEG background rhythms for 9--15 h. This was associated with a loss of myoclonic responses to photic stimulation. These effects were reversed for 20--40 min following the injection of naloxone, 1 mg/kg i.m. A depression of respiration and a slowing of EEG rhythms was seen beginning 5--20 min after FK 33.824, 2 or 4 mg/kg, i.v. The higher dose also abolished photically induced myoclonic responses. Naloxone, 1 mg/kg, definitively reversed these effects. Morphine, 5--10 mg i.c.v., tended to increase the latency to onset of generalized myoclonus during photic stimulation. Myoclonic responses were delayed or diminished after morphine, 5 mg/kg, i.m. Naloxone, 1--2 mg/kg i.m., reversed this effect. Naloxone, 0.2--5.0 mg/kg i.m., alone, did not significantly modify photically induced myoclonus, either in animals of low or high initial responsiveness, or in those pretreated with allylglycine.


Blockade of progesterone-induced increase in hypothalamic luteinizing hormone-releasing hormone levels and serum gonadotropins by intrahypothalamic implantation of 6-hydroxydopamine. The present study was undertaken to identify the hypothalamic locus where norepinephrine (NE) nerve terminals communicate with luteinizing hormone-releasing hormone (LH-RH)-containing neurons involved in the stimulatory feedback action of gonadal steroids on LH and FSH release. Ovariectomized rats received estradiol benzoate (10 microgram/rat s.c.) on day 0. Intracranial implants containing either 6-hydroxydopamine (6-OHDA), to destroy NE terminals, or cocoa butter (controls) were placed bilaterally in the suprachiasmatic nucleus (SCN), medial basal hypothalamus (MBH) or olfactory tubercle (OT) on day 1. Progesterone (P, 5 mg/rat s.c.) was administered at 10.00 h on day 2 to elicit increases in serum LH and FSH and the MBH LH-RH levels in the afternoon. Implantation of 6-OHDA in the SCN resulted in a marked depletion of NE in and around the region of the SCN in the preoptic-anterior hypothalamic area (POA-AH) without adversely affecting dopamine (DA) concentrations, and blocked the P-induced afternoon increase in the MBH LH-RH and serum gonadotropin levels. Similar reduction in the MBH NE concentrations occurred following placement of 6-OHDA in the MBH; however, these as well as implants in the OT were ineffective in suppressing the P-induced effects. These studies show that functional integrity of the SCN regions is critical in manifestation of the P-induced rise in the MBH LH-RH activity, and this region in the POA-AH, therefore, may be the primary locus of synaptic communication between NE terminals and LH-RH neurons.


Changes in the visual system of monocularly sutured or enucleated cats demonstrable with cytochrome oxidase histochemistry. Endogenous cytochrome oxidase activity within the mitochondria of neurons and neuropil was demonstrated histochemically under normal and experimental conditions. Since enzymatic changes were noted with chronic neuronal inactivity in the auditory system (Wong-Riley et al), the present study sought to examine functionally induced enzymatic changes in the visual system of kittens. Eight kittens were used experimentally: 5 had monocular lid suture for varying periods of time; one had binocular lid suture followed by monocular suture followed by binocular opening; two had monocular enucleation. All initial procedures were performed before eye opening. Materials from other normal kittens and cats were also used as controls. At the end of the experiments, the animals were perfused with aldehyde solutions and frozen sections of the brains were incubated for cytochrome oxidase activity (a detailed protocol was outlined). The results indicated that the deprivation caused by monocular suture produced a decrease in the cytochrome oxidase staining of the binocular segment of the deprived geniculate laminae. Enucleation yielded a greater decrease in the cytochrome oxidase activity in the affected geniculate laminae. However, the staining in the 'normal' lamina extended across the interlaminar border to include a row of surviving large cells in the 'denervated' lamina. The staining of the monocular segment appeared not to be affected by lid suture, but was decreased by enucleation. At the cortical level, lamina IV in area 17 of normal cats was stained darkly as a continuous band. Following lid suture, this pattern was replaced in part by alternating columns of light and dark staining, suggestive of ocular dominance columns. Thus, a decrease in neuronal activity due to reduced visual stimulation or destruction of the primary afferent nerves led to a significant decrease in the level of oxidative enzyme activity one to several synapses away.


Potentiation of the effects of GABA by pentobarbitone. Pentobarbitone (50 microM) has been shown to potentiate GABA-induced responses in rat isolated superior cervical ganglia and in dorsal or ventral root fibres of immature rat isolated spinal cords. Measurements of dose ratios for the antagonism of GABA-induced responses of dorsal root fibres by bicuculline showed that pentobarbitone (50 microM) did not significantly alter this antagonism. This result indicates that the antagonism of GABA by bucuculline at dorsal root fibres is not reversed by pentobarbitone. Ventral root responses to GABA (but not glycine) were also potentiated by pentobarbitone (+/-)-Nipecotic acid (300 microM) potentiated responses to GABA much more than those to 3-aminopropane-sulphonic acid.


Turnover of phosphatidyl choline and changes in enzymatic activity in cell membranes of the CNS during early myelination. Subfractions of fraction A, which floats on top on 0.8 M sucrose in the classical density gradient used for the isolation of brain subcellular fractions (A1 corresponding in adult rats to myelin and A2 which corresponds to the myelin-like fraction), were studied in comparison to other brain subcellular fractions in 5 day old rats and at different stages of development, up to 60 days of age. Variations in the enzymatic activity of acetylcholinesterase, non-specific cholinesterase and 2',3'-Cyclic nucleotide phosphohydrolase, changes in lipid and protein composition and turnover of phosphatidyl choline were investigated. Results indicate that fractions A1 and A2 obtained prior to the beginning of myelination could be composed of fragments of the oligodendroglial cell plasma membranes, and that both fractions undergo substantial changes in chemical composition, enzymatic activity and in turnover of phosphatidyl choline during maturation. In vivo experiments at short times, using radioactive choline as a precursor of phosphatidylcholine suggest that membrane fragments isolated in fraction A2 are precursors of those isolated in fraction A1 at all ages.


The bulbo-spinal indoleaminergic pathway in the frog. The role of the bulbo-spinal indoleaminergic pathway of the frog was investigated. Using the isolated spinal cord preparation, responses of motoneurons of segments 9 and 10 to lateral column stimulation were recorded from ventral roots (LC-VRP) and by intracellularly placed microelectrodes. Control responses were compared to those obtained after addition of substances known to alter indoleaminergic synthesis or receptor activation. Responses from spinal cords of animals that were pretreated with indoleamine-depleting agents were compared to mean control responses. Procedures that inhibited indoleamine synthesis or blocked indoleamine receptors reduced motoneuronal activity. This was manifested as an increase in mean latency and a decrease in amplitude of the monosynaptic LC-VRP, an increased duration of suppression following an LC conditioning stimulus, and a decrease in spontaneous activity. In contrast to intracellularly recorded responses from control cords, those recorded under these conditions typically showed single spikes with longer latencies. Additions of indoleamine precursors to normal cords or to cords depleted of monoamines by reserpine shortened mean latencies, increased amplitudes of LC-elicited responses and caused an increase in spontaneous activity. These observations were recorded both extracellularly and intracellularly. Our results suggest that the bulbo-spinal indoleaminergic pathway modulates the output of motoneurons of the frog spinal cord.


Isotherm of alcohol-cooled cryoprobe.

Isotherm of alcohol-cooled cryoprobe. The spread of cooling from the tip of a cryoprobe placed in the ventral posterior lateral nucleus of the thalamus in 10 cats was determined. A 20 degrees C isotherm was described as having a diameter of 4 mm and extending 2 mm below the tip of the cryoprobe and tapering up along the shaft of the cryoprobe for a distance of 2 mm, when the cryoprobe tip temperature was maintained at 3 degrees C.


Feedback loop between locus coeruleus and spinal trigeminal nucleus neurons responding to tooth pulp stimulation in the rat. Studies were performed to elucidate reciprocal relationships between locus coeruleus (LC) and spinal trigeminal nucleus (STN) neurons responding to tooth pulp (TP) stimulation using rats anesthetized with alpha-chloralose. LC conditioning stimulation inhibited STN field potential as well as orthodromic spike generation of STN neurons produced by ipsilateral TP stimulation, confirming the previous findings in cats that LC neurons played an inhibitory role in the orthodromic transmission in STN neurons. Forty-one out of 56 LC neurons were activated by ipsilateral TP stimulation and 12 neurons by stimulation of both ipsi- and contralateral TP. STN stimulation usually excited LC neurons with a significantly shorter latency than did TP stimulation, including three LC neurons with a latency of less than 2.0 msec. These results indicate the existence of input from TP to LC neurons via multisynapses. In addition, neurons antidromically activated by STN stimulation were found in LC. It is highly probable, therefore, that there is a feedback loop between LC and STN, which might control input from TP to STN.


[Effect of stimulation of alpha and beta presynaptic adrenoceptors on the liberation of noradrenaline produced by electric stimulation in rabbit carotid sinus]. The experiments carried out on the isolated carotid sinus in the rabbit demonstrated that alpha and beta adrenoceptors are capable of detecting the circulating catecholamines and the catecholamines of the synaptic cleft in order to modulate the release of noradrenaline from the sympathetic nerve endings in response to a potential action.


[Cellular receptors of gonadotropic hormones. Immunocytochemical evidence in rat testicle]. We have performed topical applications of HCG and LH on fixed paraffin-embedded sections of adult rat testes. Thereafter, immuno-cytochemical demonstration of the peptides was carried on and demonstrated that HCG and LH specifically bound to the cell membrane of Leydig cells. This technique is highly reproductible provided that adequate fixation (with formalin containing fixatives) as been performed.


[Fate of Leydig cells in testicular organ culture]. In organ culture experiments, the Leydig cells of mouse testes can still be recognized after 12 days at 31 degrees C. HCG binding sites are effectively demonstrated. Meanwhile, when HCG is added at the initiation of the cultures, it is selectively stored in the cytoplasm of the Leydig cells. These findings point to a fairly good survival of Leydig cells in our tissue culture system.


Differences in norepinephrine and dopamine neurotransmitter storage systems. Low doses of d-amphetamine (d-AMP) produced a 50% or greater decrease in the firing rates of both dopamine (DA) neurons (substantia nigra zone compacta) and norepinephrine (NE) neurons (locus coeruleus). However, pretreatment with the tyrosine hydroxylase inhibitor alpha-methyl-para-tyrosine (alpha-MT) blocked the d-AMP-induced reduction in DA neuron firing rate, but had no effect on the d-AMP-induced reduction in NE cell firing rate. Similarly, alpha-MT administered subsequent to d-AMP readily reversed the d-AMP-induced decrease in the firing rates of DA cells, but caused no significant reversal in NE cell firing rates. These electrophysiological findings, in conjunction with biochemical and behavioral data, support the hypothesis that there is a difference in the DA and NE neurotransmitter storage mechanism. In the DA neuron, there appears to be a slow transfer between stored and readily-releasable (newly synthesized) amine pools so that, following synthesis inhibition, there is little DA available for release. However, in the NE neuron, there is a more rapid mobilization of stored amine to readily releasable sites, such that d-AMP continues to cause the release of NE even though synthesis of transmitter is blocked.


Monosynaptic facilitatory pathway from the hypothalamic ventromedial nucleus to the frontal cortex in the rat. Stimulation of the ventromedial nucleus (VMH) evoked a short latency negative wave with two peaks (cN1 and cN2) followed by a small positive wave (cP) at the ipsilateral dorsal frontal cortex (area 10) in the rat. The maximum response was observed from the lateral edge of the frontal pole. From the depth profiles of recordings, cN1 changed polarity at a depth of about 4 mm and the cN1-cN2 changed into a large compound action potential at the medioventral part of the frontal pole at a depth of about 6 mm. Since the surface evoked potential and the compound action potential followed high frequency stimulation, these respective potentials are concluded to be due to antidromic and monosynaptic activation of the cortical neurons. This was verified by unit recording experiments. The cP was concluded to be produced by the initial rise of the monosynaptic EPSP.


Anticonvulsant drug-induced osteomalacia: alterations in mineral metabolism and response to vitamin D3 administration. Parameters of mineral metabolism were examined in 6 patients with moderately severe anticonvulsant drug-induced osteomalacia. Compared to 15 matched controls, the patients exhibited significantly reduced serum calcium, inorganic phosphate, and 25-hydroxyvitamin D concentration, elevated serum alkaline phosphatase and immunoreactive parathyroid hormone (iPTH) concentration, reduced intestinal 47Ca absorption, reduced urinary calcium and increased urinary hydroxyproline excretion, and reduced forearm bone mass. Intestinal absorption of vitamin D3 was normal. Following 4 months of treatment with vitamin D3 (4000 units/day), serum 25-OHD concentration was increased to 3 times mean normal values and all parameters except serum iPTH, urinary calcium excretion, and forearm bone mass were returned to levels not significantly different from normal. Serum iPTH concentration was reduced by 39% (P less than 0.05); 24-h urinary calcium excretion rose by 98% (P less than 0.001), and forearm bone mass increased by 5.6% (P less than 0.05). It is concluded that moderate-dose vitamin D3 supplementation is effective in normalizing parameters of mineral metabolism in this disorder, despite evidence of resistance to the biologic effects of vitamin D.


The effect of morphine on human neuromuscular transmission. By utilizing high frequency nerve stimulation, we observed the effects of morphine sulphate, 0.5 mg.kg-1 on human neuromuscular transmission. Tetanic fade at 50, 100 and 200 hz did not change during the one hour period after infusion of morphine. Post-tetanic depression (PTD) of single twitch response increased progressively with time over the one-hour study period. This was most marked with the 100 and 200 hz tetanic frequencies. In contrast, control subjects displayed either no change, or a slight increase (post-tetanic facilitation; PTF), over a similar one-hour test period. We believe these results are best explained by a presynaptic action of morphine, whereby intraterminal mobilization of acetylcholine is impaired by the opiate narcotic drug.


Effects of lanthanum on the heart sarcolemmal ATPase and calcium binding activities. Effects of lanthanum on Ca2+-ATPase, Mg2+-ATPase, Na+-K+-ATPase, and calcium binding activities were studied in rat heart sarcolemma. Ten to 100 micrometers lanthanum depressed significantly the Ca2+-ATPase activity and 50--200 micrometers lanthanum inhibited the calcium binding activity. Lineweaver-Burk plots of the Ca2+-ATPase activity showed that the inhibition by lanthanum was competitive with calcium concentration. Neither Mg2+-ATPase nor Na+-K+-ATPase activities were affected by lanthanum when the assay medium contained 1 mM EDTA; however, in the absence of EDTA, these enzyme activities were significantly decreased by 10--100 micrometers lanthanum. Rat hearts perfused with HEPES buffer containing 0.5 mM lanthanum showed electron-dense deposits restricted to the outer cell surface and the sarcolemma obtained from these hearts also had the deposits, indicating that the membrane fraction isolated by the hypotonic shock--LiBr treatment method is of sarcolemmal origin. The Ca2+-ATPase activity of the sarcolemma isolated from lanthanum-perfused hearts, unlike the Mg2+-ATPase, Na+-K+-ATPase, and calcium binding activities, was significantly less than the control value. From these observations it is suggested that lanthanum may influence calcium movement across the sarcolemma by affecting sarcolemmal ATPase and calcium binding activities.


Effect of captopril (SQ 14225) on blood pressure, plasma renin activity and angiotensin I converting enzyme activity. Seven patients with essential hypertension and seven patients with hypertension associated with renal artery stenosis received captopril (SQ 14225), an inhibitor of angiotensin I converting enzyme. There was a significant reduction in mean blood pressure, from 176/113 +/- 4/3 mm Hg during the control period to 140/90 +/- 5/3 mm Hg during captopril administration. Five patients received captopril alone and nine patients needed hydrochlorothiazide in addition to control their blood pressure. Captopril produced a significant increase in peripheral plasma renin activity. When measured 12 hours after the administration of captopril the angiotensin I converting enzyme activity was found to be similar to that during the control period even though the blood pressure was at or near normal. These findings indicate that although captopril is an effective antihypertensive agent, its action does not depend only on inhibition of plasma angiotensin I converting enzyme activity.


Suspected reticuloendotheliosis in turkeys with cutaneous lesions reminiscent of fowl pox. Twelve turkeys from a flock of 2,500 had cutaneous lesions somewhat resembling fowl pox. Lymphoid-like tumors were evident in livers and spleens. Closer examination of cutaneous lesions, and histopathological examinations indicated skin lesions were also infiltrative and not unlike reticuloendotheliosis. Electron micrographs revealed the presence of C-type virus particles.


Differential protection of normal and malignant tissues against the cytotoxic effects of mechlorethamine. The radioprotective drug, WR-2721 (S,2-[3-aminopropylamino]ethyl-phosphorothioic acid), has been studied in terms of its ability to (a) protect mice against mechlorethamine (HN2)-induced hematopoietic death, and (b) alter the ability of HN2 injections to induce growth delay in a solid tumor, the Line 1 lung carcinoma. When WR-2721 was injected ip 15 minutes before iv injections of HN2, it increased resistance to hematopoietic death by a factor of 2, and the protection declined with a half-life of 1.5-2.0 hours. Similar administration of both drugs failed to alter the responsiveness of the Line 1 lung carcinoma to HN2-induced growth delays, except when the HN2 was given within 15 minutes after WR-2721. This interaction of the two drugs, when given within 5-15 minutes of each other, does not appear to be true protection at the tumor site, but rather appears to result from HN2 inactivation in the blood. When HN2 is given 30-60 minutes after WR-2721, it is possible to obtain a twofold increase in the tumor delay without risking increased hematopoietic injury.


Intra- and extracellular electrolytes and sarcolemmal ATPase in the failing heart due to pressure overload in dogs. An investigation of changes in the Mg2+ -dependent, Na+ -K+ -stimulated sarcolemmal ATPase and of intracellular electrolytes in the left failing heart due to pressure overload (aortic banding) was carried out in dogs. There was no change in the sarcolemmal Mg2+ -ATPase of the left or right ventricle for the whole duration (3 to 9 months) of left ventricular pressure overload. In the early phase (3 months) of aortic banding, when there was no haemodynamic evidence of left ventricular failure, there was also no significant change in the sarcolemmal Na+ -K+ -ATPase, extracellular space, or intra- and extracellular electrolytes. However, during 6 to 9 months of aortic binding when there was haemodynamic evidence of left ventricular failure (increased end-diastolic pressure, decreased cardiac index and (dP/dt)/IIP, enlarged heart), there was also a marked increase in the left ventricular sarcolemmal Na+ -K+ -ATPase and intracellular K+; and a decrease in the intracellular Na+ and Ca2+. The extracellular space in the left ventricle also increased significantly. Unlike the left ventricle, the right ventricle did not show any evidence of failure, not did it show any change in the sarcolemmal Na+ -K+ -ATPase and intracellular electrolytes during any period of aortic banding. These results suggest that the decrease in the myocardial contractility in failing heart due to pressure overload might be associated with a decrease in the intracellular Ca2+ as a result of an increase in the sarcolemmal Na+ -K+ -ATPase.


Morphological and physiological aspects of melanophores in primary culture from tadpoles of Xenopus laevis. Melanophores from tadpoles of Xenopus laevis (Daudin) were isolated by digestion of tail fins with acetyltrypsin and collagenase and maintained in primary culture for 6 weeks up to 3 months. Within 36 to 72 h the melanophores develop one to eight dendritic processes per cell; secondary and tertiary branchings of the processes were frequently observed. The melanophores in primary culture disperse under the influence of alpha-MSH or cyclic AMP; upon rinsing out these substances the cells aggregate. In darkness, about 40% of the cells disperse their pigment, whereas under illumination the pigment of the melanophores aggregates. To date, attempts to initiate cell division in melanophores have not been successful.


Cyclic GMP and cyclic AMP changes in response to folic acid pulses during cell development of Dictyostelium discoideum. Folic acid pulses induced developmental processes in agip 71, a morphogenetic mutant of Dictyostelium discoideum, strain Ax-2. Cells that had received folic acid pulses were able to form EDTA-stable cell aggregates and to complete full differentiation to fruiting bodies. In these cells no autonomous periodic activities were observed by light scattering. Folic acid pulses elicited increases in the concentrations of cyclic GMP and cyclic AMP. In undifferentiated cells, folic acid caused a rapid increase in the level of cyclic GMP without a significant change in the level of cyclic AMP. In an advanced developmental state folic acid caused an increase in cyclic AMP in addition to two successsive peaks of cyclic GMP. Experiments performed with the parent strain, Ax-2, also showed that during the development towards aggregation competence, cells acquired the ability to produce a cyclic AMP peak in response to folic acid.


Metabolic alterations of liver regeneration. XV. Cadmium-mediated depression of thymidine and thymidylate kinase induction in rats. Cadmium administered shortly before or after partial hepatectomy blocks in a dose-dependent manner the increase of thymidine and thymidylate kinase activities in regenerating rat livers. The effect of cadmium can be partially antagonized by simultaneous zinc administration. The intraperitoneal injection of cadmium is more effective than its subcutaneous administration. While there are in vitro differences in the sensitivity of thymidine kinase and thymidylate kinase towards Cd2+-, Zn2%- and Cu2+-ions, both enzymes are equally depressed following their in vivo administration. Cadmium displays the highest inhibitory activity and resembles in this respect beryllium [1].


Cyclic nucleotides levels in the liver of rats treated with CCl4. Treatment of rats with two different doses of CCl4 (respectively 2.5 and 0.5 ml/kg body wt. intragastrically) is followed by a rapid increase in the cAMP content of the liver. With 0.5 ml of CCl4, the increase occurs as early as 30 min after poisoning, namely about 4-5 h before the onset of triglyceride accumulation in the liver. The maximum increase has been at 6 h after administration of the hepatotoxin. In both experimental conditions, normal values are recovered only after 36-48 h. cGMP level appears unmodified during the whole observation period. Therefore the ratio cGMP/CAMP decreases consistently. The ATP level decreases significantly between 2 and 12 h. The increase in liver triglycerides level after CCl4 can be also a consequence of an impairment of microtubule function, leading to a decreased release of lipoprotein micelles from hepatocytes into the blood stream.


[Activation of the pituitary-adrenal axis by nipple suction in the lactating rat]. The response of the pituitary-adrenal axis to suckling in lactating Rats is dependent on two factors: 1. a psychic factor, which is identical by its effects to the psychological stress of the strange environment (transitory increase of plasma corticosterone and no significant modification of the content of ACTH in the two lobes of the pituitary); 2. a specific factor namely the stimulation of the nipples, which is distinguishable from the psychic factor by a longer release of corticosterone by the adrenals and a durable and significant decrease of the content of ACTH in the pars distalis.


[Role of tropomyosin in the sensitivity of (Na+ + K+) ATPase to ouabain]. EDTA treatment of isolated plasma membranes from MF2S cells increased 1,000 fold the sensitivity of (Na+ + K+) ATPase activity to ouabain. The original sensitivity of the enzyme to the drug is recovered after addition of tropomyosin together with Ca++ ions to the treated membranes.


[Molecular forms of human leukocytic alpha-galactosidase and N-acetyl-alpha-galactosaminidase]. 1. Normal human leukocytes present three molecular forms of alpha-galactosidase (EC 3.2.1.22) separated using electrofocusing: a new major form IV (pI 4.0) characteristic of leukocytes and two forms that exist in other tissues, form I (pI 4.5) and form II (pI 4.0). 2. Normal human leukocytes present only one molecular form (pI 4.5) of N-acetyl-alpha-galactosaminidase (EC 3.2.1.49) corresponding to alpha-galactosidase form II. 3. In leukocytes from patients with Fabry disease, the electrofocusing shows that the alpha-galactosidase lacking corresponds to forms I and IV, while the residual activity corresponds to form II (or N-acetyl-alpha-galactosaminidase).


[Choice of a spin label probe in order to study the recombination of beta-hydroxybutyrate dehydrogenase of a rat liver inner mitochondrial membrane with lecithin vesicles]. We find from studying the inhibitory effect of N-ethylmaleimide (NEM) on the enzymatic activity of beta-hydroxybutyrate dehydrogenase, that approximately one molecule of NEM is bound for one molecule of protein when the enzymatic activity is completely inhibited. Since the protein is a dimer this implies that each molecule of protein possesses only one thiol group in its catalytic center. Two long chain maleimide derivates: (10.3) NEM and (1.14) NEM conform, if a reasonable assumption is accepted to the conditions required for the study of the recombination of beta-hydroxybutyrate dehydrogenase with lecithin vesicles by spin label technique.


A prospective study of respiratory infection in adult asthmatics and their normal spouses. A prospective study of respiratory infections was performed in nineteen married asthmatics and their normal spouses who were examined at monthly intervals during a 1-year period. The colds described were associated with nasal symptoms, sore throat and usually malaise, fever, cough and hoarseness. The asthamtics reported a larger number of these symptomatic episodes than the non-asthmatics but significantly fewer of the episodes in the asthmatics were objectively confirmed by viral isolation or rise in serum titre of viral antibody. The frequency of respiratory infections was not influenced by the long term use of inhaled beclomethasone dipropionate and oral corticosteroid drugs. Less than 10% of the exacerbations of asthma were associated with respiratory infection. The disability resulting from respiratory infections in the asthmatics did not significantly exceed that in the non-asthmatics.


Liquid-chromatographic assay for urinary 3-methoxy-4-hydroxymandelic acid, with use of a periodate oxidative monitor. We describe a "high-performance" liquid-chromatographic procedure for measuring 3-methoxy-4-hydroxymandelic acid (vanillylmandelic acid) in urine. The compound is quantitatively extracted from urine by use of an automated system, including an anion-exchange resin. After elution from the resin and concentration by air drying, the urine extract is chromatographed on an octyl silica column, with use of a buffered mobile phase. To overcome interference from other organic acid in the extract, the column effluent is reacted with periodate in an analytical system to oxidize the analyte and internal standard to products that absorb at 360 nm. We evaluated the procedure for precision, linearity, analytical recovery, interfering substances, and correlation with an established procedure. The combination of a preliminary resin extraction, a liquid-chromatographic separation, and a periodate oxidative monitor results in a rapid, specific method that can be adapted for use in the clinical laboratory.


Comparison of current methods for high-density lipoprotein cholesterol quantitation. We compared six precipitation methods for high-density-lipoprotein cholesterol quantitation with an ultracentrifugation method, the accuracy of which was improved by correcting for 4% manipulative loss in the d greater than 1.063 fractions. For purposes of comparison, the apoprotein B-associated cholesterol (average 56 mg/L) measured by immunoassay in the d greater than 1.063 fractions was subtracted. In 65 plasma samples from men, women, and children, a heparin-Mn2+ procedure with Mn2+ at 46 mmol/L produced results slightly higher (+16 mg/L), while results with Mn2+ at 92 mmol/L averaged slightly lower (-8 mg/L) than the comparison ultracentrifuge method. Results that were about 5% low were obtained by the dextran sulfate 500-Mg2+ (-25 mg/L) and phosphotungstate-Mg2+ (-31 mg/L) methods. A heparin-Ca2+ method produced results 10% high (+58 mg/L). Results by a polyethylene glycol-6000 precipitation method were 12% low (-64 mg/L). Precision was better with the two heparin-Mn2+ and the dextran sulfate 500-Mg2+ procedures, with CVs of 4%, intermediate with phosphotungstate-Mg2+ and polyethylene glycol-6000 (CV 6-7%), and poorest with heparin-Ca2+ (CV 10%). Precipitation by phosphotungate-Mg2+ appeared more sensitive to reagent concentration and temperature variations than either the heparin-Mn2+ or dextran sulfate 500-Mg2+ methods. We conclude that these precipitation methods are not equivalent, but give rise to significant systematic differences in high-density-lipoprotein cholesterol quantitation.


A direct radio-immunoassay for plasma delta 4-androstenedione. Application to children. A RIA for the estimation of delta 4-androstenedione in human plasma without chromatographic purification is analyzed. Sensitivity, accuracy, precision and specificity are shown to be, at least, equivalent to those of methods involving chromatography.


A major locus for hyper-beta-lipoproteinemia with xanthomatosis. Complex segregation analysis of hyper-beta-lipoproteinemia with xanthomatosis has provided strong evidence for a major locus, in addition to significant polygenic effect and sibling environment. Estimates of gene frequency agree with values generally given in the literature.


Effect of veratrum alkaloids on right and left atrial receptors in the cat. 1. The effects of veriloid and veratridine on right and left atrial receptors have been studied in cats. 2. Threshold doses of these drugs for right atrial receptors were found to be almost twice those for left atrial receptors. Beyond threshold doses, however, the dose-response curves were similar. 3. The sensitivity of left atrial receptors to veratridine was not altered by an increase in PCO2 and/or decrease in PO2 of left atrial blood. Therefore, the difference in the behaviour of left and right atrial receptors to these drugs is not due to the differences in PO2 and PCO2 of blood in the two atria. They are more likely to be a feature of the regenerative  regions of the receptors.


Tumours at the spinocranial junction: some clinical and electromyographic aspects in relation to the symptomatology. On the basis of six cases of a tumour at the spinocranial junction, the clinical symptomatology is discussed with special reference to the occurrence of astereognosis and amyotrophy of the hand muscles. Possible mechanisms underlying these symptoms are discussed. In the authors' opinion, the frequent association of these symptoms is explained by the association of two pathogenic mechanisms, i.e., compression of the posterior column or interference with the arterial blood flow to the medial lemniscus, leading to astereognosis, and interference with the venous blood flow, leading to amyotrophy of the hand muscles.


Subacute combined degeneration of the spinal cord and the vitamin B12 metabolism, a clinical study. In 28 patients suffering from subacute combined degeneration of the spinal cord, vitamin B12 metabolism was investigated. Two postulates, proving vitamin B12 deficiency and excluding another cause for the clinical symptoms, have to be fulfilled. Two patients had no disturbance in their vitamin B12 metabolism. Seven patients had a distinct vitamin B12 deficiency. In the remaining 19 patients we found a mild vitamin B12 deficiency. Of these patients, 5 had had a subtotal gastrectomy, one had had a low absorption of vitamin B12, and 13 patients we could not find a distinct cause for the vitamin B12 deficiency. It is not impossible that nutritional habits can be hold responsible for this deficiency. The question whether these 13 patients should be treated with vitamin B12 for the rest of their lives is difficult to answer. It is a conditio sine qua non that in the patients with S.C.D. the vitamin B12 metabolism is examined circumstantially. By so doing, it may be possible to detect, in cases with minor clinical signs and symptoms of S.C.D., the cause of their illness.


Benign tumors of the cranial vault, a report of 12 cases. Twelve cases of different types of benign tumors of the cranial vault are described. In most cases there was a localised swelling which was sometimes painful. There were no neurological abnormalities due to the lesion. Radiological examination revealed a zone of radiolucency in all cases. In 8 patients the brain scan with 99mTc sodiumpertechnetate showed an increased uptake at the level of the bone lesion. Eight patients were operated upon and a total resection could be performed. In the four others only a biopsy was done. Histopathological examination revealed a fibrous dysplasia (3x), an eosinophilic granuloma (3x), an osteoid osteoma (2x), a cholesteatoma, aneurysmal bone cyst, an osteoclastoma and a fibroma. The pathological mechanisms underlying these different lesions are discussed.


Vertebral osteomyelitis and epidural abcess due to mucormycosis, a case report. A patient is described, with a history of pain in the neck, followed by a slowly progressive loss of muscle strength in both arms, followed by tetraplegia. Medical history included laryngectomy with partial hypopharyngectomy preceded by radiotherapy, because of carcinoma. X-ray of the cervical spine was suggestive for metastases. The patient died as a consequence of massive pulmonary embolism and at autopsy the cause of the neurological deficit turned out to be vertebral osteomyelitis and epidural abscess due to mucormycosis.


Vertebrobasilar insufficiency due to Tumoural emboli. A case, which is clinically characterized by a subacute vertebrobasilar syndrome, is presented. The necropsy reveals multiple infarcts in brainstem, cerebellum and left cerebral hemisphere, due to tumoural emboli of a non-detected primary tumour. It is postulated that the latter has to originate in the lungs in order to produce this unique type of cerebral arterial embolism.


Cerebrospinal fluid glutamine levels and EEG findings in patients with hepatic encephalopathy. The cerebrospinal fluid glutamine level was determined and an electro-encephalogram was made at roughly the same time in 41 patients on 55 occasions. A modified electro-encephalographic grading for practical use was introduced, because the classical electroencephalographic grading described by PARSONS-SMITH et al. (1957) does not apply to patients in deep coma. A significant correlation was found between the natural logarithms of cerebrospinal fluid glutamine levels and the electro-encephalic grade. In combination, these parameters provide valuable diagnostic and prognostic information in hepatic encephalopathy, and each can serve separately for follow-up purposes.


Grey scale ultrasound appearances in hepatocellular carcinoma. Thirty patients with histologically proven hepatocellular carcinoma were examined ultrasonically. All except two of these cases were reported ultrasonically as having a solid mass. Forty-three per cent of these lesions appeared multiple and 90% had irregular walls or boundaries. Stretching or distortion of the inferior margin of the liver is considered significant and 53% of cases demonstrated this feature. Just over two-thirds (67%) of the lesions were echogenic and under one-third (27%) were mixed lesions with echogenic and transonic areas. These transonic areas are considered to be due to the necrosis within the tumour. Most patients presented late and died within three months. Ninety per cent had clinical hepatomegaly or an epigastric mass. So far the main benefits of ultrasonography have been the recognition or exclusion of treatable disease such as liver abscess, cysts and congestive conditions of the liver, which may also present with unexplained hepatomegaly or epigastric masses. The possibility of a recognisable echo pattern for hepatocellular carcinoma emerged from this study. By conducting selective ultrasonic surveys in endemic areas and by employing the ultrasonic criteria described early diagnosis may be possible. Aspects of management and research may be assisted.


The CT scan and subtraction angiography in chemodectomas. The value of subtraction angiography and CT scanning in the diagnosis of chemodectomas are illustrated by two cases, a glomus jugulare tumour which exhibited true malignant behaviour, and a carotid body tumour. The characteristic early arterial blush of a chemodectoma, which can be obscured by bone, is clearly seen following subtraction techniques revealing tumours as small as 0.5 cm. The chemodectoma appeared as an isodense tumour on the CT scan with uniform contrast enhancement and well-defined margins. The appearances, however, are not diagnostic of a chemodectoma and may be simulated by other tumours.


Correlation between volumetric CT imaging and autopsy measurements of glioblastoma size. A three-dimensional reconstruction and display technique (THREAD SYSTEM) for serial computed tomography (CT) was employed in monitoring tumor volumes in two children under chemotherapy for glioblastoma multiforme of the cerebral hemispheres. Progressive diminution of tumor bulk was documented in the first patient and an increase in volume in the second. The first patient expired of the complications of his therapy and the second of transtentorial herniation. Independent measurements of the tumors as determined by a CT scan near the times of death and tumor dimensions measured at autopsy revealed good correlations between the radiographic and the anatomical data. The final CT measurement of tumor volume of 20 cm3 compared with an autopsy calculation of 13.3 cm3 in the first case. In the second case, CT volume was 417 cm3 and the actual volume of the primary tumor mass at autopsy was 437 cm3. The results suggest that the THREAD system is a practical method for monitoring the results of radiotherapy and chemotherapy in certain types of cerebral neoplasms.


Cadmium nephropathy.

Cadmium nephropathy. Cadmium is an inessential trace metal which accumulates in human tissues from contamination of food, water or air. The kidney is the critical organ following long-term, low-level absorption either by inhalation or ingestion; accumulation occurring in tubular epithelium in the form of a cadmium-metallothionein complex, giving rise to tubular dysfunction. In a group of 12 cadmium workers some of whom were followed for up to 16 years, tubular proteinuria, renal glycosuria, aminoaciduria, hypercalciuria and defects of concentration and acidification have been observed. Two men became recurrent renal stone formers and 1 man, who had nephrocalcinosis when first seen, later developed vitamin D-resistant osteomalacia. Renal tubular dysfunction following cadmium exposure may continue symptom-free for long intervals, but in a proportion of cases serious clinical effects may eventually develop.


Canine viral enteritis. I. Status report on corona- and parvo-like viral enteritides. This report attempts to provide the most recent information on salient features of two newly recognized viral enteritides of dogs. The viruses, a corona-like virus and canine parvo-like virus, are distinct but the disease manifestations in a particular outbreak often are similar. Laboratory study is therefore required. The parvoviral infection generally is more severe, with leukopenia (relative lymphopenia) a most prominent feature. Knowledge of the pathogenesis, epizootiology and the host's immune response is fragmentary at the present time, but research now in progress will doubtless expand knowledge of both diseases and of the viruses that cause them.


Fluorescence mapping of mitochondrial redox changes in heart and brain. Fluorescence techniques may be utilized to map changes in the distribution of mitochondrial redox states in heart and brain during ischemic or hypoxic stress. The basis of these techniques is the intrinsic fluorescence of reduced NADH and oxidized flavoprotein in mitochondria which respond to changes in critical oxygen supply. Ischemic areas in rabbit hearts induced by coronary ligation were detected and mapped based on the increase in NADH fluorescence in the ischemic zone. The width of the jeopardized normoxic tissue surrounding the ischemic area (less than 50--350 mu) was measured by combination of fluorescein angiography and NADH fluorescence. Areas of increased NADH fluorescence in gerbil brains after carotid artery ligation or induction of spreading depression were mapped in a similar manner. Intraoperative monitoring of flavoprotein fluorescence from human cerebral cortex after superficial temporal artery middle cerebral artery (STA-MCA) anastomoses demonstrated increased rates of cortical oxidative metabolism after the surgical procedures.


Oncornavirus modification of the Golgi apparatus. Two virus system, The Friend leukaemia virus (FLV) and the Rous sarcoma virus (RSV), were introduced into tissue both in vitro and in vivo. Both brought about substantial modification of the activity of the Golgi apparatus detectable as such by specific radioautographic studies. This modification was accompanied by changes in the development and social behaviour of the cells with some differences being detectable between the in vivo and in vitro studies.


Paracrystalline inclusions in various isolates of the blue-green bacteria Nostoc and Anabaena. A number of different crystalline inclusions were observed in various isolates of Anabaena and Nostoc. Membrane-limited crystalline bodies were observed in 7 of 20 isolates of Anabaena and 19 of 29 isolates of Nostoc. These are spherical, single membrane-limited bodies from 0.6 to 0.1 micron in diameter. In most of the isolates they contained needle-like crystals 20 A in thickness and up to 80 nm in length. In 9 of the isolates the inclusions contained granular and fibrillar material. The number of bodies per cell varied in the different isolates from only a few, observed in many sections, up to 5 in a single section of A. subtropica (B1618). Crystalloids were observed in the cytoplasm of Anabaena sp. (1551), N. calcicola (B382), Nostoc sp. (588), and N. punctiforme (1629). In Anabaena sp. (1551) the roughly cuboidal inclusions 0.6 micron in diameter were composed of 100 A thick osmiophilic striations spaced to produce a 150 A periodicity. In Nostoc sp. (588) the elongate, 0.1 micron by 2.5 micron, crystalloids were composed of 100 A thick osmiophilic striations spaced to produce a 200 A periodicity. N. punctiforme (1629) and N. calciola (B382) contained intrathylakoidal crystalloids which consisted of short curved segments with 100 A thick osmiophilic striations producing a 200 A periodicity. Granular areas were observed in 2 isolates of Anabaena and 5 of Nostoc. These bodies found in various locations in the cells, were interpreted to be elongate structures 0.2 micron thick, 1.2 micron long and about 5 micron in depth. These inclusions were composed of 15 nm diameter granules which in some section planes appeared in rows spaced 20 nm apart. Spherical bodies up to 0.7 micron in diameter and of medium electron density were observed in 4 isolates of Anabaena and 2 of Nostoc. Convoluted inclusions were found in N. calcicola (B382) and Anabaena sp. (1551). These roughly spherical bodies up to 0.8 micron in diameter contain lighter swirled areas.


Small cell carcinoma of the lung. Prognosis in relation to histologic subtype. Forty-six of 59 patients with small cell carcinoma of the lung who were treated with multiple drug chemotherapy and radiotherapy were subclassified according to the World Health Organization classification. Subtyping was not possible in the 13 other patients who were diagnosed on sputum cytology findings alone. There was no significant difference in extent of disease, response, duration of response to treatment, or median survival between the different subtypes. Two main difficulties arise in applying the subtyping classification. First, many tumors showed features of several histologic subtypes, implying the existence of a morphologic continuum within the general group of small cell anaplastic carcinomas. Second, tissue crushing artefact was common. Our results do not reveal any advantage in knowing the tumor subtype. It remains essential to differentiate small cell anaplastic carcinoma from other forms of lung carcinoma not responsive to chemotherapy.


Unilateral hyperlucent lung. Patent ductus arteriosus coexisting with bronchial carcinoid. We report an unusual case of patent ductus arteriosus (PDA) co-existing with bronchial carcinoid. The initial radiographic presentation was hyperlucent lung. Causes and possible mechanisms of unilateral hypoperfusion of the lung are reviewed briefly. Useful diagnostic modalities in evaluating a hyperlucent lung are also discussed.


Laparoscopic diagnosis of suspected liver neoplasms. Value of prior liver scans. To study the efficacy of standard blood tests of liver function (LFTs) and technetium-99m liver scan followed by laparoscopy in the diagnosis of solid malignant tumors of the liver, 100 consecutive patients were evaluated who had these tests because of clinical suspicion of liver neoplasm. Malignant liver tumors were present in 65 of the 100 patients. Laparoscopy diagnosed 60 of the 65, and the neoplasms were documented pathologically by directed biopsy (92% sensitivity, 100% specificity). In 8 patients, hepatoma was diagnosed and staged for possible resection. A negative "blind" biopsy had been performed in 23 of the 60 patients with liver neoplasms diagnosed at laparoscopy. LFTs and liver scans together were good screening tests (95% sensitivity) but had low specificity (46%). Liver scans also provided information as to probable tumor location in the liver, guiding the insertion site for the laparoscope, and directing deep-needle biopsies if no surface lesions were seen.


Effects of pancreatic polypeptide and vasoactive intestinal polypeptide on rat ileal and colonic water and electrolyte transport in vivo. Two gastrointestinal peptides, vasoactive intestinal polypeptide (VIP) and pancreatic polypeptide, suspected of being associated with symptoms of WDHA syndrome (pancreatic cholera) were tested on the rat small and large intestine for their effects on water and electrolyte transport. Intravenous infusion of VIP (14.3 microgram/kg/hr) inhibited net absorption of water and electrolytes in the ileum and reversed net absorption to net secretion in the colon. In contrast, bovine pancreatic polypeptide (52 microgram/kg/hr) did not inhibit absorption or stimulate secretion. These data indicate VIP causes colonic secretion in vivo, an effect previously shown only in vitro, and that bovine pancreatic polypeptide (at this dose) is not a secretagogue in the small or large intestine of the rat. Thus, while consistent with VIP being a contributory agent to the secretion of pancreatic cholera, the data do not support the notion that pancreatic polypeptide might be a causative agent in this syndrome.


[ADAM complex--maxillofacial abnormalities and abnormalities of the extremities caused by amniotic strangulations]. The anomalies of the ADAM complex arise through amniotic strangulations, adhesions and amputations. In the face cleft formations, displacements and deformities of various structures occur. The limbs exhibit constriction grooves, secondary syndactyle or amptuations. According to observations on 8 patients and information from the literature, the presentation of Adam complexes can vary to an extraordinary degree. The anomalies result from exogenous influences. In genetic counselling phenotypically similar congenital malformations must be separated.


alpha-Melanocyte-stimulating hormone and adrenocorticotropin in the regulation of glucocorticoid secretion during the perinatal period in sheep. To determine the role of other ACTH-like peptides in the regulation of glucocorticoid secretion in fetal sheep, we examined the responses of the adrenal gland of fetal and newborn sheep to comparable single doses of alpha MSH (75 micrograms) or ACTH (50 micrograms) during the last third of gestation and the first month of postnatal life. alpha MSH first increased the plasma glucocorticoid concentration at 121--130 days of gestation [from 16 +/- 1.5 to 36.9 +/- 9 (SE) ng/ml]; the response to alpha MSH persisted on days 131--140 of gestation and during the first month after birth. ACTH first increased the plasma glucocorticoid concentration at 131--140 days of gestation and increased it further in the first month after birth (from 18.9 +/- 3.6 to 97.0 +/- 10 ng/ml). The observations that the adrenal glands of fetuses and newborn lambs responded to alpha MSH at a dose comparable to that of ACTH and that the response to alpha MSH in the fetus preceded the response to ACTH may indicate that adrenal receptors mature during fetal development. These data also suggest that the regulation of the adrenal during fetal life may involve more than one tropic hormone.


Nitrogenous compounds reversibly inhibit the adenosine 3',5'-monophosphate response to thyrotropin: this effect is dissociated from altered guanine 3',5'-monophosphate levels. Nitrogenous compounds were studied for their effect on cGMP levels in the thyroid. Several agents, including sodium nitrite and hydroxylamine, could generate a rise of over 100-fold in cGMP. At a concentration of 20 mM, HN2OH and NaNO2 significantly reduced cAMP levels previously generated by 0.5 mU/ml TSH. If the slices were rinsed and placed in new buffer without the latter nitrogenous compound, the cAMP response to TSH was restored even though intracellular cGMP levels remained elevated. Thus, the counterregulatory effect of these agents on cAMP levels is not correlated with the presence of high levels of cGMP per se. Both the rise in cGMP and the fall in cAMP could still be detected when Ca++ was deleted from the incubation medium, in contrast to other reported stimuli that elevate cGMP levels in the thyroid.


Effect of stress, adrenocorticotropin or corticosteroid treatment, adrenalectomy, or hypophysectomy on hypothalamic immunoreactive adrenocorticotropin concentrations. Despite variations in plasma and/or pituitary ACTH concentrations, no changes in median eminence or medial basal hypothalamic immunoreactive ACTH-like concentrations were seen in rats studied 6 weeks posthypophysectomy, 12 h or 28 days after bilateral adrenalectomy, 6 days after implantation of corticosterone pellets (75 mg), after dexamethasone administration (300 micrograms/100 g BW, ip, twice daily for 4 days), or after chronic immobilization stress. The lack of concordant variation of brain, pituitary, and plasma ACTH concentrations further supports the suggestion of a nonpituitary, central nervous system origin of some portion of brain immunoreactive ACTH-like activity.


Gonadotropin binding and testicular function in old rats. Serum testosterone levels, testicular LH binding and the spermatogenic cycle were analyzed in rats 4 and 22 mo of age. With age, serum testosterone levels decreased from 3.2 to 0.63 ng/ml serum. There was no age related decline in testicular LH binding or changes in the spermatogenic cycle.


Biphasic inhibitory and stimulatory effects of thyrotropin on the mitotic activity of thyroid explants cultured in vitro. An influence of thyrotropin (TSH), dibutyryl cyclic AMP (dbcAMP), triiodothyronine (T3) and thyroxine (T4) on the mitotic incidence in the organ-cultured rat thyroid was studied. The study has revealed biphasic inhibitory and stimulatory effects of TSH on the thyroid mitotic activity. The inhibitory effect occurred 14 h after an exposure of the thyroid explants to TSH and was enhanced by T3. A similar inhibitory effect was observed after an exposure of the explants to dbcAMP. 24 h after the exposure of the thyroid explants to TSH a stimulatory effect on the mitotic incidence was observed. The latter effect was not reproduced by dbcAMP.


Purification and properties of sheep-liver aldehyde dehydrogenases. Sheep liver cytoplasmic aldehyde dehydrogenase was purified to homogeneity to give a sample with a specific activity of 380 nmol NADH min(-1) mg(-1). An amino acid analysis of the enzyme gave results similar to those reported for aldehyde dehydrogenases from other sources. The isoelectric point was at pH 5.25 and the enzyme contained no significant amounts of metal ions. On the binding of NADH to the enzyme there is a shift in absorption maximum of NADH to 344 nm, and a 5.6-fold enhancement of nucleotide fluorescence. The protein fluorescence (lambdaexcit = 290 nm, lambdaemisson = 340 nm) is quenched on the binding of NAD+ and NADH. The enhancement of nucleotide fluorescence on the binding of NADH has been utilised to determine the dissociation constant for the enzyme . NADH complex (Kd = 1.2 +/- 0.2 muM). A Hill plot of the data gave a straight line with a slope of 1.0 +/- 0.3 indicating the absence of co-operative effects. Ellman's reagent reacted only slowly with the enzyme but in the presence of sodium dodecylsulphate complete reaction occurred within a few minutes to an extent corresponding to 36 thiol groups/enzyme. Molecular weights were determined for both cytoplasmic and mitochondrial aldehyde dehydrogenases and were 212 000 +/- 8 000 and 205 000 respectively. Each enzyme consisted of four subunits with molecular weight of 53 000 +/- 2 000. Properties of the cytoplasmic and mitochondrial aldehyde dehydrogenases from sheep liver were compared with other mammalian liver aldehyde dehydrogenases.


Some comments on the article: 'The Structure of Human-Plasma Low-Density Lipoprotein B. An X-Ray Small-Angle Scattering Study' by K. Muller, P. Laggner, O. Glatter, and G. Kostner [Eur. J. Biochem. 82, 73--90 (1978)]. In their paper, K. Muller, P. Laggner, O. Glatter and G. Kostner report X-ray scattering and density experiments on human plasma low-density lipoprotein B, performed at different solvent densities (i.e. contrast variation method). The interpretation of the experimental data lead those authors to question the use of that method and particularly the use of the characteristic scattering functions in the study of serum lipoproteins. In the present paper it is shown that several aspects of their analysis are in fact highly questionable and that the objections raised are therefore unfounded.


Evidence for ligand-induced conformational changes in rabbit-muscle glyceraldehyde-3-phosphate dehydrogenase. The tetrameric glyceraldehyde-3-phosphate dehydrogenase from rabbit muscle binds NAD+ and some of its analogues in a negatively cooperative manner, whereas other NAD+ analogues bind non-cooperatively to this enzyme. Subsequent to alkylation of a fraction of the active sites of the enzyme with the fluorescent SH reagent N-iodoacetyl-N'-(5-sulfo-1-naphthyl)-ethylenediamine, it was found that the alkylated sites bind NAD+ and NAD+ analogues with a markedly reduced affinity as compared with non-alkylated sites. It was therefore feasible to measure the fluorescence and the circular polarization of the luminescence of the enzyme-bound alkyl groups as a function of binding of NAD+ and of NAD+ analogues to the non-alkylated sites. The changes observed indicate that ligand binding to the non-alkylated sites induces changes in the fluorescence properties of the alkyl groups bound to neighbouring subunits, most likely through the protein moiety. The nature of these changes appears to depend on the structure of the coenzyme analogue. The binding of the non-cooperative binders acetyl-pyridine--adenine dinucleotide, ATP and ADP-ribose induce different conformational changes in the neighbouring vacant subunit, as monitored by the spectroscopic properties of the bound alkyl group. These results in conjunction with other data support the view that the negative cooperativity in NAD+ binding to glyceraldehyde-3-phosphate dehydrogenase results from ligand-induced conformational changes. Furthermore, these results further support the view that subtle structural changes in the coenzyme molecule determine the nature of the conformational changes induced within the enzyme tetramer.


Aggregation of sponge cells. Isolation and characterization of an inhibitor of aggregation receptor from the cell surface. From the cell membranes of the sponge Geodia cydonium a component was isolated and purified which inhibits the aggregation factor isolated from the same source; the component was termed anti-aggregation receptor. This molecule was characterized as a glycoprotein (54% neutral carbohydrate) and its molecular weight is in the range of 180,000 One biological site of the anti-aggregation receptor was determined to be D-galactose. Indirect evidence presented seems to indicate that this molecule is present in an active form in aggregation-deficient cells and absent in aggregation-susceptible cells.


Studies on the methylation of cytoplasmic ribosomal RNA from cultured higher plant cells. The methylation of cytoplasmic ribosomal RNA of cultured sycamore cells (Acer pseudoplatanus L.) was investigated. Labelled 17-S and 26-S rRNA were prepared from cells that had been incubated with either [32P]phosphate, [Me-3H]methionine or [Me-14C]methionine. Ion-exchange resin chromatography of 0.3 M KOH or 1 M HCl hydrolysates and two-dimensional chromatographic analyses of phosphodiesterase plus phosphatase digests of 17-S and 26-S rRNA were performed. 17-S and 26-S rRNA contain 49 and 91 methyl groups per molecule, respectively. These values were verified in sevemral ways. The high degree of methylation of sycamore rRNA, particularly for the 26-S rRNA, contrasts with the situation in all other investigated organisms. Several methylated bases were identified. 7-Methylguanine and 5-methylcytosine both occur in 17-S and 26-S rRNA. N6-Methyladenine and N6,N6-dimethyladenine are restricted to the 17-S rRNA while 3-methyluracil and 1-methyladenine occur in the 26-S rRNA. One hypermodified uridine was also tentatively identified in the small rRNA. In 17-S rRNA, there is one copy of 7-methylguanine, N6-methyladenine and hypermodified uridine and two copies of N6,N6-dimethyladenine. 3-Methyluracil, 1-methyladenine and 5-methylcytosine occur twice, twice and three times, respectively, in 26-S rRNA. 7-Methylguanine and 5-methylcytosine are only in submolar amounts in the 26-S and 17-S rRNA, respectively. There are 40 +/- 2 and 83 +/- 3 2'-O-methylriboses per 17-S and 26-S rRNA molecule, respectively. In addition to the four 2'-O-methylnucleosides, one 2'-O-methylpseudouridine is present in the 17-S rRNA. Several lines of evidence argues for a non-random distribution of the methylriboses. In particular, one and seven Nm-Nm-Np structures occur in the 17-S and 26-S rRNA, respectively. The data are discussed comparatively with the methylation pattern of Escherichia coli, yeast and HeLa cell rRNA.


Properties of herpes simplex virus DNA polymerase and characterization of its associated exonuclease activity. Herpes simplex virus (HSV) DNA polymerase was isolated on a large-scale from African green monkey kidney cells infected with HSV type 1 (HSV-1) strain Angelotti. After DNA-cellulose chromatography the enzyme showed a specific activity of 48,000 units/mg protein. Three major single polypeptides with molecular weights of 144,000, 74,000 and 29,000 were copurified with the enzyme activity at the DNA-cellulose ste. By its chromatographic behavior and by template studies, the HSV DNA polymerase activity was clearly distinguishable from cellular alpha, beta and gamma DNA polymerase activities. Two exonucleolytic activities were found in the DNA-cellulose enzyme preparation. The main exonucleolytic activity, which degraded both single-stranded and double-stranded DNA to deoxynucleoside 5'-monophosphates, was separated by subsequent velocity sedimentation. The remaining exonucleolytic activity was not separable from the HSV DNA polymerase by several chromatographic steps and by velocity sedimentation at high ionic strength. This novel exonuclease and HSV DNA polymerase were equally sensitive both to phosphonoacetic acid and Zn2+ ions, inhibitors of the viral polymerase. Similar to the 3'-to-5'-exonuclease of procaryotic DNA polymerases and mammalian DNA polymerase delta, the HSV-polymerase-associated exonuclease catalyzed the removal of 3'-terminal nucleotides from the primer/template as well as the template-dependent conversion of deoxynucleoside triphosphates to monophosphates.


Cytochrome c oxidase subunits in nuclear and extranuclear cytochrome-aa3-deficient mutants of Neurospora crassa. The mitochondria of cytochrome-aa3-deficient Neurospora crassa mutants were screened for the seven polypeptide constiuents of cytochrome c oxidase. The polypeptides of the holoenzyme and the unassembled or partially assembled subunits were detected by sodium dodecyl sulfate/acrylamide gel electrophoresis of immunoprecipitates obtained with antiserum to the holoenzyme as well as to several individual subunits. With respect to the mitochondrially synthesized polypeptides of the oxidase, subunits 1 to 3, the results obtained from the analysis of immunoprecipitates were confirmed through the direct electrophoretic analysis of mitochondrial translation products. The results were as follows. 1. The mitochondria of the cya-2-8 and cya-3-16 nuclear mutants and the [exn-5] cytoplasmic mutant contained a protein complex immunoprecipitated by anti-holoenzyme antibody and composed of the complete set of the seven cytochrome oxidase polypeptides. Only the oxidase subunits 5 and 6 were immunoprecipitated by anti-holoenzyme antibody from the mitochondria of the cyt-2-1 and 299-1 nuclear mutants, even though at least some of the mitochondrially synthesized polypeptides were detected in both mutants by subunit specific immunoprecipitation. 2. A 'subunit 1' polypeptide larger than the authentic subunit-1 polypeptide of wild-type cytochrome oxidase was found in the mitochondria from two nuclear mutants, cyt-2-1, and 299-1 and the [mi-3] cytoplasmic mutant. This larger polypeptide may be an unprocessed precursor of the 'mature' subunit 1 protein of the holoenzyme. No changes in the apparent molecular weights were found for the polypeptide subunits of cytochrome oxidase in mitochondria of the [exn-5] cytoplasmic mutant and the cya-2-8 and cya-4-23 nuclear mutants. 3. A nuclear mutant, 299-1, lacks the mitochondrially synthesized subunit-2 polypeptide of cytochrome oxidase. When cells were labelled in the presence of cycloheximide, the subunit 2 content of mitochondria from mutants [exn-5], cya-2-8, cya-3-16 and cya-4-23 was lower than in mitochondria from wild-type. This deficiency, however, does not appear to be sufficiently severe to fully account for the lack of cytochrome aa3 in these mutants. The cya-4-23 nuclear mutant either is severely deficient in or lacks cytochrome oxidase subunits 5 and 6. On the basis of these and previously reported observations, it is proposed that the cytochrome oxidase deficiencies of as many as seven of the eight N. crassa cytochrome-aa3-deficient mutants could be caused by genetically imposed alterations in regulatory systems controlling the production of different components of the enzyme.


Regulation of protein synthesis.

Regulation of protein synthesis. A system of translational control in eukaryotes consists of (a) a proinhibitor and (b) an inhibitor of polypeptide chain initiation. The inhibitor (active eIF-2 kinase), a cAMP-independent protein kinase, catalyzes the phosphorylation by ATP of the small subunit of the polypeptide chain initiation factor eIF-2. This blocks the interaction of eIF-2 with eIF-2 stimulating protein (ESP) without which eIF-2 is unable to form an initiation complex, a prerequisite for translation. Our observations are consistent with the view that the proinhibitor (inactive eIF-2 kinase) is converted to the inhibitor by phosphorylation catalyzed by a cAMP-dependent protein kinase. This is analogous to the conversion of inactive phosphorylase kinase to active phosphorylase kinase. As in the case of phosphorylase kinase and phosphorylase, the modification of activity produced by phosphorylation of eIF-2 kinase and eIF-2 itself is probably reversed by dephosphorylation catalyzed by specific protein phosphatases (see diagram in Fig. 12) but no evidence bearing on this aspect of the problem is yet available. Hemin inhibits the cAMP-induced dissociation of the regulatory and catalytic subunits of cAMP-dependent protein kinase by binding to the regulatory subunit of the enzyme and blocking, through an allosteric effect, the binding of cAMP. Thus, hemin prevents the activation of eIF-2 kinase by inhibiting the cAMP-dependent protein kinase.


The effect of prednisolone on neuromuscular transmission in the rat diaphragm. Prednisolone, in concentrations of 0.004--0.032 mmol/l, increased the amplitude of the miniature end-plate potentials (MEPPs). A maximum increase to 134% of the control values was seen at 0.016 mmol/l. At higher prednisolone concentrations the MEPP amplitude gradually decreased to reach 77% of the control value at 0.62 mmol/l. The MEPP frequency was increased to twice the control value at 0.62 mmol/l. The quantal content of the end-plate potential (EPP), however, was not influenced by prednisolone. The response to iontophoretically applied acetylcholine was diminished, especially at 0.62 mmol/l prednisolone. Prednisolone, therefore, caused presynaptic effects as was shown by an increase in unitary MEPP amplitude and by a considerable number of giant MEPPs, which at increasing prednisolone concentrations was antagonized increasingly by a postsynaptic depressant effect. These results also provide an explanation for the adverse effects of prednisolone on the end-plate potential and on neuromuscular transmission.


Opiate antagonist--receptor interaction unchanged by acute or chronic opiate treatment. The apparent pA2 of naloxone, a measure of the apparent receptor affinity for the antagonist, was estimated in naive rats, in rats after acute and various schedules of chronic morphine pretreatment, and in rats during abrupt withdrawal. The results were adjusted to compensate for the amount of pretreatment drug in the rat brains as, otherwise, such a drug could have inflated the apparent pA2 values (see appendix). After this adjustment, a tendency of the apparent pA2 values to be increased after chronic pretreatment disappeared. The apparent pA2 of naloxone in acutely pretreated rats was also unchanged from that in naive rats, while the results in abruptly withdrawn rats were ambiguous. Thus, no change in the affinity of the antagonist for opiate receptors was found and theories suggesting that a conformational change takes place in the opiate receptor during the development of tolerance/dependence could not be supported.


Effects of external calcium ions and strontium ions on interactions of isoprenaline and its competitive inhibitor with beta-adrenoceptors in the intestinal smooth muscle. The effects were tested of Ca ions and Sr ions on interactions of isoprenaline and its competitive inhibitor with beta-adrenoceptors in the taenia caecum from the guinea pig. Ca ions were involved in the combination of isoprenaline with the beta-adrenoceptors but not involved in the combination of the competitive inhibitor with the beta-adrenoceptors. Sr ions could not substitute for Ca ions in the isoprenaline--receptor interaction.


Modulation of the evoked release of noradrenaline in canine saphenous vein via presynaptic receptors for adenosine but not ATP. The effects of adenosine, ATP and adenosine beta, gamma-methylene-5'-triphosphate (APPCP) were compared in the saphenous vein of the dog. Adenosine and ATP in concentrations which did not affect the response to exogenous noradrenaline, depressed that to nerve stimulation; this effect was antagonized by theophylline. APPCP increased tension in unstimulated veins and in those made to contract with noradrenaline and nerve stimulation. The results suggest that the inhibitory effect of ATP on adrenergic neurotransmission is due to its rapid breakdown to adenosine, which acts on prejunctional P1 purinoceptors.


Effect of an angiotensin converting enzyme inhibitor (SQ 14,225) on beta-adrenergic and angiotensin-induced thirsts. The effect of acute administration of SQ 14,225, a new angiotensin converting enzyme inhibitor, on the drinking response of female rats administered either isoprenaline, angiotensin I, or angiotensin II was studied during 2 h after treatment. Administration of isoprenaline (25 micrograms/kg body wt) was accompanied by a significant increase in water intake when compared with saline-treated controls. Acute administration of a constant dose of isoprenaline (25 micrograms/kg body wt) and increasing doses of SQ 14,225 (5--50 mg/kg) was accompanied by a dose-related, linear decrease in water intake. Acute administration of either angiotensin I or angiotensin II (200 micrograms/kg body wt) was accompanied by a significant increase in water intake. The dipsogenic response to angiotensin II was not affected by acute administration of 35 mg SQ 14,225/kg body wt. However, at the same dose of SQ 14,225, angiotensin I-induced thirst was attenuated. Since isoprenaline-induced and angiotensin I-induced, but not angiotensin II-induced, thirsts are blocked by SQ 14,225, the results suggest that isoprenaline-induced thirst is mediated by way of the renin--angiotensin system.


Changes of adrenoceptor-mediated responses in the pithed rat during propylthiouracil-induced hypothyroidism. Adrenoceptor-mediated responses of the cardiovascular system during propylthiouracil-induced hypothyroidism were investigated. Interfering nervous reflexes could be circumvented by using pithed rats. Cardiac beta-adrenoceptor-mediated acceleration of heart rate following electrical stimulation or injection of noradrenaline and isoprenaline was markedly diminished even after 2 weeks of treatment. This loss of beta-sensitivity prevented the study of possible changes of presynaptic regulatory adrenoceptors in this test model. Taking the increase in diastolic blood pressure after application of alpha-agonists as index of the sensitivity of vascular alpha-adrenoceptors we found these to have been desensitized in the hypothyroid state. According to these results the lack of thyroid hormones exerted a similar effect on cardiac beta- and on vascular alpha-adrenoceptors.


Adrenoceptors involved in the contractile activity of islated pregnant rat uterus. Cumulative log dose-response curves of soterenol, isoproterenol, phenylephrine and norepinephrine on isolated pregnant rat uterus at different days of gestation, were investigated. Soterenol produced a sustained inhibition of spontaneous motility during the whole pregnancy and this effect was blocked by butoxamine. The affinity of myometrium for the beta 2-adrenoceptor agonist was parallel to the concentration of progesterone in plasma during pregnancy. Isoproterenol, norepinephrine and phenylephrine caused dual, alpha- and beta-mediated responses, their relative dominance varied with the concentration and the days of pregnancy, alpha-Adrenoceptor effects of the amines coincided with increased plasma concentrations of estrogens, whereas beta ones were in parallel with the increment of plasma progesterone. It is concluded that: (a) there exist in the pregnant rat uterus beta 2-receptor-mediated responses influenced by the length of gestation; and (b) the concentration-pregnancy-dependent biphasic actions of isoproterenol, norepinephrine or phenylephrine suggest that their variable hormone-modulated ability interacts with both alpha- and beta-adrenoceptive uterine sites.


The establishment of the suspension Epstein-Barr virus producing cell lines from patients with tumoral diseases. Isolation of the Epstein-Barr virus (EBV) in suspension lymphoblastoid cell lines from human patients with tumor diseases, mainly malignant lymphoma, has been described. It has been shown that the EBV was isolated from human patients with myeloid type of leukemia in 75% of cases. A similar virus was also isolated from patients with Hodgkin's disease and leukemoid reaction of the myeloid type for lung cancer. Morphological, cytochemical, immunological, and cytogenetic characteristics of the cell lines in which the EBV is replicated have been investigated.


Amphiphilic spectroscopic probes utilizing metal chelates. The synthesis and initial applications are reported for 1-[p-(palmitamido)-phenyl]ethylenedinitrilotetraacetic acid. The results demonstrate the versatility of this spectroscopic probe molecule, which allows choice of a particular technique for a particular system as well as use of multiple spectroscopic techniques for complementary information about hydrophobic regions in biological systems.


The inability of nuclear dehydrogenating clostridia to oxidize bile salt hydroxyl groups. In a survey of the intracellular bile salt oxidoreductase activity in fecal bacteria, 16 strains of nuclear dehydrogenating clostridia and 2 strains of non-nuclear dehydrogenating C. paraputrificum were demonstrated unable to oxidize cholate at any of the 3 OH groups. Since nuclear dehydrogenation at the delta-1 and delta-4 position requires a 3-oxo precursor steroid, it appears that these organisms require the presence of a 3 alpha-hydroxysteroid dehydrogenating organism for nuclear dehydrogenation.


Increase in Na+, K+ -ATPase enzyme units in liver and kidneys from essential fatty acid deficient rats. (3H)-Ouabain binding to liver and kidney preparations was utilized to estimate the number of Na+, K+-ATPase enzyme units in livers and kidneys from rats fed 2% corn oil supplemented or fat-free diets. The specific (3H)-ouabain binding in liver and kidney preparations from fatty acid deficient rats was increased approximately 40%, but the affinity of the binding sites for ouabain (Kd-value) remained unchanged. The increased concentration of Na+, K+-ATPase enzyme units observed in the essential fatty acid deficient rats may contribute to the reduced body fat accumulation and elevated heat production observed in these animals.


Effect of acetone on VLDL secretion by the isolated rat liver. High acetone levels occur in uncontrolled diabetes and after isopropanol administration to rats. In both conditions, very low density lipoproteins (VLDL) secretion is depressed. Acetone, hoever, failed to affect the VLDL secretion rate by the isolated perfused rat liver, suggesting that this metabolite is not involved in impaired VLDL production in diabetes and after isopropanol administration.


Absence of dopamine sensitive adenylate cyclase in the A10 region, the origin of mesolimbic dopamine neurones. Dopamine (DA) failed to stimulate the adenylate cyclase of the mesolimbic A10 DA nerve cell body area, in contrast to tis activating effect in the nigrostriatal A9 DA cell body area. The enzyme was stimulated by GMPPNP (a GTP analog) and NaF. This indicates the absence in the A 10 cell area of DA receptors with functional coupling on adenylate cyclase, in contrast to the A9 cell area where such DA receptors are believed to be located on afferent axon terminals.


Inverse regulation of spore germination and growth by cyclic AMP in Streptomyces hygroscopicus. The cyclic AMP level in germinating spores of Streptomyces hygroscopicus rises to a miximum ag outgrowth of germ tubes. Exogenous cyclic AMP results in an inverse effect on germination speed and growth.


Activation of the electrogenic Na-pump of cardiac muscle fibres by ACh in K-free solutions. The ionic mechanism of the membrane outward current (ACh-current) of bullfrog atrium muscle, induced by acetylcholine in K-free solution, was analyzed by a voltage-clamp experiment. The results suggested that the ACh-current was induced not only by an increase in K-conductance but also by an activation of the electrogenic Na-pump.


Depression of spontaneous and ionophore-induced neurotransmitter release by Salmonella. Exposure of frog neuromuscular junctions to heat-killed, lyophilized Salmonella typhimurium (SR 11) produces an early increase in spontaneous transmitter release followed by depression of release and blockade of the obligatory release usually induced by ionophore X537A.


On the action of vitamin C on the neuromuscular junctions. High concentrations of ascorbic acid produce complete blockage of the transmission of nervous stimuli on the isolated neuromuscular preparation of phrenic nerve-hemidiaphragm of the rat.


Spermatogenesis in men exposed to polybrominated biphenyl (PBB). Experience with other halogenated hydrocarbons, animal experiments, and individual complaints indicated that polybrominated biphenyl (PBB) might have an effect on human spermatogenesis. Analysis of semen from 52 PBB-exposed men as compared with analysis of semen from a control group of 52 men not exposed to PBB revealed no differences in the distribution of sperm counts, motility, or morphology.


Initiation of hamster sperm motility from quiescence: effect of conditions upon flagellation and respiration. The point within the male reproductive tract where sperm motility originates varies with mammalian species. Premotile sperm from hamster, a species whose sperm are still quiescent in the epididymis, were used here to investigate further the parameters involved in the initiation of sperm motility. Two types of motility were produced: (1) partial, weak flagellation by simple dilution; (2) strong, complete motility by inducers (calcium, cyclic adenosine 3':5'-monophosphate) in the presence of amplifiers (caffeine, spermine). Expansion of initiation conditions to test tube volumes revealed that, at high sperm dilutions, bicarbonate could also induce motility. The respiratory consequences of sperm motility induction were measured. A Large, short-term burst of oxygen consumption occurred at a time paralleling the previously reported shifts in nucleotide levels associated with this event.


The effect of sulfhydryl and amino group reagents on human lymphocyte--sheep erythrocyte rosettes. We have studied the influence of certain chemical groups, located on the surface of lymphocytes or sheep red blood cells, on the ability of these cells to form spontaneous rosettes. We found that fluorescamine, which reacts with amino groups, inhibits rosette formation, while p-chloromercuriphenylsulfonic acid, which binds to sulfydryl groups, increases the percentage of the rosette-forming lymphocytes. The possible mechanism of action of the above reagents is discussed.


Cytologic uniformity of breast cancer from different localizations: a pattern analysis study. Using cytologic pattern analysis it is shown that regions of different histologic structure in one breast cancer are of cytologic homogeneity. The so-called PFORM -- the quotient of the total outline length of DNA-particles to the nuclear area -- gives an objective cytologic parameter which is independent of DNA content of cancer cells. This objective parameter together with the subjective cytologic parameters described earlier may be a better basis for describing the biologic behavior of breast cancer than histologic structure which is inhomogeneous very often in a lump.


Filipin prevents and reverses insulin stimulation of rat adipocyte phosphodiesterase. A membrane fraction prepared from isolated rat adipocytes contained an insulin-sensitive cyclic nucleotide phosphodiesterase (EC 3.1.4.17) which catalyzed the hydrolysis of both adenosine 3',5'-monophosphate (cAMP) and guanosine 3',5'-monophosphate (cGMP). The rate of hydrolysis of cGMP was about one-third that of cAMP. The hydrolysis of the two nucleotides appeared to be assoicated with one catalytic site: one nucleotide interfered with the hydrolysis of the other, in a manner predictable from the kinetic constants in that the Km of one nucleotide as a substrate was comparable to its Ki as an inhibitor of the hydrolysis of the other nucleotide. Incubation of the adipocytes with insulin increased the Vmax of phosphodiesterase without affecting the Km values for either substrate. After adipocytes had been treated with filipin, a membrane perturbant, at a concentration that did not cause cell lysis, the response of phosphodiesterase to insulin was obliterated. Further, the insulin-stimulated phosphodiesterase activity was reversed when hormone-treated cells were subsequently incubated with this agent. These results suggest that the response of membrane phosphodiesterase to insulin is impaired once adipocytes have been exposed to filipin, either preceding or following the incubation with insulin.


Inhibition by estradiol-17 beta of porcine thecal androgen production in vitro. Thecal preparations from medium-sized procine ovarian follicles (3.5-5 mm diameter) were incubated for 4 h in a chemically defined medium in the presence or absence of highly purified luteinizing hormone (LH) and/or estradiol-17 beta (estradiol). LH (1 microgram/ml) stimulated the thecal production of testosterone (T) and dihydrotestosterone (DHT) by 2- to 3-fold. Although estradiol (10 microgram/ml) alone had only a slight but non-significant inhibitory effect on basal testosterone production, it significantly inhibited the  production of both T and DHT as well as decreasing the DHT/T ratio in a dose-related manner in the presence of LH. Production of cyclic adenosine 3',5'-monophosphate (cAMP) and progesterone by the thecal cells was stimulated 50-to 200-fold and 2.5-fold, respectively, by LH. Estradiol had no significant effect on thecal cAMP and progesterone production in the presence or absence of the gonadotropin. These findings are consistent with the concept that estradiol produced by granulosa cells following hormonal stimulation may serve as a local negative feedback mechanism to control thecal androgen production.


Hormonal regulation of thyroidal protein phosphokinase activities. II. Differential sensitivity of type I and type II cAMP-dependent enzymes to the treatment of rats with thyroxine. Two cAMP-dependent protein kinase activities can be separated from rat thyroid cytosol. Their elution properties during the gel chromatography, as well as their sedimentation coefficients after sucrose-gradient ultracentrifugation and their capacity to dissociate in the presence of histones, suggest they are like the Type I and Type II protein kinases described in many other tissues. The sensitivity of the two types of kinases to thyroxine treatment is different. The activity of Type I is not changed during the first 5 days of treatment. Thereafter, it decreased by about 50% and is maintained at that level for up to one month. The activity of the Type II enzyme decreased rapidly by about 30-40% already on the second day of treatment, and after 10 days it reached 50% of the initial level. This differential reactivity of the two types of enzymes to the thyroxine treatment leads to profound modifications in their relative activities between the second and the fifth day of treatment. The significance of these results and the possible role of the two types of kinases in the control of different steps of iodine metabolism have been discussed.


Evidence for the involvement of lutropin-independent RNA synthesis in Leydig cell steroidogenesis. The effect of incubating purified Leydig cells in Eagle's medium and the subsequent effect of the RNA synthesis inhibitors, actinomycin D and cordycepin, on lutropin-stimulated testosterone synthesis have been investigated. The inhibiting effect was found to be inversely related to the time of preincubation; with cells preincubated for 0, 1, 2 and 3 h with Eagle's medium only, followed by 2-h incubation with lutropin with and without actinomycin D, testosterone synthesis was inhibited by 37 +/- 4, 31 +/- 3, 18 +/- 4 and 14 +/- 3% respectively (means +/- s.e.m., n = 5). In cells that had been preincubated for 3 h there was no significant effect of actinomycin D on testosterone synthesis during the first hour of incubation with lutropin. Thereafter the inhibition increased with time reaching a maximum of 30% after 5 h. The effects of preincubation were not due to endogenous lutropin in the Leydig cells because cells isolated from hypophysectomized rats gave similar results. The inhibition of [3H]uridine incorporation into the Leydig cell RNA was 80 +/- 1% with 8 microgram/ml actinomycin D. Increasing the concentration of this inhibitor to 80 microgram/ml did not significantly increase the inhibition of [3H]uridine incorporation or lutropin-stimulated steroidogenesis in preincubated and non-preincubated cells. With cordycepin the inhibition of both RNA synthesis and lutropin-stimulated testosterone synthesis in non-preincubated cells were the same; with 25.1--251 microgram/ml approx. 30--70% resp. With preincubated cells (3 h), 0--50% inhibition of testosterone synthesis was obtained respectively. The inhibitory effect of actinomycin D oimilar to that obtained with lutropin. These observations suggest that during preincubation and independently of lutropin, synthesis of intermediates, including RNAs required for stimulation of steroidogenesis, takes place and that subsequent stimulation of steroidogenesis by lutropin occurs without further de novo RNA synthesis. These results provide evidence for a permissive role of specific RNA and protein synthesis in the action of lutropin on testosterone synthesis in the Leydig cell.