Parameters of the three-pool model of the turnover of plasma cholesterol in normal and hyperlipidemic humans. Long-term studies (32-49 wk) of the turnover of plasma cholesterol were conducted in 24 subjects. Eight subjects were normilipidemic, six had hypercholesterolemia, eight had hypercholesterolemia and hypertriglyceridemia, and two had hypertriglyceridemia alone. 10 of the hyperlipidemic patients had a definite familial disorder. In all subjects (except one for whom complete data were not available), the same three-pool model previously described gave the best fit for the data. The parameters of the three-pool model observed in the normal subjects were compared with the model parameters found in the patients with the different kinds of hyperlipidemia. In addition, single and multiple regression analyses were conducted to explore the relationships between the model parameters and various physiological variables, including age, body size, and serum lipid concentrations. Using this approach, significant differences between groups, or correlations with serum lipid levels were seen for several parameters of the three-pool model: the production rate (PR); the size of the rapidly exchanging pool 1 (M1); all estimates of the size of the most slowly equilibrating pool 3 (M3); and the rate constant k21. The PR in normal subjects (1.14 +/- 0.19 g/day, mean +/- SD) was not significantly different from that found in patients with hypercholesterolemia, with or without hypertriglyceridemia. The major determinant of cholesterol PR was overall body size, expressed either as total body weight or as surface area. The correlations between PR and indices of adiposity (percent ideal weight and excess weight), although statistically significant, were much weaker in this nonobese population. After adjustment for body size variation, cholesterol PR was not correlated with the serum cholesterol concentration but was probably (P less than 0.05) correlated with the triglyceride concentration. When the two patients with very high triglyceride concentrations were excluded, however, no correlation was observed between adjusted PR and triglyceride level. It is probable that hypertriglyceridemic patients represent a heterogeneous population, in which the majority do not show increased cholesterol PR. M1 was correlated with all body size variables, but most strongly with excess weight. After adjusting for the effects of body size, M1 was also correlated and triglyceride. Major differences were found in the relationships between the physiological variables and the sizes of pools 2 and 3. M2 was correlated neither with any of the indices of body size or adiposity, nor with the serum levels of either cholesterol or triglyceride. In contrast, all estimates of M3 were correlated with indices of adiposity (but not of overall body size) and with the serum cholesterol concentration. Thus, the amount of cholesterol in slowly equilibrating tissue sites appears to particularly increase with elevations of the serum cholesterol level. The results also confirm previous data that adipose tissue cholesterol is an important part of pool 3.


Identification of beta-adrenergic receptors in human lymphocytes by (-) (3H) alprenolol binding. Human lymphocytes are known to posessess a catecholamine-responsive adenylate cyclase which has typical beta-adrenergic specificity. To identify directly and to quantitate these beta-adenergic receptors in human lymphocytes, (-) [3H] alprenolol, a potent beta-adrenergic antagonist, was used to label binding sites in homogenates of human mononuclear leukocytes. Binding of (-) [3H] alprenolol to these sites demonstrated the kinetics, affinity, and stereospecificity expected of binding to adenylate cyclase-coupled beta-adrenergic receptors. Binding was rapid (t1/2 less than 30 s) and rapidly reversible (t1/2 less than 3 min) at 37 degrees C. Binding was a saturable process with 75 +/- 12 fmol (-) [3H] alprenolol bound/mg protein (mean +/- SEM) at saturation, corresponding to about 2,000 sites/cell. Half-maximal saturation occurred at 10 nM (-) [3H] alprenolol, which provides an estimate of the dissociation constant of (-) [3H] alprenolol for the beta-adrenergic receptor. The beta-adrenergic antagonist, (-) propranolol, potently competed for the binding sites, causing half-maximal inhibition of binding at 9 nM. beta-Adrenergic agonists also competed for the binding sites. The order of potency was (-) isoproterenol greater than (-) epinephrine greater than (-)-norepinephrine which agreed with the order of potency of these agents in stimulating leukocyte adenylate cyclase. Dissociation constants computed from binding experiments were virtually identical to those obtained from adenylate cyclase activation studies. Marked stereospecificity was observed for both binding and activation of adenylate cyclase. (-)Stereoisomers of beta-adrenergic agonists and antagonists were 9- to 300-fold more potent than their corresponding (+) stereoisomers. Structurally related compounds devoid of beta-adrenergic activity such as dopamine, dihydroxymandelic acid, normetanephrine, pyrocatechol, and phentolamine did not effectively compete for the binding sites. (-) [3H] alprenolol binding to human mononuclear leukocyte preparations was almost entirely accounted for by binding to small lymphocytes, the predominant cell type in the preparations. No binding was detectable to human erythrocytes. These results demonstrate the feasibility of using direct binding methods to study beta-adrenergic receptors in a human tissue. They also provide an experimental approach to the study of states of altered sensitivity to catecholamines at the receptor level in man.


Localization of the membrane defect in transepithelial transport of taurine by parallel studies in vivo and in vitro in hypertaurinuric mice. We investigated the mechanism of taurinuria in three inbred strains of mice: A/J, a normal taurine excretor (taut+); and two hypertaurinuric (taut-) strains, C57BL/6J and PRO/Re. Plasma taurine is comparable in the three strains (approximately 0.5 mM), but taurinuria is 10-fold greater in taut- animals. Fractional reabsorption of taurine is 0.967 +/- 0.013 (mean +/- SD) in A/J); and 0.839 +/- 0.08 and 0.787 +/- 0.05 in C57BL/6J and PRO/Re, respectively. Taurine concentration in renal cortex intracellular fluid (free of urine contamination) is similar in the three strains. Taurine reabsorption is inhibited by beta-alanine, in taut+ and taut- strains. These in vivo findings reveal residual taurine transport activity in the taut- phenotype and no evidence for impaired efflux at basilar membranes as the cause of impaired taurine reabsorption. Cortex slices provide information about uptake of amino acids at the antiluminal membrane. Taurine behaves as an inert metabolite in mouse kidney cortex slices. Taurine uptake by slices is active and, at less than 1 mM, is greater than normal in taut- slices. Concentration-dependent uptake studies reveal more than one taurine carrier in taut+ and taut- strains. The apparent Km values for uptake below 1 mM are different in taut- and taut+ slices (approximately 0.2 mM and approximately 0.7 mM, respectively); the apparent Km values above 1 mM taurine are similar in taut+ and taut- slices. Efflux from slices in all strains in the same (0.0105-0.0113 mumol-min-1-g-1 wet wt), but taut- tissue retains about 10% more radioactivity over the period of efflux. beta-Alanine is actively metabolized in mouse kidney. Its uptake in the presence of blocked transamination, is greater; its intracellular oxidation is attenuated; and its exchange with intracellular taurine is diminished in taut- slices. These findings indicate impaired beta-amino acid permeation on a low-Km uptake system at the luminal membrane in the taut- phenotype. beta-Amino acids are not reclaimed efficiently either from the innermost luminal pool in cortex slices or from the ultrafiltrate in the tubule lumen in vivo. The former leads to high uptake ratios in vitro, the latter to high clearance rates in vivo. In vitro and in vivo data are thus concordant. This is the first time that a hereditary defect in amino acid transport has been assigned to a specific membrane surface in mammalian kidney.


Sequential activation of splenic nuclear RNA polymerases by erythropoietin. The spleen of the ex-hypoxic polycythemic mouse was employed to study the effect of erythropoietin on nuclear RNA polymerase activity. On the basis of ionic strength requirements and sensitivity to the fungal toxin alpha-amanitin, two major forms (I and II) of nuclear RNA polymerase were identified. Within 0.5 h after administration of erythropoietin, at a time when no morphologically identifiable erythroblasts were present in the spleen, there was an increase in the activity of polymerase II. By 2 h, polymerase II activity had declined to control levels. At 3 h, polymerase I activity began to increase, rising to a peak, 88% above control levels, by 12 h. During this period, early erythroblasts began to appear in the spleen. At 12 h, a second increase of similar magnitude occurred in polymerase II activity. Polymerase I activity fell to control levels by 18 h while polymerase II declined more slowly. These data indicate that stimulation of transcription is an early effect of erythropoietin. Multiple forms of RNA polymerase are involved and activation of these is sequential. Nuclear RNA polymerase activity is maximal during the period of early erythroblast proliferation and declines as these cells mature.


Clinical significance of serum properdin levels and properdin deposition in the dermal-epidermal junction in systemic lupus erythematosus. 61 biopsies of normal skin from the deltoid area and lesional skin from various sites from 48 patients with systemic lupus erythematosus (SLE) were studied for the presence of properdin, C3, C4, and immunoglobulins (IgG, IgM, and IgA) in the dermal-epidermal junction (DEJ) using direct and indirect immunofluorescence. Properdin was present in 50% of normal and 40% of lesional skins. Properdin was present without C4 in only 2 of 38 nonlesional skin biopsies and in only 2 of 20 lesions. There was no significant difference in incidence of deposition of any of the six proteins studied between nonlesional and lesional skin. The frequency of deposition of each of the proteins correlated with clinical disease activity. The presence of proteins in the DEJ did not correlate with the presence of active renal disease at the time of biopsy nor with previously documented active nephritis. In addition, no other single clinical manifestation correlated with the presence of DEJ deposition of any protein studied. IgA was not demonstrated in the DEJ of nonlesional skin of 16 patients in remission and was present in 7 of 23 patients with active disease (P less than 0.05). Deposition of properdin in lesional skin correlated with the presence of extracutaneous disease activity (P less than 0.05). Analysis of serologic studies on serum obtained at the time of biopsy revealed a statistically significant correlation between C4 and C3 (r = 0.67). This correlation was stronger than that between properdin and C4 (r = 0.37). Titer of antinuclear antibody and percent of DNA binding correlated better with C4 levels than with properdin levels. Serum properdin levels were significantly lower in patients with active disease than in those in remission (P less than 0.05). Serum properdin levels were significantly lower in patients with properdin deposits in lesional skin than in those without properdin deposits. The data suggest that both alternative and classical pathways are activated in patients with clinically active SLE.


The role and fate of rabbit and human transcobalamin II in the plasma transport of vitamin B12 in the rabbit. Previous studies have shown that plasma transcobalamin II (TCII) facilitates the cellular uptake of [57Co] vitamin B12 (B12) by a variety of tissues, but the lack of an intrinsic label on the protein moiety of the TCII-B12 complex has made it impossible to determine the role and fate of TCII during this process. We have labeled homogensous rabbit and human TCII with 125I-labeled N-succinimidyl-3-(4-hydroxyphenyl) propionate and have performed in vivo experiments in rabbits. When 125I-labeled rabbit TCII-[57Co] B12 and 131I-labeled bovine albumin were simultaneously injected intravenously, we observed that 125Iand 57Co were cleared from plasma at a faster rate (t1/2 = 1 1/2 h) than 131I and that 125I and 57Co were present in excess of 131I in the kidney, liver, spleen, heart, lung, and small intestine 1/2 h after injection. Later, 57Co remained in excess of 131I, but the ratio of 125I to 131I decreased progressively in all of these plasma and were rapidly excreted in the urine. After 1 h following injection, 57Co was present in excess of 125I in the plasma...


Blockade and stimulation of renal, adrenal, and vascular angiotensin II receptors with 1-Sar, 8-Ala angiotensin II in normal man. We have assessed the capacity of an analogue of angiotensin II (A II), 1-Sar, 8-Ala A II (P113) in normal man to stimulate and block responses to A II in four systems: blood pressure was monitored directly from an arterial catheter, and renal blood flow was measured with 133Xe and arterial renin and aldosterone concentrations by radioimmunoassay. The 31 normal subjects were in balance on a daily intake of 200 meg sodium and 100 meq potassium to suppress endogenous renin. P113 administered intravenously induced a dose-related renal blood flow reduction, with a threshold dose of 0.1 mug/kg/min. This dose also induced a small but significant increase in arterial blood pressure and plasma aldosterone as well as a reduction in plasma renin activity. In contrast to its effect on the renal vasculature, no tendency to a progressive response in the latter three parameters was noted as the P113 dose was increased 30-fold, to 3.0 mug/kg/min. P113 also reduced the clearance of para-aminohippurate, creatinine, sodium, and potassium, a pattern similar to that induced by A II. P113 at 0.1 mug/kg/min reduced significantly the blood pressure and renal vascular and aldosterone responses to graded doses of A II. Higher P113 doses totally obliterated all three responses to A II infused at 10 ng/kg/min, a dose that provides arterial A II concentrations in the range found in angiotensin-mediated hypertension. When A II was infused first, to induce a pressor, renal vascular, and aldosterone response, P113 induced a dose-related reversal of the response in each system. In conclusion, P113 is a partial agonist in normal man, inducing an angiotensin-like response in settings in which endogenous A II is not playing a tonic role, and displaying dominant antagonist activity in settings in which A II is active. Moreover, the studies suggest that the receptors mediating the responses to A II are different in the renal vasculature and other systemic vascular beds. The adrenal receptor must also differ. This agent should be useful in dissecting the role of A II in diseases characterized by hypertension or abnormalities of renal and adrenal function.


A renotropic system in rats.

A renotropic system in rats. While plasma or sera obtained from rats 20 h after removal of one kidney (uninephrectomy) stimulated [3H] thyrmidine incorporation into the DNA of kidney tissue incubating in vitro, azotemic plasma or sera obtained from rats 20 h after removal of both kidneys had no apparent effect. Dialysis of this azotemic sera resulted in its ability to stimulate isotope incorporation into renal DNA to the same degree as sera from uninephrectomized rats. This stimulatory factor (renotropin) was found to rise significantly within the first 26 h after uninephrectomy. Renotropin worked only on renal tissue, and we found that a factor  could be extracted in large amounts from the remaining kidney 20 h after uninephrectomy that would stimulate renal DNA synthesis in the presence of sera. Based on these findings and others, we postulate that after uninephrectomy there is an elevation in circulating renotropin as well as a tissue factor in the remaining kidney. Both factors together probably produce an excitor which enhances [3H] thymidine into DNA. The latter is tightly bound to renal tissue, and its production and/or activity is modified by circulating inhibitors that are especially prominent in azotemia.


The metabolism of supplementary vitamin C during the common cold. Ascorbic acid concentrations have been measured in leukocytes and plasma following oral administration of 2000 mg vitamin C in the same subjects while they had cold symptoms and after recovery from their colds. Plasma and leukocyte concentrations rose significantly in females, but only plasma concentrations rose in males, after the loading dose during colds. In the postcold tests, only plasma concentrations rose in both sexes. There was a significant difference in plasma leukocyte regression coefficients between the cold and postcold tests in females. Ascorbic acid passes into the plasma for metabolic purposes, and its storage is less in the leukocytes, during colds. Males had worse colds than females because their catarrhal symptoms were more severe. Higher tissue concentrations of ascorbic acid tended to be associated with low total, toxic, and catarrhal symptom values. A rise in tissue ascorbic acid was associated with less severe catarrhal symptoms in females. Ascorbic acid concentrations in the plasma and tongue were significantly higher after the subjects had recovered from their cold symptoms. Increasing the loading dose of vitamin C from 500 to 2000 mg more than doubled the leukocyte concentration of ascorbic acid in females. The higher dose enabled uptake of the vitamin into the leukocytes to take place over a 4-hour period. It did not give rise to increased uptake into male leukocytes. Administration of supplementary vitamin C elevated plasma ascorbic acid. The ascorbic acid then passed into the tissues depleted of vitamin C during the cold syndrome. A single supplementary dose of 2000 mg vitamin C can replete leukocyte ascorbic acid during a 4-hour period in females, but a larger dose may be necessary in males.


Effect of clonixin and aspirin on platelet aggregation in human volunteers. The effect of clonixin and aspirin on platelet function was assessed in healthy volunteers. Both drugs inhibited secondary platelet aggregation and prolonged bleeding time, but the effect of clonixin was significantly less than that of aspirin. Hemorrhagic complications are less likely after clonixin than after aspirin.


Influence of formulation on bioavailability of theophylline. The bioavailability of theophylline from formulations containing this drug alone and also in combination with phenobarbital and ephedrine has been studied in normal volunteers and asthmatic subjects. Neither of the latter two drugs had any significant influence on the bioavailability of theophylline. A complexation phenomenon, previously reported to inhibit dissolution of theophylline and phenobarbital in vitro, does not affect the bioavailability of theophylline from solid formulations. No differences were observed in the pharmacokinetics of theophylline between normal and asthmatic subjects.


Development of myelination in optic tract of the cat. The postnatal development of myelin in the optic nerve and tract of normal and dark reared cats has been studied quantitatively with light and electron microscopy. In the newborn cat few myelinated fibers (3% of the population) are seen in the optic tract. Until the end of the second postnatal week, the total number of myelinated axons in the tract remains low (23%). At this time, however, there is an explosive increase in the rate of myelination and by the end of the fourth postnatal week 80% of the optic tract axons have acquired a myelin sheath. Thereafter, the number of myelinated axons increases gradually, reaching adult levels (100%) at 12 weeks. During the initial period of myelination, the average axon diameter is 0.6 mu for unmyelinated fibers and a.2 mu for myelinated fibers. Both of these means remain substantially unchanged until myelination is completed, suggesting that initial myelination of an axon is not a continuous process but rather proceeds in a step-wise manner. Dark rearing appears to have no effect on the initiation of myelination.


Early formation of synapses in the molecular layer of the fetal rat cerebellum. The initial phase of synaptogenesis in the molecular layer of the albino rat cerebellum has been studied quantitatively and qualitatively with the electron microscope. Using both aldehyde-osmium (AL-OS) and ethanolic phosphotungstic acid (E-PTA) preparations, synapses were observed as early as embryonic day 19 throughout the entire rostro-caudal extent of the vermal molecular layer. The undeveloped nature of the elements participating in the formation of these synapses precluded their identification, for at this stage of cerebellar development, the Purkinje cells have not developed characteristic dendrites and both the massive production of cells within the external granule layer and the migration of neuroblasts through the molecular layer are in their initial phases. Evidence of synapse formation at this early stage of molecular layer development indicates that the rudiments of neuronal systems are present before the bulk of the molecular layer has come into existence and before most of the constituent elements of the cerebellum have differentiated. From embryonic day 19 to embryonic day 22, the number of E-PTA synaptic contacts was always less than the number of AL-OS synapses. This decrepancy and the variations observed in the morphology of the presynaptic components of E-PTA stained synaptic contacts, indicate that E-PTA morphology, and particularly the development of presynaptic dense projections, represents a late stage  of synapse formation.


Projections of the optic tectum in two teleost species. The efferent pathways of the optic tectum have been investigated in the percomorph Eugerres and the berycomorph Holocentrus. A portion of the dorsal-dorsolateral region of the optic tectum was unilaterally resected by suction. The animals were perfused 6-30 days thereafter, and the brains were processed according to a modification (Method 7 in Ebbesson, '70) of the Fink-Heimer ('67) technique for the selective silver impregnation of degenerating axons and terminals. Three groups of fibers emerge from the lesioned region (a) a medial group, which runs towards the midline and terminates in the ipsilateral torus longitudinalis and the contralateral tectum; (b) an ascending group, which enters the dorsocaudal region of the diencephalon and terminates in pretectal cell groups, in the dorsomedial optic thalamic nucleus, and in the nucleus rotundus or prethalamicus; and (c) a descending group, which funnels down into the midbrain tegmentum. Here abundant terminals are given to dorsolateral cell groups and to the nucleus isthmi. A recurrent fascicle leaves the mainstream and ascends to terminate in scattered diencephalic cell groups, in the nucleus geniculatus posterior pars ventralis, and in the nucleus rotundus or prethalamicus. The bulk of descending fibers then forms an ipsilateral bundle, which gives terminals to the lateral reticular formation of mesencephalon and rhombencephalon, and a contralateral (i.e., the predorsal) bundle, which terminates in the medial reticular formation of mesencephalon and rhombencephalon.


Telencephalic projections in two teleost species. The efferent pathways of the telencephalon were investigated in the percomorph Eugerres and the berycomorph Holocentrus. One telencephalic hemisphere was resected by suction and the animals were perfused 7-35 days thereafter. The brains were processed according to a modification (Method 7 in Ebbesson, '70) of the Fink-Heimer ('67) technique for selective silver impregnation of degenerating axons and terminals. Some fibers emerging from the lesioned telencephalic hemisphere terminate upon the contralateral hemisphere. The large bulk of efferent fibers, however, descends into the ipsilateral diencephalon and gives off the so-called strio-tectal bundle (STB) as well as the medial forebrain bundle (MFB). The remaining contingent eventually splits into the so-called strio-lobar bundle (SLB) and the lateral forebrain (LFB), but, previous to splitting, it contributes most of the telencephalic projection to the optic tectum in Eugerres, and gives abundant terminals to the ipsilateral nucleus rotundus or prethalamicus in both Eugerres and Holocentrus. The STB conveys all telencephalo-tectal fibers in Holocentrus, and some of them in Eugerres. Telencephalic efferents teminate ipsilaterally in the middle level of the tectum's stratum griseum centrale; in Holocentrus there is also a small projection to the stratum opticum. The MFB terminates at the caudal hypothalamus and gives terminals all along its course. The LFB also gives terminals all along its course and terminates upon a nucleus located between the midline and the corpus glomerulosum. The SLB spreads out and terminates in the inferior lobe of the hypothalamus.


Scotopic and photopic vision in the California ground squirrel: physiological and anatomical evidence. The California ground squirrel is a highly diurnal species previously thought to have an all-cone retina. This issue was re-examined in physiological and anatomical experiments. The electroretinogram (ERG) was used to measure the spectral sensitivity of the eye under different conditions of adaptation. The occurrence of a Purkinje shift could be demonstrated, although there was some indication that not all members of this species show such a shift. Spectral sensitivity of the dark-adapted eye of this squirrel is close to that predicted by a typical mammalian rhodopsin. Light adaptation produces a shift in spectral sensitivity to a peak location of about 525 nm. It was shown that two mechanisms having different spectral sensitivities contribute to the photopically recorded ERG. The degree to which these two mechanisms contribute to the ERG was found to be strikingly different from the degree to which the two contribute to visual behavior. Our anatomical results indicate that the retina of the California ground squirrel has two structurally distinct photoreceptors which, on the basis of various criteria, can be classified as cone and rod-like. The rod-like receptors comprise about 6-7% of the total. The two photoreceptor types differ in placement of their inner segments, size of their outer segments, outer segment ultrastructure, and terminal structure and organization.


Opossum somatic sensory cortex: a microelectrode mapping study. Organization of opossum somatic sensory cortex has been investigated utilizing closely spaced microelectrode penetrations (0.25-0.5 mm apart) and delicate mechanical stimulation of body surfaces including the facial vibrissae. Results may be summarized as follows: (1) the general organization of somatic sensory cortex, as originally defined by Lende ('63a) has been confirmed; (2) a double representation of the contralateral mystacial vibrissae and rhinarium, implicit in Lende's original data, was revealed in detail, the two representations being orderly, adjacent, mirror-images of each other; (3) units at a given cortical locus responded to deflection of between one and five mystacial vibrissae, about half responding to movement of a single vibrissa only; (4) about 40% of mystacial vibrissa units showed a directional specificity to the extent that they responded to deflections in only one or two cardinal directions; (5) units located in the medial vibrissa area showed a greater directional specificity than did units located in the lateral vibrissa area; (6) the surface area of rhinarial receptive fields was about ten times the area of first-order rhinarial unit receptive fields (B. Pubols et al., '73); (7) representation of the contralateral forelimb, especially the ventral surface of the forepaw, is extensive, orderly, and precise; (8) representation of the contralateral hindlimb, foot, and tail is minimal, and is confined to the midline convexity; (9) the presence of a small region of bilateral representation, lateral to the regions of contralateral representation, was confirmed. It is suggested that the region of contralateral postcranial representation plus the medial rhinarium and mystacial vibrissa areas are the homologue of SmI in placental mammals, and the region of bilateral representation is homologous to SmII of placental mammals, but that the lateral vibrissa and rhinarium areas are a specialization of somatic sensory cortex unique to the Virginia opossum.


Triamcinolone acetonide aerosols for asthma. I. Effective replacement of systemic corticosteroid therapy. Triamcinolone acetonide, a water-insoluble corticosteroid preparation in a Freon-propelled metered-dose aerosol, was administered via a specially designed nebulizer to 22 adult patients with severe, chronic asthma. All patients had required oral corticosteroid therapy, daily, for several years, in order to control their incapacitating obstructive airways disease. During 4 wk of treatment with triamcinolone acetonide, taken in 4 daily doses totaling 8 to 28 inhalations (400-1,400 mcg) per day: (1) asthma remained under satisfactory control; (2) oral corticosteroids were stopped in 19 patients and reduced in 2, and 1 patient withdrew from the study; (3) adrenal cortical function returned to normal or near normal levels; (4) spirometric and plethysmographic measurements improved significantly; (5) daily self-measurements of peak expiratory flow showed a marked improvement in median weekly levels, as well as a reduction in daily variability. No immediate undesirable side effects were noted. These observations indicated that more extensive applications of topical corticosteroid therapy are justified as an effective, convenient, and relatively safe method for the preventive management of severe, persistent asthma.


Comparison of respiratory and cardiovascular effects of isoproterenol, propranolol, and practolol in asthmatic and normal subjects. Pulmonary functions, by means of FEV1, FVC, and airway resistance, and cardiovascular responses, i.e., ECG, blood pressure, and pulse contour, were measured in 10 control and 15 symptom-free asthmatic subjects during and after the infusions of isoproterenol, with or without previous administration of propranolol or practolol. Bronchial sensitivity to methacholine and response to isoproterenol after methacholine were also measured. Bronchial sensitivity to methacholine challenge was markedly enhanced by propranolol and the effect of isoproterenol infusion on the pulmonary function was also significantly diminished, whereas practolol did not reveal any effect on bronchial sensitivity to methacholine or isoproterenol. The above changes were much more profound in symptom-free asthmatic subjects than in control subjects. The results suggest that practolol does not significantly affect the beta-2 bronchial receptors, is safe to use in asthmatic subjects, and is consistent with an abnormality of these receptors in bronchial asthma.


The relationship between HL-A antigens and lymphocyte response in ragweed allergy. The association between clinical ragweed hypersensitivity and HL-A haplotype inheritance has been confirmed. The lymphocyte response to phytohemagglutinin was greater in symptomatic individuals than in their asymptomatic relatives.


Studies on the deterioration of antigen E in extracts of Ambrosia elatior. Five per cent (w/v) extracts of defatted Ambrosia elatior pollen were prepared in bicarbonate-buffered saline (Coca's extract) and Coca's extract with 50% glycerol (Gly-Coca's extract). Sterile extracts were stored in untreated glassware and glassware coated with silicone. Stock extracts were diluted in phosphate-buffered saline (PBS) and stored at 5 +/- 3 degrees C. Extracts 1:20, 1:40, and 1:400 were assayed at approximately 6-wk intervals for antigen E. Quantitative determinations were made by modified double diffusion in agarose gels using known standards of antigen E for comparison. Results from the first year of this 3-yr study show no significant reduction in 1:20 Gly-Coca's extracts and a highly significant reduction of antigen E in 1:20 Coca's extracts. Further analysis of variance indicates significant reduction in all 1:40 and 1:400 extracts. Differences in the initial extracting solvents did not significantly affect these more dilute extracts. No significant differences were observed in the untreated and silicone-coated glassware. In terms of per cent antigen E reduction, 1:20 Gly-Coca's showed negligible reduction and Coco's at 1:20 declined nearly 50% in antigen E content. Extracts stored at 1:40 retained only 40% of the original antigen E content, while diluted extracts stored at 1:400 retained only 35% of the initial antigen E after 1 yr of storage at 5 +/- 3 degrees C.


Nutritional status of Head Start and nursery school children. I. Food intake and anthropometric measurements. Food intakes and anthropometric measurements of Head Start and nursery school children were compared in the fall and spring. Meals consumed at Head Start improved low calcium and ascorbic acid intakes. Iron intakes were low among both groups of children during both seasons. All children had high protein intakes in the fall and spring. Nursery school children consumed high levels of calcium and ascorbic acid and received more vitamin and mineral supplements. Anthropometric measurements indicated adequate and similar growth for all children during each season. Dietary intakes of mothers were less satisfactory than those of their children. Head Start mothers had the poorest diets, with frequent low intakes of energy, calcium, iron, vitamin A, ascorbic acid, riboflavin, and thiamin. Iron was the nutrient needed most by both groups of mothers. Nursery school mothers had especially high intakes of ascorbic acid, and consumed more vitamin and mineral supplements than Head Start mothers.


Nutritional status of Head Start and nursery school children. II. Biochemical measurements. At the beginning of a Head Start and nursery school program in the fall, nursery school children had significantly higher hemoglobin and hematocrit levels and mean corpuscular volume than Head Start children. The latter had a higher mean corpuscular hemoglobin concentration and alpha 1 globulin level. In the spring, at the close of both preschool programs, nursery school children still had higher hemoglobin and hematocrit levels and mean corpuscular volume, and also a higher mean corpuscular hemoglobin and total iron binding capacity. Head Start children exhibiting signs of anemia in the fall were improving in iron status in the spring. Normal blood protein levels for both groups of children indicated adequate protein status. This condition was confirmed by urinary analysis of urea, creatinine, and inorganic sulfate sulfur.


Evaluation of a ghetto school breakfast program. A program was initiated to provide a free breakfast at a ghetto elementary school. Community support was achieved at the expense of randomization of study subjects. While no significantly greater increase in school attendance or performance was detected in comparison with a control school, none of the students was malnourished and diets of students in the control school were as adequate as those in the breakfast school. Thus, it cannot be concluded that school breakfast programs would not benefit malnourished children or teen-agers who most often go without breakfast.


Effect of a low-carbohydrate diet and alcohol on perceptual motor skill. Eleven adults of both sexes ate a low-carbohydrate diet consisting mainly of meat, fish, and eggs for two days. After a low-carbohydrate breakfast on the third day, they operated a Stressalyzer to test their basic perceptual motor skill. Each subject drank 1.5 oz. rye whiskey at hourly intervals from 9:30 A.M. to 12:30 P.M. Blood glucose decreased to below 50 mg. per 100 ml. in three of the eleven subjects; two of the three had decreased perceptual motor skill.


Validity of the 24-hr. recall. Analysis of data obtained from elderly subjects. Tests of the validity of the 24-hr. dietary recall were done by comparing actual with recalled intakes for eight nutrients and the MAR (mean adequacy ratio) for a sample of seventy-six subjects age sixty years or older. Validity was tested by using paired-t tests and regression analysis. In the paired-t test, no significant difference was found between the mean recalled and the mean actual intake of nutrients, with the exception of calories. Using regression analysis, results indicated that for three of the eight nutrients considered (calories, protein, and vitamin A), small intakes tend to be over-reported and large intakes under-reported (p less than .05). Thus, for these three nutrients, the recall seems to be statistically conservative for group comparisons; it would seldom, if ever, indicate a difference in intake where no difference exists. But, it could yield a false negative, i.e., an indication of no significant difference, when, in fact, a difference does exist. Clearly, more research is needed, both to replicate this study and to develop techniques with greater internal validity for comparing the dietary intakes of groups.


Comments on dietary intake of drug-dependent persons. Interviews by means of a questionnaire were used to study the food habits and estimate the nutritional value of food eaten by forty clients in a methadone maintenance program and nineteen clients in a drug detoxification program in a drug treatment center, Marin Open House, San Rafael, California. Although responses to the questionnaires were highly varied, many subjects reported that drugs did affect appetite or cause craving for certain foods. Patients in the detoxification program tended to eat meals either once or twice a day, whereas one-third of those in the methadone program ate three or more meals daily. Over 90 per cent of the clients reported eating snacks. Foods consumed in inadequate amounts included fruits and vegetables high in either vitamin A or ascorbic acid, liver and other meats, and whole grain and enriched cereals or bread. Food consumed as reported on a 24-hr. recall was evaluated by a computer program, and the tabulation was compared with the Recommended Dietary Allowances. Diets were considered to be inadequate if nutrient intakes were below two-thirds of the allowances. In general, the clients in the detoxification program consumed less adequate amounts of the nutrients studied than the methadone group. The females in both groups consumed less adequate diets than the males. About two-fifths of the men of both groups and about three-fourths of the women had diets inadequate in three or more nutrients by the standards chosen. In spite of the limitations of this study, the results do point to the existence and type of nutritional problems likely to be encountered in drug-dependent persons and in those undergoing treatment. The need for nutrition counseling is evident.


C3 nephritic factor (C3NeF): stabilization of fluid phase and cell-bound alternative pathway convertase. C3 nephritic factor (C3NeF) defined by the capacity of nephritic serum and its fractions to initiate loss of the B antigen of C3 in normal serum was purified from the serum of three different donors and shown to function by stabilization of membrane-bound and fluid phase alternative pathway C3 convertase. C3NeF converts cell-bound C3B sites in a dose-related manner to CEB(NeF) sites, which exhibit an approximate 10-fold increase in half-life. The linear relationship between the C3NeF input and the residual hemolytic sites on EAC43B present after incubation for 20 min at 30 degrees C, during which labile C3B sites have decayed, indicates that the number of residual C3B sites is directly related to the dose of C3NeF. The capacity of C3NeF to stabilize the C3B convertase in a temperature- and dose-dependent manner, which is independent of binding or consumption of C3NeF, in a fluid phase reaction mixture of 125I-B, 131I-C3 and D permits isolation of a 10S complex containing radiolabeled C3 and B and exhibiting C3, convertase activity on an exogenous C3 source. Thus, the stabilizing effect of C3NeF is not limited to membrane-bound C3B but is also sufficient to permit recovery of a fluid phase C3 convertase formed during the interaction of C3, B, and D.


Effect of cold exposure on the metabolism of immunoglobulins in rabbits. The effect of low environmental temperature on the metabolism of IgG and IgM was examined in unimmunized rabbits. The half-lives of both IgG and IgM were less in animals kept at 4 degrees C for 6 weeks than in animals kept at 22 degrees C. Serum concentration of IgM and GG were unaltered by cold exposure but intravascular pool sizes tended to increase as a consequence of an expanded serum volume. Fractional turnover rates of both IgM and IgG were greater in cold-exposed animals. At both 4 degrees C and 22 degrees C, the fractional catabolic rate of IgM was independent of its serum concentration whereas that of IgG was correlated directly with its serum concentration. Absolute turnover of both IgM and IgG was accelerated by cold exposure. It is suggested that increased synthesis of immunoglobulin could account for the higher levels of antibody reportedly found in cold-exposed rabbits.


Cell-mediated immune response to mumps virus infection in man. The antibody and cell-mediated immune response to mumps virus infection was studied in groups of subjects after natrually acquired mumps virus infection, after parenteral immunization with live attenuated mumps vaccine, and in a population of mumps seronegative subjects. The technique of neutralization of tissue culture infectivity was utilized to study mumps specific antibody. The cell-mediated immunity (CMI) was detected by specific immune release (SIR) of radioactivity by purified lymphocytes after they were reacted with radioactive chromium (51Cr) labeled human conjunctival cell cultures chronically infected with mumps virus. No SIR activity was observed in lymphocytes obtained from cord blood and young individuals seronegative for antibody to mumps virus. Detectable SIR activity was observed in a few older seronegative subjects; however, immunization with mumps vaccine in such antibody negative subjects failed to result in the development of any antibody response in the serum. High SIR activity was observed in the lymphocytes of naturally infected and vaccinated subjects. Although all naturally infected or immunized subjects had varying levels of mumps specific antibody activity in the serum, no correlation existed between the levels of antibody and SIR activity. These observations suggest the development of mumps specific in vitro correlates of CMI after naturally acquired or vaccine-induced mumps virus infection.


Effect of VHa locus allotype-suppression on the expression of closely linked VHx, VHy, and Cmun genes in heterozygous rabbits. Injection of a1x-y-n81/a2x32y33n82 heterozygous rabbit at birth with anti-a2 allotype antiserum suppresses the production of the VHa2 gene product a2 Ig as well as the expression of the closely linked Cmun gene product n82 but not of the closely linked VHx and VHy gene products x32 and x33. This effect is related to the appearance of a2 and n82 on the same IgM molecules whereas a2, x32, and y33 appear on different Ig molecules.


Clq inhibition of the interaction of collagen with human platelets. Human Clq, isolated in pure state after affinity chromatography on IgG-Sepharose, inhibited collagen-induced aggregation and release of 14C-Serotonin from prelabeled human platelets. Platelet aggregation induced by ADP or thrombin was not inhibited by Clq. Also, the adherence of platelets to glass surfaces was significantly diminished by Clq. In contrast, aggregated Clq mimicked the effect of collagen in causing platelet aggregation and release of serotonin. It appears that monomeric Clq, which has structural similarities to collagen competes with collagen for specific sites on the platelet surface.


Number of macrophages and distribution of mitotic activity in regressing and progressing Moloney sarcomas. The concentration of macrophages per gram tumor was as much as five times greater in regressing compared to progressing Moloney sarcomas. Infiltrating monocular cells, including macrophages, were closely associated with a cessation of mitotic activity in tumors.


Loss of carrier-determined tolerance in vitro with loss of receptor blockade. Experiments were done to determine whether carrier-determined tolerance is reversible and whether the loss of tolerance is accompanied by the loss of receptor blockade. Spleen cells from mice made tolerant with DNP-isologous IgG remained tolerant when transferred to irradiated syngeneic mice. If these same tolerant spleen cells were incubated for 24 hr or more before transfer the tolerance was lost. Autoradiology was done on the tolerant cells with either 125I anti-DNP or 125DNP-KLH, before and after incubation in vitro. When the cells were tolerant the number of DNP ABC was decreased whereas cells having DNP on their surface were increased. When the cells lost tolerance after in vitro incubation, the hapten-bearing cells were no longer present although the number of cells free DNP receptors increased to normal. These data suggest that in carrier determined tolerance the reactivation of tolerant lymphocytes may involve reversible receptor blockade.


Detection of immune complexes in unheated sera by modified 125I-Clq binding test. Effect of heating on the binding of Clq by immune complexes and application of the test to systemic lupus erythematosus. The 125I-Clq binding test was modified in order to allow for the detection of immune complexes in native unheated human serum. Indeed, heat-inactivation (56 degrees, 30 min) was found to reduce the Clq-binding activity of immune complexes mixed with native serum. This effect was not observed when EDTA was added to the native serum before mixing the immune complexes. The modified 125I-Clq binding test was performed in two steps: first, the tested native serum sample was incubated for 30 min at 37 degrees C with 0.13 M EDTA in order to prevent the integration of 125I-Clq into the intrinsic Clqrs complex, second, 125I-Clq and polyethylene glycol (final concentration 2.5%) were added to this mixture, and further incubated for 1 hr at 4 degrees C. Under these conditions, free Clq remained soluble whereas Clq bound to macromolecular complexes was precipitated. The competitive effect of intrinsic Clq and the interference of other substances such as DNA or bacterial LPS were very limited. The modified Clq binding test was applied to the clinical investigation of 44 patients with systemic lupus erythematosus; and increased Clq binding activity (Clq-BA) was observed in 91% of the samples. The level of Clq-BA was found to be significantly correlated to the DNA-binding capacity and to the decrease of the level of some complement components.


Antibody-mediated cell cytotoxicity in a defined system: regulation by antigen, antibody, and complement. The use of a serum-free environment and target cells carrying defined amounts of radiolabeled antigen allowed a quantitative study of the role of antigen, antibody, and complement on antibody-mediated cell cytotoxicity (AbMC). For lysis to occure, a minimum number of antigen molecules must be present on the target cell. 51Cr release from target cells with lower antigen density requires larger concentration of effector cells and antibodies. Target cell-bound complement, itself unable to mediate cytotoxicity, reduces the number of IgG molecules required for lysis. The antibody and complement, however, have to be bound to the same target cell. Bystander complement-coated erythrocytes, present in the same reaction mixture with IgG-coated targets, are not lysed. Blocking of AbMC is effected only by antigen, either soluble or in immune complexes prepared in antigen excess. Antigen competes at the level of the target cell. Blocking at the level of the effector cell, by use of immune complexes prepared at equivalence or in antibody excess, is difficult to achieve. The large number of cells with Fc receptors contained in mouse spleens may explain this finding. Arming of effector cells by passive binding of immune complexes is poorly effective as a means of obtaining lysis of the target cells. In all situations, the outcome of the reaction is determined by the presence of free antibody-combining sites, alone, or in immune complexes, that are able to combine with the target cell membrane antigen. The requirements for lysis are rather stringent.


The effects of external K+ and Na+ on the chemotaxis of rabbit peritoneal neutrophils. Extracellular K+ enhances the chemotactic responsiveness  and spontaneous movememt of rabbit peritoneal neutrophils but is not required for these functions. Other monovalent cations act the same; the rank order of their effectiveness is K+ = NH4 greater than Rb+ Cs+ greater than Li = Na. The K+ specific ionophore, valinomycin (10-7 M) inhances chemotaxis in the presence of K+ but not in its absence; another K+ specific ionophore, nigericin (10-7 M) inhibits chemotaxis in the absence of K+ but not in its presence. Ouabain (5 x 10-6 M) prevents the enhancing effects of K+ on chemotaxis. Removing the Na+ of the buffer and substituting it with K+, choline or glucose greatly enhances spontaneous motility but depresses chemotactic activity. One hypothesis suggested by the above results is that as a part of their action, chemotactic factors stimulate a net influx of K+ into the neutrophil; an alternative or additional hypothesis is that chemotactic factors stimulate a net efflux of Na+ from the neutrophil.


Factors affecting human percutaneous penetration of methotrexate and its analogues in vitro. Factors affecting the percutaneous penetration of methotrexate (MTX) and its analogues through excised human skin were studied. With application of 0.05% MTX solution to the epidermis, 0.07% of the applied dose was recovered in the epidermis and less than 0.005% penetrated completely through the skin. The percent of drug which penetrated remained constant from 0.05% to 2.0% MTX and was also unchanged by increasing incubation temperature from 28 degrees C to 37 degrees C. Cellophane tape stripping of stratum corneum following drug incubation removed essentially all the MTX that had been measured in whole epidermis. Binding of MTX was localized to the outermost layers of the stratum corneum. With removal of the stratum corneum prior to drug incubation, 25% of the applied MTX penetrated through the skin. Various vehicles were tested to determine their effect on MTX percutaneous absorption. Dimethylsulfoxide (80%), dimethylacetamide (25%), or retinoic acid (0.1%) had no effect on penetration. Maximum enhancement of penetration by 25-fold and 143-fold was obtained with retinoic acid (saturated solution in aqueous ethanol) and C-10-methylsulfoxide (2.5%), respectively. The penetration of lipid-soluble derivatives of MTX was also tested. The compound that penetrated best was the dimethyl ester of dichloro-MTX with a 3-fold increase in penetration over MTX.


Distribution of cholesterol-5alpha,6alpha-epoxide formation and its metabolism in mouse skin. The distribution of cholesterol-5alpha,6alpha-epoxide in ultraviolet light-irradiated mouse skin was determined. Highest levels of cholesterol-5alpha,6alpha-epoxide were found in the epidermis and distributed rather evenly throughout all subcellular fractions. Low but potentially significant levels of the compound diffused from the epidermis. Metabolic studies demonstrated that cholesterol-5alpha,6alpha-epoxide was converted to cholestane-3ss,5alpha,6ss-triol by an enzyme, cholesterol-5alpha,6alpha-epoxide hydrase, which was localized in the epidermal fractions sedimenting at 10,000 and 100,000 X g. Comparative studies indicated a much higher capacity for metabolism of cholesterol-5alpha,6alpha-expoide in liver than in skin. The effect of the epoxide and its metabolic product on the skin is discussed.


The induction of human multinucleated monocytes in culture. The formation of multinucleated giant cells in vitro has been studied using human monocyte cultures. Multinucleated monocytes appeared early in the course of the culture, indicating that they are not purely a phenomenon found only in aging cultures. In 4 of the 33 normal adults, more than 20% of multinucleated forms appeared in the cultures. Supernatants from early monocyte cultures of those subjects having a high incidence of multinucleated cells induced the formation of multinucleated cells in other cultures. This effect did not occur when supernatant from subjects with low counts of multinucleated cells were incubated with other monocyte cultures.


Phosphonoacetic acid treatment of shope fibroma and vaccinia virus skin infections in rabbits. The antiviral efficacy of phosphonoacetic acid (PAA) was studied in localized skin lesions of rabbits produced by the intradermal inoculation of vaccinia virus (VV) and of Shope fibroma virus (SFV). Systemic administration of PAA by intraperitoneal injections had no significant effect on the pustular lesions induced by VV or on the benign skin tumors caused by SFV. A complete suppression of the appearance of VV-induced pustular lesions was achieved by 2% PAA ointment applied twice daily for 4 days, starting 24 hr after virus inoculation. A significant effect against SFV-induced tumors was obtained by PAA ointment applied beginning either 24 or 72 hr after virus inoculation. A complete suppression of SFV-induced tumors was observed when a dose of 10 mg PAA was injected intralesionally once daily for 5 days, beginning treatment 24 hr after virus inoculation. A significant reduction of the intensity of the tumors was seen following the same treatment schedule but with a delay of 72 hr after virus inoculation or by reducing the length of treatment to 3 days or with a dose of 1 mg injected intradermally daily for 5 days. After the healing of the lesions, PAA-treated rabbits were resistant to reinfections to the same extent as those in which spontaneous healing had occurred.


Inhibition of the in vivo effects of concanavalin-A on mammalian epidermis by alpha-methyl-D-glucopyranoside. Concanavalin-A (Con-A) injected intradermally into newborn rats produces inhibition of granular-cell formation, accumulation of spinous cells, glycogen deposition, and a decrease followed by an increase in the number of basal cells in DNA synthesis. These changes were maximal with a dose of 0.1 mg Con-A, although 0.005, 0.01, and 0.05 mg caused some epidermal changes. The Con-A effects were partially blocked when 0.1 ml of 0.3 or 0.1 M alpha-methyl-D-glucopyranoside (alphaMG) solution was injected 2 hr after 0.1 mg Con-A and completely inhibited by injection of 0.1 ml of 3.0 M alphaMG solution. The inhibitory effects were not seen after injection of 0.1 ml of 3.0 M N-acetyl-galactosamine saline solution, or 0.1 ml normal saline. Injection of alphaMG alone did not cause any changes in epidermal cells. These results indicate that specific sugar inhibits Con-A effects on mammalian epidermis in vivo.


Skin in dermatosparaxis. Dermal microarchitecture and biomechanical properties. Dermatosparaxis is a heritable disorder of the connective tissue observed in the calf and is related to the presence in the dermis of a form of procollagen, rather than collagen, forming the fibrous support of the skin. This alteration results from the lack of activity of procollagen peptidase, an enzyme required for the final processing of procollagen in collagen. Skin in dermatosparaxis displays abnormal mechanical properties. Procollagen forms poorly structured fibers loosely packed in sheets and dispersed within an excess of glycosaminoglycans. This disturbed architectural organization is responsible for the hyperextensibility and poor elasticity of the dermatosparactic dermis. Skin fragility is related to the impaired mechanical properties of the procollagen polymers. Rupture occurs in the mid-dermis in a location where the connective tissue is devoid of adnexae and elastic fibers.


Monocytopoiesis in chronic eczematous diseases, psoriasis vulgaris, and mycosis fungoides. Monocytopoiesis and blood monocytes were examined in 8 patients with disseminated chronic eczematous diseases, 8 patients with disseminated psoriasis vulgaris, and 8 patients with mycosis fungoides in plaque or tumor stage. Monocytopoiesis was moderately stimulated in all these patients. The stimulation manifested itself by: (1) a rise in relative number of promonocytes in bone marrow in all patients with eczema, in 1 out of 8 patients with psoriasis, and in 7 out of 9 examinations in patients with mycosis fungoides; (2) a rise in [3H]thymidine labeling indices of medullar promonocytes (8/8 eczema, 7/7 psoriasis, 8/9 mycosis fungoides); and (3) a rise in the naphthol-AS-D-chloroacetate esterase activity of blood monocytes, indicating premature monocyte marrow egress (3/5 eczema, 7/8 psoriasis, 9/9 mycosis fungoides). In eczema and psoriasis the mean enhancement of monocytopoietic activity was similar but less pronounced than in mycosis fungoides. In the latter disease there was no correlation between measured parameters and visible skin lesions. The results were interpreted as indicative of increased monocyte consumption by pathologic, immunologic, and/or inflammatory processes.


Dinitrochlorobenzene responsivity in disseminated coccidioidomycosis: an inverse correlation with complement-fixing antibody titers. The immunologic status of 25 patients with disseminated coccidiodomycosis was evaluated by serum anticoccidioidin complement-fixing antibody levels, coccidiodin skin tests, and dinitrochlorobenzene (DNCB) sensitization. In the 10 patients who had disseminated disease and a complement-fixing titer of 1:32 or less, responses to DNCB were similar to those of 20 controls. In the 15 patients with disseminated disease and a complement-fixing titer of 1:64 or more, responses to DNCB were statistically significantly diminished compared to controls (p = 0.002). Since the complement-fixing titer is associated with extent of dissemination, these results signify a relationship between diminished DNCB responses and extensive dissemination. Of the several hypotheses which might explain this relationship, we find the most attractive is that of a nonspecific deficiency of cell-mediated immunity developing secondarily to extensive disseminated disease.


Mechanism of reappearance of contact sensitivity to DNCB in desensitized guinea pigs. The different immunologic activities of lymph node and spleen cells as compared to peritoneal exudate cells were studied in temporarily desensitized, DNCB-contact-sensitive guinea pigs. Lymph node (and spleen) lymphocytes: (a) transfer contact sensitivity to syngeneic recipients when harvested at the time of unresponsiveness; (b) are only temporarily inactivated in tolerant recipients by the desensitizing procedure; (c) enter the lymph node (or spleen) compartments in a higher proportion than peritoneal exudate cells; (d) are not susceptible to inactivation in vitro; and (e) are not permanently desensitized when released into the circulation by an intravenous injection of Bordetella pertussis vaccine. These results indicate that the resistance of primed lymph node cells to the desensitizing procedure may account for the reappearance of contact sensitivity in desensitized guinea pigs.


Recurrent bacterial infections in four siblings with neutropenia, eosinophilia, hyperimmunoglobulinemia A, and defective neutrophil chemotaxis. Four siblings with recurrent bacterial infections, neutrophil chemotactic defect, neutropenia, and eosinophilia were studied. During periods of infection the peripheral neutrophil count increased to normal, while the eosinophilia disappeared. In addition, these children had high levels of serum IgA and poor antibody responses to tetanus and polio vaccinations. A defect in cell-mediated immunity was demonstrated by an absent or weak reactivity to various skin test antigens and by abnormal lymph node histology. Thus these siblings had an unusual combination of defective inflammatory response and immunologic abnormalities.


Effects of cytosine arabinoside, adenine arabinoside, and 6-azauridine on rabies virus in vitro and in vivo. The antiviral agents cytosine arabinoside, adenine arabinoside, and 6-azauridine were shown to inhibit the replication of rabies virus in vitro but not the replication of Sindbis virus. These same drugs were not effective in reducing the mortality rate in mice challenged with street rabies virus.


Arthritis in mice due to infection with Mycoplasma pulmonis. II. Serological and histological features. Histological and serological data were obtained from a group of 160 mice given intravenous inoculations of 3.5 X 10(7) colony-forming units of Mycoplasma pulmonis and studied for 46 weeks. The basic pathology was that of an initial acute inflammatory response within the joint and periarticular tissues, slowly changing to a chronic inflammatory response seen predominantly in subsynovial areas with marked synovial proliferation. Occasional lymphoid follicles and plasma cells were seen in the subsynovial spaces. Titers of antibody to M. pulmonis were determined by both metabolic inhibition and indirect hemagglutination techniques. By the metabolic inhibition method, titers were highest between two and 22 weeks, and by indirect hemagglutination between four and 42 weeks. Antibodies to synovial antigens were present between nine and 26 weeks as determined by indirect hemagglunination. Serologic cross-reactions were found between M. pulmonis and normal mouse synovium in sera from infected mice and, by indirect hemagglutination, metabolic inhibition, and immunodiffusion techniques, in sera of rabbits or mice immunized with either M. pulmonis or mouse synovium.


Immunochemistry of otitis media with effusion. For investigation of the nature and origin of middle ear effusions, immunochemical studies were performed on more than 400 patients diagnosed as having otitis media with effusion. Although results of cellulose acetate and disc electrophoretic analyses and quantitation of IgG, IgA, and IgM suggested that proteins found in the effusions were derived for the most part from the serum, quantitative analysis of secretory IgA revealed the existence of appreciable amounts of secretory IgA in both serous and mucoid effusions. The antigenicity and subunit structure of the secretory IgA isolated from middle ear effusions were identical or very similar to those of secretory IgA obtained from other external secretions. Radioactive single radial diffusion analysis of IgE showed that the mean concentrations of IgE in effusions and sera were within normal ranges. Findings of this study suggest that the middle ear maintains the local immunologic defense system, that the middle ear effusion is at least partially an external secretion, and that IgE in middle ear effusions obtained from old patients, rather than being a local product, may be derived from the serum.


Attenuated influenza virus in normal adults: role of pulmonary function studies in vaccine trials. A live attenuated virus, influenza A/Hong Kong/68(H3N2)-ts-1[E], was administered to 10 normal volunteers. Infection resulted in minor upper respiratory tract symptoms in one volunteer. An isolate from one volunteer contained a presumed revertant wild-type virus, but no clinical significance could be ascribed to this infection. Since infection with natural influenza A virus adversely affects airway dynamics, serial tests of pulmonary function were performed, including measurements of forced expiratory flow rates, lung volumes, diffusing capacity, closing volume, and total pulmonary resistance at various frequencies by the oscillometric technique. All tests performed before and on days 3, 30, and 60 after administration of virus yielded normal results throughout the study; these results suggested that there was no inflammatory response to the candidate vaccine in either the large or the small airways. Determination of total pulmonary resistance by the oscillometric technique proved to be a simple and acceptable procedure and may be useful in large-scale epidemiologic studies.


Comparison of amphotericin B and amphotericin B methyl ester: efficacy in murine coccidioidomycosis and toxicity. The methyl ester of amphotericin B was compared with the parent compound, amphotericin B, in terms of therapeutic efficacy in experimental murine coccidioidomycosis and of toxicity. Infections were established by intraperitoneal or intratracheal inoculation with arthrospores. The mice were given either intraperitoneal or intravenous injections of drug for 30 days, according to several dosage schedules. At low doses, amphotericin B methyl ester was less effective therapeutically than was amphotericin B; however, at higher doses, amphotericin B was directly lethal and/or nephrotoxic, whereas the methyl ester was therapeutically effective and nontoxic. In contrast to amphotericin B, the methyl ester did not cause either azotemia or histopathologic changes in the kidneys.


Uptake of antibiotics by human erythrocytes. Human erythrocytes have been used as a model system for the study of uptake and release of antibiotics. Penicillin G, dicloxacillin, tetracycline, and minocycline were all taken up by the cells, but each showed a characteristic ratio of distribution between the extracellular and intracellular compartments. Comparison of the penicillin analogues indicated that dicloxacillin, the more lipid-soluble compound, reached higher intracellular concentrations than did penicillin G. Use of human plasma as the incubation medium markedly decreased antibiotic uptake and enhanced antibiotic egress from preloaded cells. These effects were related to the binding of drugs to serum proteins. In vivo studies in which penicillin G was injected intravenously by bolus and was then given by constant infusion showed that intraerythrocytic concentrations of drug after 2 hr approached or exceeded those in plasma. These results can be explained, in part, by the fact that the slower rate of efflux of penicillin G from the cells than of clearance from plasma serves to maintain the high initial levels of drug for a longer period.


Current status of passive immunity to diphtheria and tetanus in the newborn. The extent of transplacentally derived immunity to the toxins of Corynebacterium diphtheriae and Clostridium tetani was assessed by a hemagglutination technique using the cord sera of a group of infants recently born in Bronx, New York. Protective levels of antitoxin to C. diphtheriae were found in 64% of the infants. Tetanus antitoxin was present in sufficient quantity for protection in 38% of the cord sera. While this degree of immunity reepresents an improvement over that achieved 20 years ago and probably reflects the effects of immunization procedures, further attention to the maintenance of immunizations in women of childbearing age would improve these statistics.


Synergistic action of amphotericin B and rifampin against Candida species. Amphotericin B and rifampin act synergistically against certain yeasts in vitro. Whether this synergism is a general phenomenon or whether the effect has strict species and strain requirements was studied. Included in a survey of the genus Candida were eight human isolates of Candida albicans and one strain each of Candida krusei, Candida tropicalis, Candida pseudotropicalis, Candida parapsilosis, Candida guilliermodnii, and Candida stellatoidea. Cultures in both control and drug-containing liquid medium were incubated at 37 C with aeration. Effects of the drugs were determined from viability assays performed at zero-time and at 17 hr. Amphotericin C and rifampin were judged to be synergistic if any one of three different sets of criteria was met. Combined activity greater than the sums of individual drug effects was required in each set of criteria. Partially inhibitory or fungistatic levels of amphotericin B and noninhibitory concentrations of rifampin acted synergistically against all strains of Candida examined. Within the genus Candida, synergism of amphotericin B and rifampin appears to be a rather general phenomenon.


Radioimmunoassay of human C-reactive protein and levels in normal sera. C-reactive protein (CRP) has been considered an acute-phase protein which appears only during reactions of tissue injury or inflammation. We report here the quantitation of CRP levels in normal adults and neonates made possible by the development of a sensitive and precise radioimmunoassay for human CRP which enables the detection of 3 ng. per milliliter. Serum levels in 153 healthy blood donors ranged from 68 to 8,200 ng. per milliliter, with a median value of 580 ng. per milliliter (mean = 1,340 ng. per milliliter). CRP levels in 24 normal cord serum samples ranged from 10 to 370 ng. per milliliter, with a median value of 70 ng. per milliliter (mean = 109 ng. per milliliter). CRP levels in 246 individuals evaluated for autoimmune diseases ranged to 256,000 ng. per milliliter with a median value of 13,000 ng. per milliliter (mean = 38,000 ng. per milliliter). No individual lacking CRP was detected. Thus, CRP can be considered a component of normal serum which only increases dramatically in concentration during inflammation.


Quantitative evaluation of serum pancreatic isoamylases in cystic fibrosis. The amylase in sera of 20 patients with cystic fibrosis of the pancreas (CFP), 17 of whom had pancreatic insufficiency (PI), and of 34 normal control subjects was separated into its pancreatic and salivary components and quantitated. Electrophoretic separation was achieved on polyacrylamide disc gels with subsequent sectioning of the gel columns. The amylase content in international units per liter (IU/L.) was obtained for each section by the insoluble blue starch substrate method. The mean total serum amylase of the CFP patients with PI was significantly lower than the control group. A highly significant decrease was found when the pancreatic isoamylase was considered alone. The salivary isoamylase level remained essentially the same. Three patients with CFP but without PI demonstrated higher than normal pancreatic amylase values in the serum. These patients may be in an early stage of a progressive involvement of the pancreas in which there is an increased release of amylase. This method of electrophoretic separation and quantitation of the serum amylase is sensitive and efficient and allows the simultaneous processing of multiple specimens. The results obtained agree with previously published ratios of serum isoamylases. Our method should be valuable in the study of other diseases of the exocrine pancreas and salivary glands.


Satisfactory radioiodination of rabbit fibrinogen using chloramine-T. A method for radioiodination of fibrinogen using chloramine-T is described. Samples of greater than 90 per cent clottable counts were obtained. Electrophoretic mobility and immunodiffusion showed that the entire radioactivity was present in the fibrinogen band. In vivo studies on the turnover of this labeled product in rabbits showed a half-life of 52.8 to 61.7 hours in two batches of animals. The results compare very well with the reported results obtained from fibrinogen labeled with radioactive iodine by the iodine-monochloride method. The advantages of the new method are indicated.


Clot retraction: evaluation in dilute suspensions of platelet-rich plasma and gel-separated platelets. Suspensions of platelet-rich plasma (PRP) or gel-separated platelets (GSP) can be used to evaluate clot retraction subsequent to platelet aggregation and fibrin formation. PRP (200,000 per cubic millimeter) or GSP (200,000 or 100,000 per cubic millimeter) are diluted 1:10 (PRP) or 1:8 (GSP) in phosphate buffer, pH 7.4, and clotted with a high concentration (2.5 U. per milliliter) of thrombin. Human fibrinogen (25 mg. per cent) is added to GSP prior to dilution. Clot retraction is 91 to 100 per cent completed in 1 hour and is quantified by measurement of residual fluid volume. Test conditions are unfavorable for fibrinolysis. Very low concentrations of fibrin/fibrinogen degradation products D and E are detected in residual fluid, and no erythrocyte fall-out occurs. Furthermore, the extent of retraction in the dilute systems is related only to platelet numbers and platelet function. The dilute PRP and GSP methods allow evaluation of clot retraction in the presence of PGE1, the most potent inhibitor of platelet aggregation induced by conventional concentrations of collagen, ADP, epinephrine, and thrombin (0.1 to 0.5 U. per milliliter). High concentrations of PGE1 (to 6 x 10(-6) M) do not inhibit aggregation of GSP, fibrin formation, or platelet-fibrin interaction induced by 2.5 U. per milliliter of thrombin. In contrast, PGE1 concentrations as low as 1.5 to 3.0 x 10(-8) M inhibit clot retraction in both the dilute PRP and GSP systems. Thus, using dilute PRP or GSP the effects of platelet aggregation inhibitors on clot retraction can be determined independently of effects on platelet aggregation.


Influence of renin depletion on renal function after release of 24-hour ureteral obstruction. A marked increase in preglomerular resistance mediated through the renin-angiotensin system has been proposed as the mechanism for the sustained decrease in glomerular filtration rate seen following release of 24-hour ureteral obstruction. The importance of the renin-angiotensin system in mediating this response was evaluated by determining whole kidney and single nephron function following release of 24-hour ureteral obstruction in rats with normal renal renin content and in rats depleted of renal renin by desoxycorticosterone acetate acetate (DOCA) and saline treatment. The DOCA and saline treatment was effective in reducing renal renin content to less than 10% of the normal values. However, when compared to nonobstructed kidneys, both whole kidney filtration rate and single nephron filtration rate were similarly and significantly reduced in both groups following release of 24-hour ureteral obstruction. Single nephron stopflow techniques were used to determine the net hydrostatic force for filtration. The net hydrostatic force for filtration in control nonobstructed nephrons averaged 37.8 +/- 1.1 mm. Hg, but was significantly decreased to 22.5 +/- 2.2 mm. Hg in the normal renin obstructed kidney and to 18.8 +/- 1.0 mm. Hg in the renin-depleted obstructed kidney. It is concluded that the marked depression in glomerular filtration rate seen following release of 24-hour ureteral obstruction is due to increased afferent arteriole resistance and that the renin-angiotensin system is apparently not important in mediating the response.


Labile aggregation stimulating substance, free fatty acids, and platelet aggregation. Labile aggregation stimulating substance (LASS), an intermediate produced during platelet biosynthesis of PGE2 and PGF2alpha, acts as a physiologic intercellular messenger to promote platelet aggregation and the release reaction. The activity is formed by intact cells after physiologic stimulation or can be generated from platelet membrane fractions after combination with arachidonate. In the present investigation, small amounts of polyunsaturated fatty acids added to an incubation mixture of platelet microsomes and arachidonate were found to significantly inhibit subsequent platelet aggregation. Saturated and mono-unsaturated fatty acids in the same concentrations were without effect. However, in higher concentrations mono-unsaturated fatty acids were found to be inhibitory and stearic acid was found to enhance subsequent platelet aggregation. The inhibition caused by the polyunsaturated fatty acid, linoleate, was shown to be the result of an effect on the production of LASS through an interaction with the platelet enzyme responsible for conversion of arachidonate to LASS. In contrast, stearic acid was found to enhance platelet aggregation by acting on the platelets and not directly on LASS production. The results suggest that small changes in the fatty acid composition of platelet phospholipids could significantly influence platelet reactivity.


Defective leukocyte metabolism in human cobalamin deficiency: impaired propionate oxidation and serine biosynthesis reversible by cyanocobalamin therapy. Biochemical disturbances common to vitamin B12 and folate deficiency were investigated in leukocytes from patients with cobalamin deficiency. The investigations focused on the only two human metabolic pathways known to require vitamin B12. In the propionate pathway, deoxyadenosylcobalamin is required for isomerization of methylmalonyl-CoA to succinyl-CoA. Leukocyte oxidation of 14C-propionate to 14CO2 was markedly decreased in 9 patients with Addisonian cobalamin deficiency and 2 patients with low serum cobalamin associated with folate deficiency, whereas 14C-succinate oxidation was normal. Three of the Addisonian patients had only minimal anemia. Within 4 days after one injection of 1,000 mug of cyanocobalamin, in 7 out of 8 patients studied, leukocyte propionate oxidation increased to normal levels. In folate-mediated one-carbon metabolism, as measured by serine biosynthesis from formate, methylcobalamin is required for conversion of methyl-folate to tetrahydrofolate. Leukocyte formation of 14C-serine from 14C-formate was significantly depressed in 5 patients with low serum cobalamin, little or no anemia, and only marginally low total red cell folate, the low serum cobalamin in 2 of these patients was associated with folate deficiency. After 1,000 mug of cyanocobalamin, in 2 of 3 patients, leukocyte serine biosynthesis increased to the normal range. These observations demonstrated that these two metabolic pathways in leukocytes were sensitive to cobalamin deficiency, and responsive to cobalamin therapy. Although there was no correlation between either of these metabolic activities and the serum cobalamin, red cell folate, or hematocrit, there was a striking correlation between impairment of leukocyte propionate oxidation and of leukocyte serine biosynthesis in 5 patients who were minimally anemic. The remarkably close correspondence between effects of low cobalamin on these two metabolic pathways, in nonanemic patients, must be a direct consequence of their common requirements for a cobalamin co-enzyme. These findings emphasize the importance of cobalamin in folate metabolism, and are consistent with the hypothesis that folate is "trapped" as methyl-folate in cobalamin deficiency, but do not exclude the possibility that this "trapping" is caused by a third metabolic function of cobalamin which might mediate transport of folate into cells.


Failure of water immersion to influence parathyroid hormone secretion and renal phosphate handling in normal man. Previous studies from this laboratory have demonstrated that the redistribution of blood volume and concomitant relative central hypervolemia induced by water immersion to the neck (NI) results in a significant natriuresis which is quantitatively identical to that induced by the acute administration of 2 L. of saline. Since the central hypervolemia induced by NI occurs without concomitant alterations in serum ionized calcium concentration (Ca++), the NI model was utilized to assess the role of volume in the regulation of PTH secretion in man. Seven normal subjects were studied following 11 hours of dehydration on two occasions, control and NI. The conditions of seated posture and time of day were identical. Blood for ionized calcium and PTH was obtained at 30-minute intervals for 6 hours. NI resulted in a significant increase in UNaV from a prestudy value of 78 +/- 12 (S.E.M.) to 222 +/- 20 muEq per minute (p less than 0.001). Concomitantly, Ca++ remained constant, ranging between 4.57 to 4.71 mg. per cent. Despite the volume-induced natriuresis, PTH was not altered throughout 5 hours of NI, ranging from 36 +/- 7 to 45 +/- 5 mul-Eq. per milliliter. Phosphate excretion remained constant. These data indicate that central volume expansion does not alter PTH in normal man when the variables of ionized calcium, posture, and time of day are controlled. Furthermore, the current demonstration of the absence of phosphaturia during immersion despite the probability that the distal delivery of phosphate was enhanced, permits consideration of the possibility that the concept of a distal tubular reabsorptive site for phosphate may be applicable to man.


Hormonal therapy and alteration of lymphocyte proliferation. Suppression of the blastogenic response of phytohemagglutinin-stimulated peripheral blood lymphocytes in autologous serum obtained from patients with prostatic cancer. Peyronie's disease, and transsexuals following estrogen therapy has been observed. Observations of suppression of blastogenesis in patients with prostatic cancer and without malignancy receiving estrogen suggests that this suppression would appear not to be associated with a particular pathologic state or malignancy per se but, rather, the given mode of therapy. The relevance of these observations to the effect of estrogen on cell-mediated responsiveness in patients with hormonally dependent tumors and to the suggested association between uterine cancer and prolonged administration of diethylstilbesterol and the development of vaginal tumors in offspring found in association with material ingestion during pregnancy remains speculative.


The effects of acute infectious hepatitis and cirrhosis of the liver on the nonerythropoietic component of early bilirubin. The nonerythropoietic component of early labeled bilirubin in plasma and bile was studied in 7 patients with acute infectious hepatitis during the late convalescent stage, in 13 patients with Laennec-type liver cirrhosis, and in 7 control subjects after intravenous injection of a tracer dose (2.5 muCi) of 4-14C-delta-aminolevulinic acid (14C-deltaALA). All subjects were examined during the nonicteric stage. In control subjects, the mean cumulative radioactivity readings in 4 hours were 29.6 +/- 4.7 X 10(3) d.p.m. per milligram times 4 hours in plasma and 27.0 +/- 1.2 X 10(3) d.p.m. per milligram times 4 hours in bile. In acute infectious hepatitis patients, the mean cumulative radioactivity readings for both plasma and bile in 4 hours were approximately twice that found in control subjects. In mild cirrhotic patients with enlarged liver on scintigram, the mean cumulative radioactivity readings for both plasma and bile were approximately 1.4 times that in control subjects. In patients with more advanced cirrhosis and markedly small livers on scintigram, the mean cumulative radioactivity readings for both plasma and bile were as low as approximately 0.5 that of control subjects. These findings seem to indicate the important role of the liver in the production of the nonerythropoietic component of early bilirubin in man.


Exaggerated natriuresis in the conscious spontaneously hypertensive rat. In response to an acute saline load, many patients with essential hypertension exhibit an exaggerated natriuresis relative to normotensive controls. In the present study, the urinary responses of conscious,Okamoto-strain, spontaneously hypertensive rats (SHR), and Wistar-Kyoto strain normotensive rats (NTR) to an acute saline load were evaluated to determine if a similar exaggerated natruiresis exists in this form of hypertension. Twelve rats of each strain per group (12 weeks of age) were housed in metabolism cages for 1 week. Systolic blood pressures (tail cuff) were significantly different (206+/- 9 mm. Hg in SHR and 135 +/- 3 mm. Hg in NTR). After a 4-hour control urine collection, 6 ml. of 0.9 per cent sodium chloride were given by gavage. Urine was collected again for 2 hours. Control urinary excretions of sodium, potassium, and creatinine in SHR and NTR were 11.2 +/- 4.8 muEq per hour, 50.1 +/- 7.6 muEq per hour, and 39.9 +/- 5.5 mg. per hour in SHR, and 13.8 +/- 2.4 muEq per hour, 34.9 +/- 5.5 muEq per hour, and 37.5 +/- 7.1 mg. per hour in NTR, respectively. The respective control values for sodium, potassium, and creatinine excretion in the two groups were not significantly different. Following the saline load, sodium and creatinine excretion rates were significantly elevated in both groups of rats. However, the increase in sodium excretion in SHR (60.8 +/- 7.2 MUEq per hour) was more than double and significantly different from that of the NTR (26.6 +/- 3.7 muEq per hour). In contrast, the increments in creatinine excretion in the two groups of rats were not significantly different from each other. In the NTS, urinary potassium excretion was significantly elevated (59.0 +/- 7.9 muEq per hour) whereas in SHR it was not significantly altered (12.0 +/- 8.8 muEq per hour). The change in urinary creatinine excretion as an index of change in glomerular filtration rate suggests that the greater increase in sodium excretion by the SHR was the result of decreased fractional reabsorption of sodium and not the result of a greater increase in glomerular filtration rate. The exaggerated natriuretic response to salt loading in SHR resembles that in hypertensive man except that in SHR, a simultaneous kaliuretic response is absent.


Effect of chenodeoxycholic acid and phenobarbital on the rate-limiting enzymes of hepatic cholesterol and bile acid synthesis in patients with gallstones. The effects of chenodeoxycholic (CDC), 750 mg. per day, phenobarbital (PB), 90 or 180 mg., combined (CDC + PB), and placebo on biliary lipid composition and on the rate-limiting enzymes of hepatic cholesterol synthesis (HMG-CoA reductase) and bile acid synthesis (cholesterol 7alpha-hydroxylase) were studied. Percutaneous liver biopsies were performed after 6 months of therapy in 4 patients from each group participating in a double-blind study of gallstone dissolution. The enzyme activities were also assayed in liver obtained at laparotomy in 7 untreated gallstone patients and 4 without gallstones. 7alpha,12alpha-Dihydroxycholest-4-en-3-one-12alpha-hydroxylase, an enzyme leading to cholic acid synthesis, was determined in 4 untreated gallstone patients and 4 without gallstones. Untreated gallstone patients had 35 per cent greater HMG-CoA reductase (p less than 0.01), 37 per cent less 7alpha-hydroxylase (p less than 0.01), and 40 per cent less 12alpha-hydroxylase (p less than 0.01) than patients without gallstones. CDC, PB, and both increased biliary CDC and decreased the lithogenic index significantly (p less than 0.01) but saturated bile persisted with PB. CDC decreased HMG-CoA reductase 40 per cent (p less than 0.01) and 7alpha-hydroxylase 47 per cent (p less than 0.01). PB increased HMG-CoA reductase 112 per cent (p less than 0.01) and 7alpha-hydroxylase 20 per cent (p less than 0.01). The combination of CDC and PB increased HMGCoA reductase 40 per cent (p less than 0.01) and had no effect on 7alpha-hydroxylase. In conclusion, CDC induced desaturation of bile while decreasing HMG-CoA reductase and increasing CDC in bile. PB reduced the saturation less effectively than CDC; it increased 7alpha-hydroxylase but also increased HMG-CoA reductase.


Artifacts in the radioimmunoassay of peptide hormones in gastric and duodenal secretions. Observations of apparently high concentrations of gastrin and insulin in gastric and duodenal secretions as determined by radioimmunoassay may arise from artifacts caused by proteolytic enzymes. The double-antibody and charcoal separation techniques as commonly employed fail to recognize extensive proteolytic damage to labeled antigen and to antibody.


Sensitivity to radiation and cycle-active drugs as a function of stem cell compartment repletion. We have studied the sensitivity of normal mouse hemopoietic tissue to radiation and cycle-active drugs in relation to stem cell compartment repletion. Recovery of erythropoiesis in endogenous spleen colonies, blood reticulocytes, and 30-day survivals were determined in mice after an initial large dose of partial-body irradiation. We found that the normal stem cell compartment is more sensitive to cycle-independent modes of therapy, like radiation and cyclophosphamide, than it is to cycle-active agents like cytosine arabinoside and methotrexate. The depleted stem cell compartment exhibits marked sensitivity to cycle-independent agents but less to cycle-active agents, which, however, suppress its recovery more than they do the normal. The overshoot phase of recovery is relatively resistant to either cycle-independent or cycle-active agents. A reticulocytosis following a reticulocytopenia signals the overshoot phase of stem cell compartment recovery and relatively increased resistance. These findings may prove useful in designing chemotherapy regimens and in anticipating marrow recovery in planning for supportive care in patients with neoplastic disease.


Kininogen deficiency in Fitzgerald trait: role of high molecular weight kininogen in clotting and fibrinolysis. Plasma from an asymptomatic person with defects in blood coagulation, release of kinin, and evolution of fibrinolytic activity upon contact with a foreign surface was deficient in kininogen. The coagulation defect was identified as "Fitzgerald trait." A preparation of high molecular weight kininogen repaired the defects in clotting, kinin release and fibrinolysis, but a preparation of low molecular weight kininogen did not. Therefore, Fitzgerald factor appears to be a high molecular weight kininogen. The site of action of the kininogen appears to be after that of activated Hageman factor and kallikrein in the generation of clot-promoting activity through activation of plasma thromboplastin antecedent (PTA).


A heterologous radioimmunoassay for arginine vasopressin. A sensitive and specific radioimmunoassay for arginine vasopressin (AVP) was developed utilizing the antisera against lysine vasopressin (LVP) in combination with a labeled AVP. The assay employs an acetone extraction procedure and detects as little as 0.8 pg. per milliliter of AVP in human plasma. In normal subjects the mean (+/- S.D.) plasma concentration of AVP was 4.9 +/- 1.2 pg. per mililiter after fluid deprivation and 1.2 +/- 0.4 pg. per milliliter after water loading. Plasma AVP levels correlated significantly with plasma osmolalities. In four patients with diabetes insipidus, plasma AVP concentrations ranged from less than 0.8 to 1.2 pg. per milliliter, whereas six patients with the syndrome of inappropriate ADH secretion showed plasma levels of AVP which correspond to those of the dehydrated state in normal subjects or greater, although plasma osmolalities were low in all cases. It was concluded that the present radioimmunoassay method for AVP provides a useful way of assessing neurohypophyseal function in man.


Direct spectrofluorometry of diluted erythrocytes and plasma: a rapid diagnostic method in primary and secondary porphyrinemias. Primary and secondary porphyrinemias are characterized by the presence of several porphyrin species in erythrocytes and/or plasma. Acid solvent extraction of porphyrins from natural ligands can preclude observation of significant spectral differences apparent on direct spectrofluorometric examination of porphyrin complexes in tissue. Erythrocytes and hemoglobin-free plasma obtained from 13 patients with erythropoietic, 9 with lead intoxication, 5 with porphyria cutanea tarda, 3 with iron deficiency anemia, 1 with erythropoietic porphyria, and 10 normal persons were simply diluted and analyzed spectrofluorometrically. Characteristic porphyrin-ligand spectral patterns were consistently obtained for each group of specimens, allowing diagnostic application of the data.


Anterior conduction delay: a possible cause for prominent anterior QRS forces. Prominently anterior QRS forces often present a diagnostic dilemma. Frequently, right ventricular forces may be eliminated on clinical grounds and dorsal infarction is therefore suspected, especially in a clinical setting of coronary artery disease. In five such patients studied angiographically, the coronary disease was concentrated in the left anterior descending artery and the ventricular dysfunction confined to the anterior wall of the left ventricle. In a sixth case, the prominent anterior forces were observed intermittently together with left anterior hemiblock. These observations, in addition to serial studies following surgery, strongly suggest that the mechanism for prominent anterior QRS forces in these cases is conduction delay in an anterior division of the left bundle branch system.


Effect of cold isotonic glucose infusion on A-V nodal conduction. His bundle electrograms were performed in eight patients with organic heart disease. Recordings were made at various rates utilizing right atrial pacing. A solution of 100 cc of iced cold 5% glucose and water was infused through a cardiac catheter placed at the level of the tricuspid valve. The P-A, A-H, H-Q and H-S intervals were determined before and immediately after the cold water infusion. A significant prolongation of the A-H interval occurred with negligible effects on the P-A, H-Q and H-S intervals. At the atrial pacing rate of 100/min the average A-H interval increased from the control value of 116 to 125 msec after the infusion (P less than 0.02); at the pacing rate of 140/min, the A-H interval increased from 147 to 158 msec (P less than 0.01). This represents an impairment in conduction through the atrioventricular node.


Prediction of pulmonary arterial pressures from electrovectorcardiographic data in pediatric patients with rheumatic mitral valve disease. The Frank vectorcardiogram (VCG) and the electrocardiogram (ECG) were used to predict pulmonary arterial pressures in 30 pediatric patients with rheumatic mitral valve disease. The patients' ages ranged from eight to 14 1/2 years. Eleven were male and 19 were female. Sixteen had mitral stenosis, eight had mitral regurgitation and six had mitral stenosis and regurgitation. Mean pulmonary arterial wedge pressure ranged from 5-32 mmHg. All patients underwent complete catheterization and angiocardiographic study. None had significant gradient across the right ventricular outflow tract. Right maximum spatial vector (Rmsv) was calculated using Frank VCG.R in V1 and S in V5 of ECG were also measured. Rmsv, RV1, and SV5 were correlated with pulmonary arterial pressures (systolic, diastolic and mean). Pulmonary artery systolic pressure and Rmsv bear the best correlation (r=0.773). The correlation coefficient for pulmonary artery diastolic pressure and Rmsv was 0.698. Rmsv as calculated from Frank VCG is useful in prediction of pulmonary arterial pressures (systolic and diastolic) in pediatric patients with rheumatic mitral valve disease.


A simple procedure for positioning precordial ECG and VCG electrodes using an electrode locator. Methodological differences in the placement of precordial ECG electrodes are a major problem in multicenter clinical trials and epidemiological studies. Trend analysis and realization of the full potential of computer programs for serial comparison demands reduction of technical sources of variation in the electrocardiogram (ECG) and vectorcardiogram (VCG) and particularly errors in locating ECG electrodes. The ECG electrodes locator described here reduces a major precordial uncertainty in the identification of the midclavicular and the anterior axillary lines. It simplifies positioning and provides a numeric record of the key precordial electrode positions, facilitating control of electrode placement errors in serial recordings.


Evidence for local cardiac effects in electrocardiograms from a large electrode array. The sensitivity of an extensive electrocardiographic examination to local cardiac regions was evaluated by comparison with findings in vectorcardiographic leads. Evaluation consisted of comparing QRS areas during different ventricular activation patterns. The ratio of areas from the vector-cardiogram (VCG) and extensive electrode systems differed by 15 to 23 percent, suggesting at least this degree of local cardiac influence on the extensive examination.


Corrected transposition of the great vessels: a vectorcardiographic study. The vectorcardiograms (VCGs) of two groups of patients with corrected transposition of the great vessels (CTGV) were studied; the first, group A, included 17 patients with CTGV in "situs solitus,", characterized by leftward orientation of the cardiac apex; the second group, group B, with three patients, presented CTGV in "situs solitus" and apex to the right. All cases had one or more associated defects: ventricular septal defect, atrial septal defect, pulmonic stenosis or tricuspid insufficiency.


Wandering atrial pacemaker (prevalence in French hornists). Continuous electrocardiographic recordings were obtained in a group of French horn players during performance of identical pieces of music. Half of the musicians developed wandering atrial pacemaker. One example is illustrated. The causative mechanism is briefly discussed. This may represent an "occupational" hazard.