The role of the medullary collecting ducts in postobstructive diuresis.

Medullary collecting duct function was studied by direct microcatheterization techniques in rats undergoing postobstructive diuresis. Significant net addition of water and sodium to the duct was demonstrated during postobstructive diuresis after relief of 24-h bilateral ureteral ligation. This striking abnormality in function was associated with reduced delivery of sodium and water to the collecting duct compared to sham-operated controls. To examine the role of circulating factors in this phenomenon, another group of rats was studied that underwent 24 h of total urine reinfusion into the femoral vein. Natriuresis and diuresis were similar to the postobstructive group, but absolute collecting duct reabsorption of sodium and water was normal. The natriuresis and diuresis in rats with urine reinfusion resulted from increased delivery of fluid and sodium to the medullary collecting duct. A third group of rats was studied with 24-h unilateral ureteral ligation as well as urine reinfusion from the contralateral normal kidney. Without urine reinfusion there was no diuresis-natriuresis but with urine reinfusion the diuresis and natriuresis after relief of unilateral obstruction was similar to that after relief of bilateral obstruction. Moreover, net addition of sodium and no significant water reabsorption were demonstrated in the medullary collecting duct of such animals. The results indicate that (a) the medullary collecting duct is the critical nephron segment affected by ureteral obstruction, since postobstructive diuresis occurred despite reduced delivery of fluid from the more proximal nephron; (b) the net addition of sodium to the medullary collecting duct observed during postobstructive diuresis is probably a direct effect of obstruction, since it was found during postobstructive diuresis after relief of bilateral or unilateral ureteral ligation, but not with urine reinfusion alone; and (c) blood-borne factors are important in the development of postobstructive natriuresis and diuresis, and probably act by increasing the fraction of filtered sodium and water delivered from the proximal and distal tubule to the collecting duct.


Functional adaptation of nephrons in dogs with acute progressing to chronic experimental glomerulonephritis.

Although a diminished fractional excretion of sodium (FENa) is the hallmark of acute proliferative glomerulonephritis (APGN), an enhanced natriuresis per glomerular filtration rate (GFR) in the chronic phases of this disease has been reported. We studied this adaptive response utilizing two different split-bladder dog models with unilateral, and a third group of dogs with bilateral Masugi's nephritis. Group I. Six dogs with unilateral nonaccelerated APGN studied a mean of 6 days after induction had a mean base-line APGN/intact kidney GFR of 31/50 ml/min (P less than 0.005) and FENa of 0.2/0.75% (P less than 0.005). Acute volume expansion caused a smaller absolute increase in FENa from the APGN kidney, 1.6%, than from the intact kidney, 4.0%, (P less than 0.01). Maximum tubular secretion of rho-aminohippuric acid/GFR (TmPAH/GFR) measured in three dogs was higher in the APGN kidney than intact kidney, 13.1 vs. 9.3 mg/dl. Subsequent studies on three of the six dogs when the disease had become chronic demonstrated a reversal in the pattern of sodium excretion in response to volume expansion. Group II. Six dogs with accelerated unilateral APGN (dogs presensitized to antibody source) studied a mean of 5 days after induction had a mean base-line APGN/intact kidney GFR of 16/57 ml/min and FENa of 0.22/0.12% (P less than 0.1). Contrary to group I, volume expansion caused a greater absolute increase in FENa from the APGN kidney, 5.8%, than from the intact kidney, 2.9% (P less than 0.05). TmPAH/GFR studied in four dogs was similar for both kidneys, 17.9 and 18.5 mg/dl for the APGN kidney and intact kidney, respectively. Group III. Sequential studies were performed on seven dogs with bilateral nonaccelerated APGN. Initially each demonstrated sodium retention and a smaller absolute increase in FENa in response to volume expansion compared to a predisease control study. With disease progression, volume expansion induced a greater absolute increase in FENa than in the control study. We concluded that (a) the fractional excretion of sodium from the APGN kidney will be less or greater than the contralateral intact kidney or control study depending on the severity and/or chronicity of the disease, possibly as the result of morphologic alterations; (b) the degree of extracellular fluid volume expansion is an important variable influencing similarity of glomerulotubular balance between the APGN and contralateral intact kidney; and (c) the "intact nephron hypothesis" applies in a limited fashion to kidneys with APGN in the absence of volume expansion just as it does for kidneys with chronic glomerulonephritis or pyelonephritis.


Hypoxanthine phosphoribosyltransferase activity in intact fibroblasts from patients with X-linked hyperuricemia.

Discordance between clinical phenotype and the level of a mutant enzyme activity may reflect differences between enzyme function in vivo and that measured by the customary enzyme assays on cell extracts. In the present study, the conversion of hypoxanthine to phosphorylated products was measured in intact skin fibroblasts and in cell extracts from seven patients with mutant hypoxanthine-guanine phosphoribosyltransferase (HPRT) and six control subjects. The patient's phenotypes ranged from asymptomatic hyperuricemia to the Lesch-Nyhan syndrome. Although there was a general correlation between the HPRT activity in cell extracts assayed by the usual methods and the function of the purine salvage pathway in patients, as reflected by urinary oxypurine excretion, there were notable exceptions. A more accurate appraisal of the functioning of the pathway at the cellular level is achieved by measuring the conversion of substrate to product in the intact cell at physiological concentrations of substrates, activators, and product and metabolite inhibitors, and in a physiological ionic environment. In one of the seven patients, the standard enzyme assay indicated normal function, whereas measurements in the intact cell exposed severe dysfunction of the salvage system. In another, the standard assay suggested a severe deficiency not evident in the intact cell or in the patient.


Hereditary deficiency of the fifth component of complement in man. I. Clinical, immunochemical, and family studies.

The first recognized human kindred with hereditary deficiency of the fifth component of complement (C5) is described. The proband, a 20-year-old black female with systemic lupus erythematosus since age 11, lacked serum hemolytic complement activity, even during remission. C5 was undetectable in her serum by both immunodiffusion and hemolytic assays. Other complement components were normal during remission of lupus, but C1, C4, C2, and C3 levels fell during exacerbations. A younger half-sister, who had no underlying disease, was also found to lack immunochemically detectable C5. By hemolytic assay, she exhibited 1-2% of the normal serum C5 level and normal concentrations of other complement components. C5 levels of other family members were either normal or approximately half-normal, consistent with autosomal codominant inheritance of the gene determining C5 deficiency. Normal hemolytic titers were restored to both homozygous C5-deficient (C5D) sera by addition of highly purified human C5. In specific C5 titrations, however, it was noted that when limited amounts of C5 were assayed in the presence of low dilutions of either C5D serum, curving rather than linear dose-response plots were consistently obtained, suggesting some inhibitory effect. Further studies suggested that low dilutions of C5D serum contain a factor (or factors) interfering at some step in the hemolytic assay of C5, rather than a true C5 inhibitor or inactivator. Of clinical interest are (a) the documentation of membranous glomerulonephritis, vasculitis, and arthritis in an individual lacking C5 (and its biologic functions), and (b) a remarkable propensity to bacterial infections in the proband, even during periods of low-dose or alternate-day corticosteroid therapy. Other observations indicate that the C5D state is compatible with normal coagulation function and the capacity to mount a neutrophilic leukocytosis during pyogenic infection.


Mechanism of action of pentagastrin on the lower esophageal sphincter.

The effects of pentagastrin on lower esophageal sphincter (LES) pressure has been studied in trained, unanesthetized dogs. LES pressure was monitored by an infusion manometric technique. Increasing doses of pentagastrin up to 3 mug/kg given as an i.v. bolus resulted in increasing rises in LES pressure; larger doses resulted in a lesser effect of shorter duration. Increasing i.v. boluses of methacholine produced greater increases in LES pressure up to a maximum of 5 mug/kg; higher doses had similar effects. Atropine (50-100 mug/kg) slightly diminished the response of the LES to 2 or 6 mug/kg of pentagastrin. In large doses (500-2,000 mug/kg), atropine did not diminish the response to pentagastrin and prolonged the response of 6 mug/kg pentagastrin. Hexamethonium (2 mg/kg i.v.) depressed the peak response to 3 mug/kg pentagastrin slightly but the response to 6 mug/kg was increased and prolonged. Propranolol (2 mg/kg i.v.) significantly prolonged the effect of 6 mug/kg pentagastrin on the LES. We conclude that the stimulatory effect of pentagastrin is mainly due to a direct action on the LES. A lesser stimulatory effect is due to an action on preganglionic cholinergic neurons. Large doses of pentagastrin have both stimulatory and inhibitory effects. The inhibitory effect is mediated at least in part via preganglionic neurons acting through adrenergic receptors. Ganglionic transmission of the effect may be through muscarinic as well as nicotinic receptors.


The biosynthesis of human hemoglobin A1c. Slow glycosylation of hemoglobin in vivo.

Hemoglobin A1c, the most abundant minor hemoglobin component in human erythrocytes, is formed by the condensation of glucose with the N-terminal amino groups of the beta-chains of Hb A. The biosynthesis of this glycosylated hemoglobin was studied in vitro by incubating suspensions of reticulocytes and bone marrow cells with [3H]leucine or 59Fe-bound transferrin. In all experiments, the specific activity of Hb A1c was significantly lower than that of Hb A, suggesting that the formation of Hb A1c is a posttranslational modification. The formation of Hb A1c in vivo was determined in two individuals who were given an infusion of 59Fe-labeled transferrin. As expected, the specific activity of Hb A rose promptly to a maximum during the 1st week and remained nearly constant thereafter. In contrast, the specific activity of Hb A1c and also of Hbs A1a and A1b rose slowly, reaching that of Hb A by about day 60. These results indicate that Hb A1c is slowly formed during the 120-day life-span of the erythrocyte, probably by a nonenzymatic process. Patients with shortened erythrocyte life-span due to hemolysis had markedly decreased levels of Hb A1c.


Effect of growth hormone-release inhibiting hormone on hormones stimulating exocrine pancreatic secretion.

The nature and extent of growth hormone-release inhibiting hormone (GH-RIH, somatostatin)-induced inhibition of pancreatic secretion of bicarbonate and protein, an index of enzyme secretion, were studied by administration of exogenous secretin or cholecystokinin (CCK) and of a number of stimulants for endogenous release of these hormones in fasted pancreatic fistula dogs with and without an infusion of GH-RIH. The results of this study show that GH-RIH inhibits the pancreatic fluid and bicarbonate secretion induced by duodenal acidification and exogenous secretion. The kinetic analysis shows that the interaction between GH-RIH and secretin affecting pancreatic bicarbonate secretion possesses the characteristics of competitive inhibition. GH-RIH does not change the pancreatic protein response to exogenous CCK, but profoundly inhibits pancreatic response to a variety of the endogenous stimulants of CCK release, including duodenal perfusion of sodium oleate, amino acid mixture, or feeding of a peptone meal. We conclude that GH-RIH is a very potent inhibitor of the endogenous release of CCK from the intestinal mucosa and inhibits competitively the action of secretin but not CCK on the exocrine pancreatic secretion.


Fibrinopeptide A in plasma of normal subjects and patients with disseminated intravascular coagulation and systemic lupus erythematosus.

A radioimmunoassay for fibrinopeptide A (FPA) has been developed. This assay uses rabbit antibodies induced by injection of native FPA-human serum albumin conjugates and 125I introduced into tyrosine-FPA synthesized in out laboratory. Plasma FPA is separated from fibrinogen by TCA extraction. The assay is capable of detecting as little as 50 pg/ml of FPA. In 20 normal donors this assay revealed a mean concentration of 0.9 ng/ml (0.3 SD). In five patients with disseminated intravascular coagulation, FPA concentrations ranged from 13.0 to 346 ng/ml. Two groups of patients with systemic lupus erythematosus (SLE) whose disease had achieved complete remission were studied; one consisted of four patients with no history of lupus nephritis and another with a history of nephritis. Mean FPA concentrations of 1.5 ng/ml (range, 0.7-1.8 ng/ml) and 2.7 ng/ml (range, 1.1-5.6 ng/ml) were found in these two groups, respectively. Another group of nine patients with active SLE, but without evidence of lupus nephritis, had a mean FPA concentration of 4.5 ng/ml (range, 2.4-7.8 ng/ml). Finally, a group of seven patients with active SLE, including active nephritis, had a mean FPA concentration of 10.2 ng/ml (range, 5.3-17.0 ng/ml). A positive correlation was found between the concentration of plasma FPA and serum DNA-binding activity and an inverse correlation was found between plasma FPA and the concentration of serum C3. No correlation existed between plasma FPA and concentration of serum creatinine. Several possibilities for the origin of plasma FPA in patients with SLE were considered; at present it seems most likely that FPA arises through the action of thrombin on fibrinogen.


A comparative analysis of the C1-binding ability of fragments derived from complement-fixing and noncomplement-fixing IgM proteins.

The purpose of this study was to examine the molecular parameters necessary for initiation of complement fixation by IgM proteins. To determine why some IgM molecules are capable of complement fixation while others are not, several different Waldenström IgM proteins were examined for their ability to fix total hemolytic complement in the CH(50) assay. Subsequently, the C1 fixing ability of a 56-residue fragment derived from the Cmu4 domain of each of these IgM molecules was studied with C1 fixation assay. One of the three Waldenström IgM proteins (Gr) used in the present study was found unable to consume complement in a CH(50) assay when tested at the same concentration as the two complement-consuming IgM molecules (Dau and Bus). However, when the 56-residue C(H)4 fragment from the Cmu4 domain of each IgM molecule was tested for C1-fixing ability, all three were found to bind C1. On the basis of these observations, it is proposed that a C1 binding site exists within the Cmu4 domain of both complement-fixing and noncomplement-fixing IgM molecules. Presumably, the latter molecules are unable to interact in their native state with C1 in the manner required for initiation of the classical complement pathway, possibly due to the configurational inaccessibility of the entire C1 binding site.


Metabolism of [1-(14)C] and [2-(14)C] leucine in cultured skin fibroblasts from patients with isovaleric acidemia. Characterization of metabolic defects.

Leucine metabolism in cultured skin fibroblasts from patients with isovaleric acidemia was compared with that in normal fibroblasts and in cells from patients with maple syrup urine disease using [1-(14)C] and [2-(14)C] leucine as substrates. Inhibitory effects of methylenecyclopropylacetic acid on leucine metabolism in normal cells were also investigated. Production of 14CO2 from [2-(14)C] leucine was very reduced (96-99%) in both types of mutant cells. Radioactive isovaleric acid accumulated in assay media with isovaleric acidemia cells but not in those with maple syrup urine disease cells. Unexpectedly, 14CO2 production from [1-(14)C] leucine was partially depressed (80%) in isovaleric acidemia cells whereas in maple syrup urine disease cells it was strongly depressed (99%) as expected. These two mutant cells were clearly distinguished by detection of 14C-isovaleric acid accumulation after incubation with [2-(14)C] leucine. A pattern of inhibition of leucine oxidation similar to that seen in isovaleric acidemia cells was induced in normal cells by the addition of 0.7 mM methylenecyclopropylacetic acid to the assay medium. The partial inhibition of [1-(14)C] leucine oxidation seen in isovaleric acidemia cells and also in normal cells in the presence of the inhibitor appears to be, at least in part, due to an accumulation of isovalerate in the cells. Isovaleric acid (5-10) mM) inhibited [1-(14)C] leucine oxidation 32-68% when added to the assay medium with normal cells. Addition of flavin adenine dinucleoside to culture medium or assay medium or both did not restore oxidation of either leucine substrate in isovaleric acidemia cells.


Cystinosis. Intracellular cystine depletion by aminothiols in vitro and in vivo.

Certain aminothiols rapidly deplete cultured cystinotic skin fibroblasts of their abnormally high free (nonprotein) cystine pool. The free cystine content of these cells if reduced by over 90% in 1 h with 0.1 mM cysteamine. This is more rapid than previously known methods of removing free cystine from cystinotic fibroblasts. The disulfide, cystamine, is also able to deplete cystinotic cells of free cystine. A patient with nephropathic cystinosis and end-stage renal disease was treated with cysteamine, both intravenously and orally. Both methods of administration rapidly lowered the free cystine content of the patient's peripheral leukocytes. Study of the patient's urinary sulfur excretion did not conclusively determine the effect of this therapy on the total body cystine pool. Her renal status remained at end stage after 1 mo of oral cysteamine, when an episode of grand mal seizures prompted cessation of the study. Determination of the proper place of aminothiol therapy in this disease will depend upon further clinical trial with patients whose kidney function has not deteriorated to the point of irreversible change, accompanied by careful monitoring of plasma aminothiol levels.


Physical chemistry of the lipids of human atherosclerotic lesions. Demonstration of a lesion intermediate between fatty streaks and advanced plaques.

95 individual human atherosclerotic lesions from 26 persons were classified into three groups under the dissecting microscope: fatty streaks, fibrous plaques, and gruel (atheromatous) plaques. Each lesion was isolated by microdissection, its lipid composition determined by chromatography, and the physical states of the lipids identified by polarizing microscopy and in some cases by X-ray diffraction. The composition of each lesion was plotted on the in vitro phase diagram of the major lipids of plaques: cholesterol, cholesterol ester, and phospholipid. The observed physical states were compared with those predicted by the location of the lipid composition on the phase diagram. The most severe lesions (gruel plaques) had an average lipid composition of cholesterol 31.5+/-1.9%, cholesterol ester 47.2+/-2.3%, and phospholipid 15.3+/-0.5%. Their compositions fell within the three-phase zone of the phase diagram, predicting the lipids to be separated into a cholesterol crystal phase, a cholesterol ester oily phase and a phospholipid liquid crystalline phase. In addition to the phospholipid liquid crystalline phase of membranes and myelin-like figures demonstrable by electron microscopy, polarizing microscopy revealed the other two predicted phases, isotropic cholesterol ester-rich droplets and cholesterol crystals. X-ray diffraction studies verified the identity of the crystals as cholesterol monohydrate. Fibrous plaques also had an average lipid composition within the three-phase zone of the phase diagram. Polarizing microscopy revealed the presence of cholesterol monohydrate crystals and lipid droplets in all of these lesions; the droplets were predominately isotropic in 28 of the 31 fibrous plaques. Although these lesions had less free cholesterol and more cholesterol ester than gruel plaques, they were otherwise similar. Fatty streaks had compositions within both the two- and three-phase zones of the phase diagram. Compared with gruel plaques, the fatty streaks within the two-phase zone, defined as "ordinary," had more cholesterol ester, less free cholesterol, a higher cholesteryl oleate/cholesteryl linoleate ratio, a lower sphingomyelin/lecithin ratio, more anisotropic lipid droplets, and rare or no cholesterol crystals. Those lesions within the three-phase zone had many chemical and physical features intermediate between ordinary fatty streaks and gruel plaques. Moreover, 68% of these "intermediate" lesions had no cholesterol crystals by polarizing microscopy in spite of their compositions being within the three-phase zone, indicating the cholesterol ester oily phase or the phospholipid phase or both were supersaturated with cholesterol. Identification of this group of intermediate lesions provides evidence that some fatty streaks may be precursors of advanced plaques.


Interactions of acetylcholine and epinephrine on the dynamics of insulin release in vitro.

An in vitro system for perifusion of rat pancreatic islets has been utilized to define the effects of epinephrine on acetylcholine-induced insulin release over varying concentrations of the two agents. Perifusion of islets with epinephrine before challenge with acetycholine produced marked enhancement of both phases of cholinergically induced insulin release; enhancement of the first phase being more marked with increase in acetylcholine concentration and the converse being observed with the second phase. Perifusion of islets with epinephrine during stimulation with acetylcholine produced inhibition of insulin release, an effect dependent upon the concentration of epinephrine and of acetylcholine. There was an order of difference in the acetycholine concentration needed to overcome significant epinephrine-mediated inhibition of the first phase of insulin release (5 X 10(-4) mug/ml) and that needed to overcome inhibition of the second phase (5 X 10(-3) mug/ml). Comparison of the effects of various concentrations of epinephrine on glucose- and acetyl-choline-induced insulin release revealed that epinephrine was a less potent inhibitor of the first phase of acetylcholine-induced insulin release than of the first phase of glucose-induced insulin release. These data provide some insight into the potential interactions between cholinergic and adrenergic autonomic systems in modifying insulin release.


Lymphokine stimulation of collagen accumulation.

Lymphokine-rich supernates from normal human peripheral blood mononuclear cells, stimulated by the mitogen phytohemagglutinin, have been shown to cause enhanced collagen accumulation by human embryonic lung fibroblasts (WI-38), as measured by hydroxyproline content of fibroblast monolayers, [14C] proline incorporation into soluble collagen and collagenase release of radioactivity in supernates and monolayers of cultures incubated with [14C] proline. This fibroblast-stimulating activity, demonstrable by suitable dilutions of the supernates, coexisted with a number of other lymphokine activities such as lymphotoxin, proliferation inhibitory factor, and cloning inhibitory factor, which tend to reduce the numbers of function of fibroblasts. The increased content of collagen appeared to be the product of selected surviving and responding fibroblasts. The factor causing this increased collagen accumulation was nondialyzable and stable at -70 degrees C. It represents the first described lymphoid cell-derived activity capable of enhancing collagen accumulation. Fibroblast-stimulating activity may be implicated in the abnormal fibrosis seen in association with chronic inflammation in a variety of disease states. It may have special relevance to progressive systemic sclerosis.


An assessment of daily production and significance of thyroidal secretion of 3, 3', 5'-triiodothyronine (reverse T3) in man.

While 3, 3', 5'-triiodothyronine (reverse T3, rT3) has been detected both in human serum and in thyroglobulin, no quantitative assessment of its metabolic clearance rate (MCR), production rate (PR), or secretion by the thyroid is yet available. This study examines this information in euthyroid subjects and evaluates it in light of similar information about two other iodothyronines in the thyroid: 3, 5, 3'-triiodothyronine (T3) and thyroxine (T4). Thus, it was noted that rT3 is cleared from human serum at a much faster rate than are T3 and T4; the mean (+/-SE) MCR of rT3 was 76.7+/-5.4 liters/day in 10 subjects, whereas MCR-T3 and MCR-T4 in 8 of them were 26.0+/-2.2 liters/day and 1.02+/-0.06 liters/day, respectively. Therefore, even though the mean serum concentration of rT3, 48+/-2.8 ng/100 ml, was much lower than that (128+/-6.7 ng/100 ml) of T3, the mean PR-rT3 (36.5+/-2.8 mug/day) and the mean PR-T3 (33.5+/-3.7 mug/day) were similar; in comparison, the mean serum concentration and PR of T4 were 8.6+/-0.5 mug/100 ml and 87.0+/-3.9 mug/day, respecitvely. These data and those on the relative proportion of rT3, T3, and T4 in 10 thyroid glands were used to assess the significance of the contribution of thyroidal secretion to PR-rT3 and PR-T3. It was estimated that whereas thyroidal secretion may account for about 23.8% of serum T3 (or PR-T3), it may account for only about 2.5% of serum rT3 (or PR-rT3). Since peripheral metabolism of T4 is the only known source of rT3 and T3 other than the thyroidal secretion, it could be calculated that as much as 73.0 mug or 84% of daily PR-T4 may normally be metabolized by monodeiodination either to T3 or to rT3. MCR and PR of various iodothyronines were also examined in five cases with hepatic cirrhosis, where, as documented previously, serum rT3 may be elevated while serum T3 is diminished. The mean MCR-rT3 in these cases (41.0 liters/day) was clearly (P is less than 0.005) less than that (76.7 liters/day) in normal subjects. This was the case at a time when the mean MCR-T3 (26.7 liters/day) and the mean MCR-T4 (1.19 liters/day) did not differ from those (vide supra) in normal subjects. Distinct from changes in MCRs, the mean PR-rT3 (33.0 mug/day) was similar to, and the mean PR-T3 (10.1 mug/day) and the mean PR-T4 (66.4 mug/day) were much less than, the corresponding value in normal subjects. Furthermore, while the ratio of PR-rT3 and PR-T4 (rT3/T4) in individual patients was either supranormal or normal, the ratio of PR-T3 and PR-T4 (T3/T4) was clearly subnormal. The various data suggest that: (a) just as in the case of T3, the thyroid gland is a relatively minor source of rT3; peripheral metabolism of T4 is apparently its major source; (b) the bulk of T4 metabolized daily is monodeiodinated to T3 or to rT3; (c) monodeiodination may be an obligatory step in metabolism of T4; (d) monodeiodination of T4 to rT3 is maintained normal or is increased in hepatic cirrhosis at a time when monodeiodination of T4 to T3 is decreased.


Phosphaturic effect of dopamine in dogs. Possible role of intrarenally produced dopamine in phosphate regulation.

A possible role for dopamine in phosphate handling by the dog kidney was investigated by intrarenal artery infusions of dopamine. Dopamine increased fractional phosphate excretion both in the presence and absence of control of parathyroid hormone and calcitonin. In addition, dopamine increased both renal blood flow and sodium excretion, however, the phosphaturia was independent of these changes; since 30 min after completion of dopamine infusion, renal blood flow and sodium excretion returned to control levels and phosphate excretion remained elevated. For comparison, the vasodilator isoproterenol increased renal blood flow and sodium excretion without a significant change in fractional phosphate excretion. Thus, the phosphaturic effect of dopamine is probably independent of its vasodilator effect. The phosphaturic effect of dopamine could not be accounted for by subsequent conversion to norepinephrine, since norepinephrine was antiphosphaturic in the dog. The effect of endogenous dopamine on renal phosphate excretion was investigated by intrarenal infusion of the precursor dopa. Dopa was phosphaturic both in the presence and absence of parathyroid hormone and calcitonin. In dogs pretreated with carbidopa, which blocks conversion of dopa to dopamine, dopa was no longer phosphaturic, although the kidney remained responsive to dopamine. It is postulated that dopamine may play a role in the intrarenal regulation of phosphate excretion.


The effects of glucose and insulin on renal electrolyte transport.

The effects of hyperglycemia and hyperinsulinemia on renal handling of sodium, calcium, and phosphate were studied in dogs employing the recollection micropuncture technique. Subthreshold sustained hyperglycemia resulted in an isonatric inhibition of proximal tubular sodium, fluid, calcium, and phosphate reabsorption by 8-14%. Fractional excretion of sodium and phosphate, however, fell (P is less than 0.01) indicating that the increased delivery of these ions was reabsorbed in portions of the nephron distal to the site of puncture and in addition net sodium and phosphate transport was enhanced resulting in a significant antinatriuresis and antiphosphaturia. The creation of a steady state plateau of hyperinsulinemia while maintaining the blood glucose concentration of euglycemic levels mimicked the effects of hyperglycemia on proximal tubular transport and fractional excretion of sodium and calcium. Tubular fluid to plasma insulin ratio fell, similar to the hyperglycemic studies. These results suggest that the effects of hyperglycemia on renal handling of sodium and calcium may be mediated through changes in plasma insulin concentration. In contrast to hyperglycemia, however, hyperinsulinemia cuased a significant fall in tubular fluid to plasma phosphate ratio with enhanced proximal tubular phosphate reabsorption (P is less than 0.02). This occurred concomitantly with a significant inhibition of proximal tubular sodium transport. These data indicate that insulin has a direct effect on proximal tubular phosphate reabsorption, and this effect of insulin is masked by the presence of increased amounts of unreabsorbed glucose in the tubule that ensues when hyperinsulinemia occurs secondary to hyperglycemia. Fractional excretion of phosphate fell significantly during insulin infusion but unlike the hyperglycemic studies, the fall in phosphate excretion could be entirely accounted for by enhanced proximal reabsorption.


Intestinal myoelectric activity in response to live Vibrio cholerae and cholera enterotoxin.

The myoelectric response of the rabbit ileum was studied in response to live Vibrio cholerae culture, a whole cell lysate of cholera, and the purified enterotoxin. Each cholera preparation produced a series of highly organized migrating action potential complexes (MAPC). An MAPC was defined as action potential discharge with a duration of 2.5 s or longer, followed by similar activity on at least one other consecutive electrode site. The mean and modal onset time of MAPC activity occurred 4 h after the infection with live Vibrio cholerae culture, the freeze-dried whole cell lysate preparation, or the purified enterotoxin. After the onset of activity this pattern persisted for the duration of the recording period (up to 12 h). The MAPC had a mean propagation velocity of 0.85+/-0.07 cm/s (mean+/-SEM), which remained constant with time. Direct visual observation of the loop revealed that the MAPC's resulted in contractions that propelled intraluminal contents in an aborad direction. The mean fluid output from the 12-cm ileal loops was 6.4+/-1.1 ml/h (mean+/-SEM). Control experiments consisted of recordings from: (a) a ligated ileal loop into which nothing was placed; (b) a ligated ileal loop into which either uninfected culture broth or 0.9% NaCl solution was injected; (c) a ligated ileal loop infused with 0.9% NaCl solution at a rate of 11.2 ml/h, and (d) rapid injection of 1.0, 2.5, 5.0, or 10.0-ml boluses of 0.9% NaCl into the proximal catheter. MAPC activity was not observed in any of the control experiments. These studies indicate that in addition to a secretory component to cholera, there exists a highly organized MAPC that results in contractions that propel intraluminal contents in an aborad direction.


The mechanism whereby bile acid micelles increase the rate of fatty acid and cholesterol uptake into the intestinal mucosal cell.

Studies were undertaken to define the mechanism whereby bile acid facilitates fatty acid and cholesterol uptake into the intestinal mucosal cell. Initial studies showed that the rate of uptake (Jd) of several fatty acids and cholesterol was a linear function of the concentration of these molecules in the bulk phase if the concentration of bile acid was kept constant. In contrast, Jd decreased markedly when the concentration of bile acid was increased relative to that of the probe molecule but remained essentially constant when the concentration of both the bile acid and probe molecule was increased in parallel. In other studies Jd for lauric acid measured from solutions containing either 0 or 20 mM taurodeoxycholate and saturated with the fatty acid equaled 79.8+/-5.2 and 120.8+/-9.4 nmol.min(-1).100 mg(-1), respectively: after correction for unstirred layer resistance, however, the former value equaled 113.5+/-7.1 nmol.min(-1).100 mg(-1). Maximum values of Jd for the saturated fatty acids with 12, 16, and 18 carbons equaled 120.8+/-9.4, 24.1+/-3.2, and 13.6+/-1.1 nmol.min(-1).100 mg(-1), respectively. These values essentially equaled those derived by multiplying the maximum solubility times the passive permeability coefficients appropriate for each of these compounds. The theoretical equations were then derived that define the expected behavior of Jd for the various lipids under these different experimental circumstances where the mechanism of absorption was assumed to occur either by uptake of the whole micelle, during interaction of the micelle with an infinite number of sites on the microvillus membrane or through a monomer phase of lipid molecules in equilibrium with the micelle. The experimental results were consistent both qualitatively and quantitatively with the third model indicating that the principle role of the micelle in facilitating lipid absorption is to overcome unstirred layer resistance while the actual process of fatty acid and cholesterol absorption occurs through a monomer phase in equilibrium with the micelle.


Diagnostic potential of lysosomal hydrolases in body cavity effusions.

Hexosaminidase, alpha-mannosidase, beta-galactosidase, beta-glucuronidase, and arylsulphatase A were measured inperitoneal and pleural effusions from patients with benign, malignant, and inflammatory disorders. Compared with the benign transudates, all enzyme activities were moderately elevated in malignant effusions and markedly elevated in inflammatory effusions. The assay of hexosaminidase and and alpha-mannosidase indicated clearly the underlying pathology in most specimens studied. This method could be of clinical value when the cause of an effusion is in doubt, particularly since the diagnostic criteria are independent of the presence or absence of tumour cells in the aspirate.


A microplate enzyme-immunoassay for toxoplasma antibody.

A new test for the detection and measurement of toxoplasma antibody is described. Test sera are reacted with antigen-sensitized wells in micro-haemagglutination plates. Any attached antibody is shown by the addition of an enzyme-labelled antiglobulin followed by assay of the enzyme reaction with its substrate. The test is easy to carry out on a large scale, and there is a positive correlation between the results and dye test and haemagglutination test titres.


Susceptibility of urinary pathogens to various antimicrobial substances: a four-year study.

The results of testing all urinary pathogens from general practice and hospital practice for their sensitivity to a range of antimicrobial agents over the four years 1971-74 are presented. The changes observed in each situation with the passage of time are discussed. Resistance has become more frequent to those drugs to which resistance is commonly R factor-mediated. Less change has been observed in resistance to other antimicrobial agents.


Tuberculosis of the cervix: cytology as an aid to diagnosis.

Two cases of tuberculous cervicitis are presented. The presence in cervical smears of both epithelioid and Langhans' giant cells, which are described, should raise a strong suspicion of tuberculosis.


Introduction of a Vickers M300 analyser into the routine service of a hospital laboratory. 1. Installation, staffing, logistics.

A Vickers M300 analyser has been successfully installed in a busy hospital laboratory. A permanent team of operators is used, but this introduces problems for staff rotation, particularly for educational purposes. The vials used for the collection of samples are an essential feature of the system, but they present many difficulties and are no longer used for collecting samples from outpatients, or at remote sites. Direct costs during 1974, excluding staff and depreciation, amounted to approximately pounds 0-45 per patient sample: this is assessed on the rising workload during implementation. An increased workload could still be handled with existing staff. The siting of such high-capacity analysers constitutes an important policy decision.


Introduction of a Vickers M300 analyser into the routine service of a hospital laboratory. 2. Tests, methods, evaluation.

A Vickers M300 analyser has been introduced into a busy hospital laboratory; a profile of 13 biochemical tests is currently provided. Analytical performance has been assessed over one year of routine service operation, and mechanical performance has been assessed over two separate three-month periods. At its best, the M300 achieves high standards of precision and accuracy and has the advantage of unrivalled speed. Enzyme assay methods are less satisfactory than the kinetic assays in current use. Random analytical errors occur and are difficult to detect. The restricted facilities of the integral process-control computer are inadequate for effective quality control.


Conditions for collection of serum samples for the measurement of fibrin(ogen) degradation products by radioimmunoassay of fragment E.

A number of conditions have been assessed for the collection of serum samples for the measurement of fibrin(ogen) degradation products using a radioimmunoassay for degradation fragment E, which permits precise quantitation of differences. Blood should be collected using minimal venous occlusion into glass tubes containing 10 mg/ml of epsilon amino caproic acid and allowed to clot at 4 to 20 degrees C for at least 4 hours before centrifugation. The serum may be stored at 4 or -20 degrees C. Samples from patients receiving anticoagulant therapy should be treated with 10 IU of thrombin per ml.


Fibrin-fibrinogen degradation products in cerebrospinal fluid.

An increase in low molecular weight fibrin-fibrinogen degradation products (FDP) was demonstrated in cerebrospinal fluid (CSF) from 17 of 18 patients with bacterial or viral meningitis compared with 29 patients without meningitis. The CSF also showed an increase in coagulation proteins of molecular weight less than 90000 (factors VII, IX, and plasminogen) but did not contain fibrinogen (MW 340000) or plasminogen activator. It is concluded that low molecular weight FDP in the CSF in infective meningitis result from leakage through a damaged blood-CSF barrier rather than from local digestion of fibrin deposited on the meninges.


Systemic angioendotheliomatosis with metastases.

The clinical and pathological findings of a case of systemic angioendotheliomatosis are presented. The previously published cases of this disease are analysed. The present case is only the third to show clear evidence of metastasis.


Histometry of lymphoid infiltrate in the thyroid of primary thyrotoxicosis patients. Relation of extent of thyroiditis to preoperative drug treatment and postoperative hypothyroidism.

The thyroids of primary thyrotoxicosis patients prepared for partial thyroidectomy with propranolol contained much more lymphoid infiltrate than those prepared with carbimazole. No relation was found between the extent of lymphoid infiltrate in the thyroid and the development of postoperative hypothyroidism either between or within the two drug treatment groups. This study has shown that the extent of thyroid infiltrate should not be used as the major factor in predicting hypothyroidism after subtotal thyroidectomy for primary thyrotoxicosis.


Introduction of a rapid, simple radioimmunoassay and quality control scheme for thyroxine.

A simple radioimmunoassay has been developed for service purposes to determine serum total thyroxine levels. Only three additions are required, of standard or sample, labelled thyroxine and antibody in polyethylene glycol. After 2 hours' incubation at room temperature the antibody-bound and free fractions are separated by centrifugation. Serum total thyroxine levels were measured in 195 euthyroid subjects and it was established that normal values lay within the range 57 to 155 nmol/1. Serial blood samples taken over a 24-hour period, from 11 subjects, indicated that there was no circadian rhythm so that samples for total thyroxine assay can be taken at any time of the day. Similar results were obtained using serum or plasma. Satisfactory results were obtained for three quality control sera when measured by seven different laboratories using this method.


Biopterin derivatives in human body fluids and tissues.

Levels of biopterin derivatives in urine, serum, milk, cerebrospinal fluid, brain, and liver have been measured with the Crithidia fasciculata assay. Normal levels in serum and urine have been given and compared with those in a number of benign and malignant proliferative disorders, phenylketonuria, kidney disease, Parkinson's disease, schizophrenia, controlled epilepsy, rheumatoid arthritis, and pernicious anaemia. The active component of Crithidia factor in serum was 7,8-dihydrobiopterin. Tissue, urine, and some serum samples contained two active materials, the principal one being 7,8-dihydrobiopterin; a minor constituent was probably tetrahydrobiopterin. Serum biopterin levels following methotrexate administration were raised and subsequent administration of folic acid and 5-formyltetrahydrofolic acid further increased serum levels of biopterin derivatives; this was in contrast to the total absence of response to oral folates without prior methotrexate and to 5-methyltetrahydrofolic acid either with or without methotrexate being given.


The time of origin and the pattern of survival of neurons in the isthmo-optic nucleus of the chick.

The time of origin of the cells in the isthmo-optic nucleus (ION--the nucleus of origin of centrifugal fibers to the avian retina) has been determined in the chick by a variant of the cumulative labeling method, using 3H-thymidine autoradiography. All the neurons of the ION are generated (i.e., pass through their last phase of DNA synthesis) over a 50-hour period between the latter part of the fifth and the seventh days of incubation (stages 28--31 of the Hamburger and Hamilton ['51] series) but the cells come to be assembled within the nucleus along a distinct temporo-spatial gradient. The earliest-formed cells occupy the ventrolateral part of the nucleus while the last neurons to be generated come to lie along its dorsomedial margin. When the nucleus is numerically complete, around the eleventh day of incubation, it contains about 22,000 neurons, but between the thirteenth and seventeenth days (stages 39 through 43) this number is produced by nearly 60% to about 9,500 cells. Following the radical extirpation of the one optic cup or the circumscribed removal of the neural retina early on the third day of incubation, the cell loss in the contralateral ION is greatly accentuated, so that by the eighteenth day of incubation no cells remain in the nucleus. Serial counts of the numbers of cells in the nucleus on the side of the eye (or retinal) removals show that it, too, undergoes additional cell degeneration, so that by the end of the phase of naturally occurring neuronal loss, only about half the normal number of cells persist in these experimental animals.


Dendroaxonic synapses in the substantia gelatinosa glomeruli of the spinal trigeminal nucleus of the cat.

The glomeruli in the substantia gelatinosa layer of the spinal trigeminal nucleus of the cat contain three kinds of dendritic processes. One of these, the type 2 dendrite, contains large synaptic vesicles in its spine heads and in its shafts. The type 2 dendrite receivers axodendritic synapses from primary trigeminal afferent (C) axons and an occasional axodendritic synapse from small axonal (P) endings with small synaptic vesicles. The type 2 dendrites in turn form dendroaxonic synapses on the C endings. The dendroaxonic synapse and the axodendritic synapse of the C ending typically occur in reciprocal pairs. The axodendritic synapse usually lies in the depths of scalloped depressions in the surface of the C ending while the dendroaxonic synapse is found on the rim of the depression. Type 1 spines, i.e., dendritic spines receiving axodendritic synapses from the primary ending and lacking synaptic vesicles, also receive dendrodentritic synapses from type 2 dendrites. The type 2 dendrite with its large, rounded synaptic vesicles is considered to be excitatory at its dendroaxonic and dendrodendritic synapses. The type 2 dendrites course from glomerulus to glomerulus receiving their excitatory input through the axodendritic synapses of C axons. A type 2 dendrite, in response to C axon excitation would activate type 1 spines directly through their dendrodendritic synapses (C leads to 2 leads to 1) and indirectly by increasing transmitter release at the axodendritic synapses of the C axonal endings through their dendroaxonic synapses (2 leads to C leads to 1). The type 2 dendrites could serve two functions. First, they may prolong transmitter release from the axodendritic synapses of C axonal endings beyond the time of arrival of incoming potentials because of the reciprocal pairing of dendroaxonic and axodendritic synapses (C in equilibrium 2). Second, they may extend the spatial range of the excitatory output of active primary afferent axons to type 1 spines of glomeruli whose primary afferent axons may be inactive (C leads to 2 leads to 1).


The facial motor nucleus of the opossum: cytology and axosomatic synapses.

The normal cytology and synaptic organization of the opossum facial motor nucleus was determined by use of Nissl preparations, Golgi impregnations, one-micron plastic sections and electron microscopy. Neurons from all regions of the nucleus can be classified into three categories based on the size and shape of their perikarya, the distribution of Nissl substance and extent of their dendritic arbors. Large neurons (30--50 mu) exhibit numerous Nissl granules and their dendrites often span more than one-half the width of the nucleus in the transverse plane. Their proximal dendrites measure 4--7 mu in diameter, taper to 0.5--2 mu and display few spines. Medium neurons (20--30 mu) can be distinguished from the large nerve cells by their size and their less extensive dendritic arbor. Proximal dendrites measure 4--7 mu in diameter, tapes to 0.5--2 mu and are relativel aspinous. Small neurons (10--20 mu) display a thin rim of cytoplasm which is either uniformly basophilic or achromatic. The dendrites of these neurons are relatively unbranched and taper from 4--5 mu proximally to 0.5--2 mu distally. Their axons give off one or two collaterals within the nuclear borders. More than one-half the perikaryal membrane of large and medium neurons is covered by presynaptic profiles which were grouped into three primary categories based upon differences in their vesicle shape (spherical, pleomorphic, or ellipsoidal). Only a few axosomatic contacts are present on small neurons. Further differentiation of axosomatic synaptic endings is suggested by differences in vesicle size, in pre- and postsynaptic membrane densities and in the size of the synaptic terminals. Vesicle size was determined by a cybergraphic tablet and a PDP-12 computer system and is expressed as a mean area in nm2.


The facial motor nucleus of the opossum: synaptic endings on dendrites.

The diameters of dendrites of large, medium and small neurons (Falls and King, '76) were measured from Golgi impregnations of the opossum facial motor nucleus in order to classify dendritic profiles sectioned in the transverse plane in electron micrographs. Three categories of dendrites are described: (1) proximal (4-7 mu in diameter); (2) intermediate (2-4 mu in diameter) and (3) distal (0.5-2 mu in diameter). The distribution of axodendritic synaptic endings was determined, recognizing that the neuronal source of individual dendritic profiles when seen in the transverse plane of section cannot be absolutely determined in view of the overlap in size of the dendrites issuing from the three types of neurons. Presynaptic terminals were categorized according to vesicle shape (spherical, pleomorphic or ellipsoidal), vesicle size, terminal size, junctional characteristics and post synaptic distribution. The vesicle size is expressed as a mean area (nm2) and was determined by using a cybergraphic tablet and a PDP-12 computer system. In any given plane of section, synaptic terminals cover most of the membrane of proximal dendrites and decrease in number as intermediate and distal dendrites are encountered. In Golgi impregnations four classes of afferent fibers which ramify among the dendrites of facial neurons can be distinguished. As yet, their sources have not been identified. Possible sites of origin for presynaptic profiles are discussed in the context of previous light microscopic findings.


Effects of stimulation on embryonic activity in the chick.

Effects of stimulation on the stereotyped and coordinated movements in the chick embryo were investigated using the type of stimulation (regular loud clicks) that is known to accelerate development and the time of hatching. The number of coordinated movements, the amount and approximate size of all activity, heart rates and, in 20-day embryos, respiration rates were assessed during a 15-min control period, a 15-min period of stimulation, and a 15-min period of no stimulation. The results showed an increase in the number of coordinated movements, but not their size, and some changes in the heart and respiration rates after the onset of stimulation. These effects occurred in most cases only after several minutes of delay. It is suggested that this delay may be connected with the reciprocal relationship between the coordinated movements and random motility in that the former may be triggered more readily at times when the latter are suppressed.


Role of thyroid hormones in an abnormal grooming behavior in thyroidectomized cats and cats with pontile lesions.

Thyroidectomized cats and cats with pontile lesions exhibit the same abnormal grooming behavior. Investigations were conducted to determine whether the abnormal grooming behavior in cats with pontile lesions was attributable to a hypothyroid condition. Administration of thyroid hormones abolished the abnormal behavior of thyroidectomized cats but was without effect in cats with pontile lesions. Studies of brain and blood levels of thyroid hormones in cats with pontile lesions failed to detect any differences from normal cats, and normal effects of thyroid hormones on evoked potentials to light flashes and to clicks were obtained in cats with pontile lesions. It was concluded that any thyroid dysfunction that may exist in the cats with pontile lesions was not involved in the genesis of the abnormal grooming behavior. Because the abnormal grooming behavior fluctuates rhythmically during the year, an endocrine hypothesis was entertained as the source of the fluctuations; endocrine concomitants of the abnormal behavior were investigated with longitudinal studies. The urinary excretion of 11-hydroxycorticoids exhibited a significant negative correlation with the abnormal behavior. When compared with that of normal cats, the plasma level of 11-hydroxycorticoids of thyroidectomized cats was significantly decreased. Intramuscular injections of 11-hydroxycorticoids abolished the abnormal behavior in both thyroidectomized cats and cats with pontile lesions. These results considered together with previous findings indicate that thyroidectomy and pontile lesions induce similar changes in 11-hydroxycorticoid and serotonergic functions-changes that are critical features of the physiological bases of the abnormal grooming behavior in both groups.


Posttrial hippocampal, amygdaloid, and lateral hypothalamic electrical stimulation: effects on short- and long-term memory of an appetitive experience.

Water-deprived rats were given 150 licks of a 10% sucrose solution, followed 4 sec later by 30 sec of low-intensity electrical brain stimulation applied through bipolar electrodes implanted bilaterally in the dorsal hippocampus, amygdala, or lateral hypothalamus. Retention of the licking experience was then tested either 90 sec or 24 hr after the 150-lick experience by measuring the number of licks taken on an empty drinking tube. Rats that received hippocampal stimulation licked the empty tube less than controls did during the 24-hr, but not during the 90-sec, retention test. These data suggest that the hippocampus may be critically involved in processes (e.g., consolidation) mediating long-term, but not short-term, memory. Compared with controls, rats that received amygdala stimulation did not exhibit any changes in licking at either retention test. Rats that received lateral hypothalamic stimulation showed more licking compared with controls during the 90-sec, but not the 24-hr, retention test. Subsequent tests suggested that facilitation of short-term memory may have been influenced by stimulation-induced motivational changes.


Neutralization of sensory-input modification of seizure thresholds in rats.

In this series of experiments it was demonstrated that the result of exposing rats to various parameters of either intermittent sound or intermittent light stimulation significantly decreased the threshold to chemically induced generalized seizures. Furthermore, simultaneous presentation of intermittent light with intermittent sound resulted in a mutual intersensory inhibition (cancellation, neutralization) of the preexisting seizurogenic potential of each stimulus. In these cases, the seizure thresholds did not differ from control values. Several working hypotheses are extended to probe the mechanism of the above results. The probable significance of these observations are (a) that the corroboration of results of other experiments on bimodal inhibitory interactions in normal animals suggests that intersensory inhibition is a property of the nervous system presented simultaneously with certain inputs and (b) that if generalizable to humans, the principle of intersensory inhibition may find an application in the abortion of sensory-precipitated seizures.


Effects of serial unilateral and serial bilateral visual cortex lesions on brightness discrimination relearning in rats.

In Experiment 1, rats with serial unilateral (SU), serial bilateral (SB), or one-stage bilateral (B) visual cortex lesions (areas 17, 18 and 18a) were compared in their retention of a preoperatively learned brightness discrimination. Both the first-stage and second-stage lesions for the SU and SB groups involved the same amount of cortical tissue and the same composite areas of visual field projection in primary and extraprimary visual cortex. Groups SU and SB showed substantial savings of the brightness discrimination after the completed bilateral visual cortex removals, while Group B showed a complete postoperative loss. In Experiment 2, the effects of two additional types of serial bilateral visual cortex lesions were investigated. These involved the medial and then lateral aspects of visual cortex in two stages (ML), or vice versa (LM). Once again, the serial bilateral lesion groups (SB, ML, and LM) showed substantial savings of the discrimination after the completed lesion, while Group B showed a complete post-operative loss. These results demonstrate a general advantage of serial damage over one-stage damage to visual cortex in recovery of a preoperatively learned brightness discrimination and indicate that the serial lesion effect is not specific to interhemispheric relationships.


Chemical sympathectomy and two-way escape and avoidance learning in the rat.

Six experiments are reported on the effects of 2,4,5-trihydroxyphenylethyl-amine (6-hydroxydopamine) on two-way escape and avoidance learning. Rats were tested on either escape or avoidance learning at 80 days of age after chemical sympathectomy at birth or 40 or 80 days of age. Neonatal and chronic sympathectomy (at 40 days), but not acute sympathectomy (at 80 days), resulted in depressed escape learning. Avoidance learning was affected by neonatal sympathectomy and partially by acute sympathectomy. The results have implications for the role of the autonomic nervous system in escape-avoidance learning.


Topical cycloheximide in psoriasis.

Cycloheximide is an antimitotic and an inhibitor of the synthesis of both DNA and protein. Topical application in low concentrations (0.1%-0.2%) resulted in definite involution of psoriatic plaques. Because of its potency this experimental drug - not yet available for clinical use - was employed on small areas only and with extreme caution. Results of a full evaluation for safety, currently in progress, are being awaited with interest.


Ultrastructure of spindle cell squamous carcinoma.

A 63-year-old white male presented with a nine-month history of a nontender ulcerated lesion on his ear. Light microscopy demonstrated a moderately well circumscribed lesion in the dermis which abutted upon epidermis. There was no evidence of continuity between the tumor and overlying epithelium. The tumor was very cellular with an admixture of cells - spindle, polyhedral and bizarre giant cells. Mitotoc figures were abundant and frequently abnormal. We interpreted this lesion to have the clinical and pathologic features of an atypical fibroxanthoma (AFX). Ultrastructure, however, showed abundant tonofilaments and desmosomes indicative of an epithelial origin and therefore most consistent with a spindle cell squamous carcinoma (SCSC). It is urged that, when possible, electron microscopy be performed on problematic cases diagnosed either as an AFX or spindle cell squamous carcinoma since it is the most valid basis on which a correct diagnosis can be made.


Ultrastructural localization of acid phosphatase in normal and X-irradiated epidermis.

Acid phosphatase activity has been studied in normal and X-irradiated rat epidermis. A remarkable reaction increment was found in irradiated epithelium at the granular layer keeping the same pattern of distribution observed in the normal tissue. No reactive lysosomes or keratinosomes were observed in the cytochemical sections, while many keratinosomes but no lysosomes appeared in the electron microscopic preparations of irradiated animals. Irradiated epithelium is discussed as a suitable model to analyze the role of acid phosphatase in the keratinization mechanisms.


Ultrastructure of pustules in impetigo herpetiformis.

Typical pustular lesions of impetigo herpetiformis were examined by electron microscopy. In the upper layers of the epidermis degenerated keratinocytes and neutrophils lay close together and amorphous substances were seen within and outside the keratinocytes. Neutrophiles were not observed within the keratinocytes. The spongiform pustules of Kogoj observed with light microscopy were made up of a large number of neutrophils in a network of degenerated and stretched-out keratinocytes. Amorphous masses of high electron density which seemed to represent highly degenerated keratinocytes were observed among neutrophils.


Lobomycosis. An electronmicroscopic, histochemical and immunologic study.

A 73-year-old woman from Guyana had nodular keloidal lesions on her face which had persisted for 56 years. A biopsy of the lesion revealed lobomycosis, a rare fungal infection. Microscopically, numerous fungi were seen in giant cells and macrophages. The macrophages had abundant pink reticulated cytoplasm. These macrophages in lobomycosis, considered by some in the past to be granular myoblastoma cells, were rich in glycoproteins and appeared to be "Gaucher-like cells" in electronmicrographs. Further, a transition of macrophages containing organisms to Gaucher-like cells was noted. Antibodies to the fungus were demonstrated in the patient's serum by counter immunoelectrophoresis and indirect immunofluorescence studies. We postulate that the Gaucher-like cells formed because of the inability of host macrophages to digest glycoproteins in the capsule of this unusual fungus.


Amino terminal sequence and location of phosphate groups of the major human casein.

The amino acid sequence of the first 28 residues of the major human casein was determined. This protein in multiphosphorylated forms (0 to 5 phosphorous per molecule) was compared to cow beta-casein which is similar in composition but phosphorylated at a constant level. After sequencing the phosphate-free human casein, phosphorylated seryl and threonyl residues were located in three of the other phosphorylated forms by examining the aqueous layer of the phenylthiohydantoin conversion step during automatic liquid phase sequencing. Phosphate groups on specific seryl/threonyl residues suggest a biosynthetic mechanism involving stepwise phosphorylation or dephosphorylation.


Effect of alcohols on lactose solubility.

In tests of effects of high (70 to 90% volume/volume) and low (2 and 5% volume/volume) alcohol concentrations, solubility of lactose decreased with increased alcohol concentration and decreased as alcohol chain length increased. Since lactose has lower solubility in alcohol, crystallization would be expected to be speeded by increased supersaturation. Composition of precipitates formed by the action of ethanol changed with time; alpha-lactose precipitated more rapidly at first, then beta-lactose. Time of crystallization was related directly to the percentage of beta-lactose (therefore, the relation was inverse for total alpha). However, the percentage of alpha-hydrate increased with time and with water content. Agitation during crystallization increased production of beta-lactose. Composition of the lactose precipitate varied greatly with concentration of alcohol (ethanol). When ethanol concentration was low, only alpha-hydrate was precipitated whereas at higher concentrations stable anhydrous alpha lactose also was precipitated, with the percentage of total alpha decreasing while the percentage of beta-lactose increased. Crystal shape changed from prisms initially to partially or fully developed tomahawks as time went by or as the percentage of ethanol decreased, and crystal color increased with crystallization time and as ethanol percentage decreased. Choosing long-chain alcohols and controlling suitable parameters enabled recovery of greater amounts and more desirable forms of lactose.


Bovine milk intake and xanthine oxidase activity in blood serum.

Xanthine oxidase activity in blood serum was measured by a sensitive radio-enzymatic assay. Pigs receiving 7.6 liters of milk daily for 100 days did not show any detectable enzymatic activity in their blood Xanthine oxidase activity in blood serum of 25 human volunteers had an average of 6.7 milliunits per liter with a range of 0 to 34.6 milliunits per liter. Neither a causal nor statistically significant relationship existed between xanthine oxidase activity in blood and average daily milk consumption, age, or sex.


Influence of caloric density on energy intake by dairy cows.

Twenty-four Holstein cows were fed mixed diets of alfalfa hay and concentrate in a series of periods to determine the relationship between caloric density (mcal digestible energy/liter) of the diet and energy intake. Five diets with caloric densities of .58, .63, .68, .84, and 1.17 (as-fed form) were used. Steers were fed the diets in digestion trials to determine maintenance digestibility. Intake of digestible energy was adjusted for intake. Means of digestible energy intakes (kcal/day per kg. 75) were: 291.7, 324.0, 370.0, 371.4, and 369.1 for diets with caloric densities of .58, .63, .68, .68, .84, and 1.17. Physical fill was limiting for the two most dilute diets. The diet with caloric density of .68 represented the point above which physiological regulation was employed by the animals. The relationship between digestible energy intake (kcal/day per kg-75) body weight (Y) and caloric density (mcal/liter dry matter, as fed form) (X) was Y = 759.2X-148.4, simple correlation .99, for diets with caloric densities of .58, .63, and .68. Potential uses of this relationship in maximizing forage utilization and formulating diets to meet specific energy requirements are cited.


Factors influencing growth of rumen, liver, and other organs in kids weaned from milk replacers to solid foods.

From pooled data of many feeding trials with kids fed various types of solid foods from 37 days of age and slaughtered thereafter, relationships between such independent variables as age, cumulative intake of solid food, weight gain, and/or final body weight and such dependent variables as weights and such dependent variables as weights of stomach-compartments, heart, liver, kidneys, and spleen were analyzed. Growth of forestomachs, and especially rumen mucosa, relative to body weight was stimulated only in feeding periods of solid foods. Gain, intake of solid food, and final body weight were the best predictors for growth of forestomach-compartments, liver, and heart, respectively. Correlations were large between ruminoreticulum weight and liver weight and among weights of forestomach-compartments. Second degree polynomials were fitted to predict the curvilinear relationships between solid-food intake and either ruminoreticulum weight or rumen mucosal weight, and a linear equation was fitted to predict the relationship between solid-food intake and liver weight. Feeding of roughage increased rumen muscular and omasal weights. Growth of the rumen under stimulation of solid-food intake may be regulated by the same biological factors contributing to increased growth rate of the whole animal. In this situation, growth of the rumen may be correlated closely with growth of the liver.


Brucellosis status report.

Tremendous progress was made in the eradication of brucellosis for about 15 yr beginning in 1955. However, progress has been little in the last few years. This has been due to inflation and commitment of resources to other programs which had a higher priority at the time. We need additional resources, strengthened program procedures, and a renewed commitment on the part of all people and organizations involved in the eradication effort.


Epidermal surgery.

A review is given of the concepts, special tools, and techniques required for the intelligent practice of surgery on the epidermis and a new technique for removal of epidermal lesions by electrosurgical dissection is described.


Present status of surgical management of malignant melanoma.

Malignant melanoma has a distinctive appearance and has been clinically and histologically classified by Clark and Mihm. Using this classification a rationale for the surgical treatment of melanoma has been developed at the New York University Medical Center. The choice and extent of surgery is described. Early detection of melanoma and prompt surgical attention can significantly reduce the mortality from this neoplasm.


Wedge resection of the pinna.

In the treatment of large carcinomas whose local removal would be mutilating to a greater or lesser extent, wedge resection of the ear can preserve the normal appearance and function of the pinna. No significant complications have occurred in any of the patients so treated.


Basal-cell epithelioma occurring in a smallpox vaccination scar.

A patient with basal-cell epithelioma occurring in a smallpox vaccination site is reported. The association is probably not a chance occurrence.


Surgical treatment of hidradenitis suppurativa.

Hidradenitis suppurativa is a chronic, recurrent, suppurative inflammation of apocrine glands. Morbidity is high and serious complications may occur. A technique of surgical treatment utilizing an electrosurgical unit to provide cutting current is described. This allows rapid unroofing and cauterization of extensive cysts and sinus tracts.


Squamous-cell and basal-cell carcinomas in black patients.

Squamous-cell and basal-cell carcinomas of the skin are rare in black patients. Etiologic factors, body distribution, and aggressiveness of these cancers in blacks are discussed. The roles of ultraviolet light and pigment dispersion are noted.


Tumor on shoulder arising after excision of a cyst.

A primary cutaneous myxosarcoma of the skin developed at the site from which an epithelial cyst had been excised. Following this surgical excision a "keloid" developed which was injected with intralesional corticosteroids. At this site an enlarging tumor developed which was initially thought to be myxoma on the histologic examination but subsequently gave rise to a solitary lymphnode metastasis. Wide local excision of the primary lesion with skin graft and radical axillary lymphadenectomy resulted in cure. Now, nine years post surgery the only problem that remains is repeated bouts of acute cellulitis in the slightly lymphe dematous upper extremity. Myxosarcomas of the skin are rare tumours. Metastases from such lesions are exceedingly rare.


Histopathologic considerations in the management of skin cancer.

A knowledge of the pathology and biologic behavior of skin cancers is essential for accurate diagnosis and effective treatment. The dual nature and newer concepts of squamous-cell carcinoma and basal-cell epithelioma are reviewed. Some difficulties encountered in differentiating the two are discussed, including the "basosquamous" problem, squamous metaplasia, and pseudoepitheliomatous hyperplasia. Sclerosis within basal-cell epitheliomas is discussed, specifically relating to its importance in treatment. Follicular involvement in epidermal malignant and pre-malignant lesions is presented as a possible cause of recurrence.


Excisional electrosurgery (endothermy) in dermatology.

The physical characteristics of electrosurgery are reviewed and the peculiar behavior of the cutting current is described. The indications for excisional by electrosurgery are discussed and the procedure is described. The advantages and disadvantages of the method are given. It is concluded that the overall results of excisional electrosurgery compare very favorably with other surgical procedures on the skin.


Little tips with big rewards.

We all would like to reduce the pain of injecting a local anesthetic, to simplify the treatment of xanthelasma palpebrarum and chondrodermatitis nodularis; and to improve methods of shave excision of nevi and cosmetic extirpation of large seborrheic keratoses. Here are some tips that might help.


Diseases of the oral mucosa.

Lesions of the oral mucosa are numerous. The purpose of this article is to review the more common and most serious of them, to explore some basic-science aspects of their development and to discuss prevention and treatment.


Dermabrasion for showers of seborrheic keratoses.

Seborrheic keratoses, skin tumors of undetermined etiology, are easily traumatized with consequent local irritation and inflammation, and are cosmetically undesirable. They can occasionally undergo malignant degeneration. A rapid increase in size and number of these lesions is thought by many to be a sign of internal malignancy. Dermabrasion using a diamond fraise is a simple and fast method for removing multiple seborrheic keratoses with minimal trauma, rapid healing, and good cosmetic results.


Precipitating antibodies in a midwest dairy farming population toward the antigens associated with farmer's lung disease.

A survey of the frequency of precipitins to the antigens of the thermophilic actinomycetes and Aspergillus species was conducted on serum samples from 1,045 farmers obtained at a 3-day exposition on modern farm equipment and farming practices in central Wisconsin. Each farmer filled out a questionnaire including socioeconomic information, lung disease history, exposure history, and smoking history. Precipitins were detected by the double-diffusion method. The antigen panel included eight thermophilic actinomycetes species and a mixture of Aspergillus species. Precipitins were found in the sera of 93 famers (8.9%). The distribution of positive precipitins was: Micropolyspora faeni 63 (67.7%), Thermoactinomyces vulgaris 7 (7.5%), Thermomonospora viridis 2 (2.2%), M. faeni + T. viridis 16 (17.2%), M. faeni + T. vulgaris 1 (1.1%), Aspergillus species 4 (4.3%). Of all the parameters tested for in the questionnaire, those with positive serology differed significantly from the whole population only in that a higher proportion of the positives reported exposure to silo gas and illness after uncapping silos. Comparison of the size of the farm and the number of dairy cows in the state of Wisconsin with the population samples indicated that the sample population was skewed toward those with larger farms and larger dairy herds. This study confirms that a significant proportion of the farm population in Wisconsin does have precipitins to the microorganisms associated with farmer's lung disease. Follow-up studies to establish the relationship between the positive precipitin reactions to the presence of clinical disease are now under way.


Immunological studies of the effect of whole body insect extracts in the treatment of stinging insect allergy.

Specific IgE antibodies and total antibodies reacting with bee venom and yellow jacket venom were measured in sequential serum samples of insect-sensitive individuals. Venom-specific IgE decreased as a function of time and was not significantly affected by treatment with whole body extracts. There was no stimulation of total antibodies reacting with bee venom phospholipase A2 (PLA) following treatment with whole bee body extracts. These studies suggest that as measured by these parameters, whole body insect extracts used in the usual recommended doses are immunologically ineffective antigens.


Metabolic changes in exercise-induced and methacholine-induced bronchoconstriction.

Metabolic and physiological responses to graded exercise and methacholine challenge were investigated in asthmatics with or without exercise-induced bronchoconstriction. The results showed that after methacholine challenge, free fatty acid levels increased only in patients with exercise-induced asthma, while they increased in both groups of asthmatics after treadmill exercise. No significant changes were noted in plasma lactic acid levels, ventilation, or oxygen consumption among the groups studied. These data suggest that asthmatics with exercise-induced bronchoconstriction may differ from other asthmatics in some of their metabolic responses.


Immunologically induced lung disease in guinea pigs. A comparison of ovalbumin and pigeon serum as antigens.

This study was conducted to compare the capacity of pigeon serum (PS), an antigen (Ag) associated with hypersensitivity pneumonitis (HP), and ovalbumin (OA) in the induction of immunologic lung disease in guinea pigs (gp). Whereas OA was very effective in inducing a severe pneumonitis, PS failed to produce significant disease. A determination of the antibody (Ab) responses in OA- or PS-sensitized GP revealed that total Ab activity, as well as specific IgG1, and IgG2 responses, were not significantly different in the two groups. There was, however, a markedly higher IgE-like Ab response to OA than to PS. Thus, there was a striking correlation between specific IgE synthesis and the production of immunologic lung disease. The disease resembled immune complex disease histologically, and we suggest that the IgE antibody may function as a preceding "anaphylactic trigger" mechanism for the lodging of complement-fixing Ag-Ab complexes in the vasculature of the lung. It is further suggested that PS may be a poor Ag for the induction of IgE synthesis in guinea pigs.


Pollen season severity and meteorologic parameters in central New Jersey.

A systematic search is made for optimum parameters to be used in forecasting pollen season severity. All possible averaging periods for daily maximum temperature, minimum temperature, average temperature, and precipitation are tested. Useful positive correlations are found with spring and summer daily minimum temperatures and average temperatures, and with midspring precipitation amounts.


Energy expenditure and consumption of mature, pregnant and lactating women.

Activity patterns, energy expenditure, and energy and protein consumption of mature women were determined during preganncy and lactation. Homemakers of average economic status from a mixed population were not significantly more active than teenage women, but the range of activities was greater. Average energy output for the latter half of gestation was 2,200 to 2,300 kcal per day; per unit of body weight, the mean was 32.5 +/- 4 kcal per kilogram for the twenty-week period. A small decline of 6 per cent in energy expenditure was noted near term. Allowing for deposition of fetal and material tissue, the average metabolizable energy need for this group was about 35 to 36 kcal per kilogram for the latter half of pregnancy. These data show that a pregnant woman of reference body weight (68 kg.) may vary in energy output by 800 to 900 kcal per day, depending on occupation. Homemakers with small children and especially those who work outside the home constitute a high energy work category. Thus, the need for considering work pace and work load, as well as body mass, in estimating the energy requirement during pregnancy was confirmed. Average daily energy intake reported was 1,955 kcal or 28.5 kcal per kilogram for the latter half of gestation. A mean protein intake of 1.17 gm. per kilogram per day represented 17 per cent of gross energy consumed. It is questionable whether the energy level consumed by these women was sufficient to maintain positive nitrogen balance on the days recorded. Lactating homemakers expended an average of 30 kcal per kilogram per day, exclusive of milk production. Energy intake was 30 kcal per kilogram, and was equal to 74 per cent of need when adjusted for milk production. Non-lactating women expended 34 kcal per kilogram per day, 13 per cent above the values for lactating women. Average energy intake of non-lactating women was 19 kcal per kilogram, with protein intake representing 19 per cent of energy consumed for both groups.


The Comprehensive Nutrition Action Program in Arizona.

Through the Comprehensive Nutrition Action Program for Arizona (funded by the Nutrition Program, Center for Disease Control, Atlanta), a nutrition care delivery system was developed. Four sub-systems-screening, referral, monitoring, and the development of community nutrition workers--allowed for the identification of problems, intervention, utilization of other agencies, and expansion of manpower with community nutrition/health workers. Improvement in nutritional status was demonstrated by this program. As a result, it has been expanded.


Microcrystalline cellulose replacement in cakes and biscuits.

Sensory and physical characteristics of cakes and biscuits with up to 60 per cent of flour replaced, volume for volume, with microcrystalline cellulose were compared with those of the respective controls. High quality products were obtained at replacement levels of 40 and 20 per cent, respectively, in lean-formula cakes and in biscuits. At these replacement levels, caloric density was reduced by 12 and 10 per cent, respectively, in the cakes and biscuits.


Noninvasive differentiation of cardiomyopathy from coronary disease; pulse transmission time and systolic time intervals.

Systolic time intervals and central pulse transmission time were compared in cardiomyopathy (CM) patients and age-and sex-matched patients with coronary artery disease (CAD). Significant differences were found in pulse transmission time (shorter in the CAD patients (p less than .001) and in the pre-ejection period (p less than .001), ejection time index (p less than .05), and in the ratio of the pre-ejection period to the left ventricular ejection time (p less than .001); all were markedly abnormal in the CM group. Pulse transmission time differences may have been the result of more generalized arteriosclerotic disease in the CAD patients. It appears that both systolic time intervals and pulse transmission time clearly distinguish CM from CAD.


Free radical theory of aging: effect of dietary fat on central nervous system function.

Free radical reactions have been implicated in aging. A rise in the level of random free radical reactions in a biologic system might have a greater effect on the central nervous system (CNS) than elsewhere, partly because of the presence of glial cells and the unique connections between neurons. To evaluate this possibility, some animal experiments were conducted. The initial experiment involved old male Sprague-Dawley rats fed (since shortly after weaning) with semisynthetic diets characterized by fat differing in amount or degree of unsaturation. The number of errors made in a Hebb-Williams maze was determined and found to be higher as the amount or degree of unsaturation of the fat was increased. Likewise rats aged 6 and 9 months, fed semisynthetic diets containing 20 percent by weight of lard, oleinate, or safflower oil +alpha-tocopherol performed significantly better in a discrimination learning situation (Skinner box) than did rats fed a diet containing 20 percent by weight of safflower oil. The diets employed in these studies did not have a significant effect on the mortality rates. These results are compatible with the possibility that enhancing the level of lipid peroxidation has an adverse effect on the CNS, out of proportion to the effect on the body as a whole, as measured by the mortality rate.


Sleep deprivation in the treatment of elderly depressed patients.

Fifteen elderly depressed patients were treated by 36-hour sleep deprivation (SD). The depression was unipolar in 3 cases, bipolar in 3, and secondary in 4. Nine of the 15 patients responded to SD, and 6 had a remission (1 with SD alone and 5 with SD plus an antidepressant drug). Some of the remaining 6 patients might have responded if the treatment had not been interrupted for various reasons. These favorable results in elderly patients were better than anticipated. SD was well tolerated, although in one patient with bipolar depression a manic attack was precipitated. The effectiveness of SD poses interesting theoretic questions.


Internal carotid arterial occlusion simulating extracerebral hematoma: relation to diagnosis of stroke.

In 2 elderly male patients, a progressive neurologic deficit and alteration of consciousness developed following substantial head trauma succeeded by an asymptomatic "lucid" interval. The sequence of events and the associated electroencephalographic, echoencephalographic and cerebrospinal fluid findings strongly suggested the presence of an extracerebral hematoma. The cerebral angiographic study, however, disclosed complete occlusion of the extracranial portion of the internal carotid artery in both patients. This report stresses the diagnostic difficulties sometimes encountered in relation to stroke patients.


The problem of pressure sores in a nursing home population: statistical data.

This study on 93 patients was conducted at a skilled nursing facility. Twenty-two of the 93 patients had pressure sores; 34 of the 54 sores were present when the patients were admitted and 20 developed after admission. Only 5 of the 54 sores healed. Healing times ranged from 15 to 55 days after the beginning of treatment. Thirteen deaths occurred among the 71 patients without sores, and 17 deaths among the 22 patients with sores. Specific treatment regimens varied (cleaning solutions, antimicrobial drugs, heat, enzymatic debridement) but always included frequent changes in the position of the patient. Ten of the 22 patients with sores were anemic vs. 19 of the 71 patients without sores. Half of the patients with sores were maintained with tube feedings whereas more than half of the patients without sores received regular house diets. Pressure sores are a serious problem among geriatric patients in nursing homes. All those who care for the aged should be fully aware of the procedures involved in the prevention and management of these sores.


Community intervention with the elderly: a social network approach.

A technique is described for using the advantages of a social systems approach when working with elderly persons in psychiatric distress. The technique is based on the assumption that the solution to a variety of human predicaments lies within the collective instrumental and affective resources of the client's social network. The vehicle for accomplishing this objective is the "Network Session" during which a mental health professional meets with the elderly person and members of his/her social network to help resolve the difficulty. A case report demonstrating use of the technique is included.


Effect of prolonged bran administration on serum levels of cholesterol, ionized calcium and iron in the elderly.

A significant lowering of serum cholesterol, ionized calcium and iron levels was demonstrated in 27 elderly patients given 10 or 20 gm of unprocessed wheat bran daily in the diet for six weeks.


Radiobiochemistry of phytodrugs: I. role of juvenile hormones and analogs in the biosynthesis of proteins and RNA in drosophila larvae.

The present communication concerns the investigations of the effect of Juvenile Hormone (J.H.) and its analogs (J.H.A.), such as Altosid (R) and Altozar(R), used as insecticides, on the biosynthesis of proteins and RNA in subcellular preparations of drosophila larvae with Leucine-U-14C and Uridine-2-14C respectively. The incorporation of leucine into proteins decreases progressively from crude homogenate to the microsome-free fraction in the same way as the degradation of the compounds. During RNA biosynthesis the incorporation of uridine increases from the crude homogenate up to mithocondria-free fraction and with Altosid is always below the test controls. The reduced incorporation of uridine into RNA and leucine into proteins in the presence of Altosid and Altozar indicates a genetic action of this compounds comparable to that of natural J.H. but slightly more efficient.


The evaluation of a radio gas-chromatographic system for the detection of trace amounts of labelled insecticides.

A radio gas-chromatographic system consisting of a gas chromatograph, combustion furnace, proportional counter and guard scintillation counter has been studied for the analysis of trace amounts of 14C-labelled organophosphorus insecticides. The proportional detector is a concentration sensitive detector requiring strict flow control. Sensitivity depends on the concentration of the quenching gas used. Using 14C-ethyl parathion and 14C-metamidophos as model compounds a linear range of 3500-5000 was measured. A sensitivity of 0.98 cpm/pCi and a detection limit of 24pCi were obtained which allows the detection of 0.33 ng if the specific activity of the insecticide is 21.5 mCi/mMol. The system must be decontaminated in order to achieve optimum detection levels by passage of oxygen through the combustion tube and by periodically injecting the corresponding non-labelled compound into the column. A solvent effect was found to interfere in the analysis due to the sudden production and expansion of CO2. This effect was eliminated by choosing column conditions to prevent elution near the solvent.  The system was tested by analyzing a mixture of 37 ppb and 23 ppb of ethyl parathion and its oxygen analog in orange juice.


Studies on the terminal stages of antibody-complement mediated killing of a tumor cell. III. Effect of membrane active agents.

A comparison of the events leading from T to dead cell and E to E ghost and hemoglobin was made. Transformation of T to dead cells was inhibitable by EDTA, NaF, histamine, DMSO, cytochalasin B and 3'5' cAMP. Transformation of E to E ghost and hemoglobin was only inhibited by EDTA.


Inflammatory process in murine lymphocytic choriomeningitis is maximal in H-2K or H-2D compatible interactions.

Capacity to transfer adoptively fatal lymphocytic choriomeningitis (LCM) to immunosuppressed, virus-infected recipients is a property of H-2 compatible, non-Ig-bearing virus-immune lymphocytes. Severe meningitis is recognized when donor and recipient share at least one allele at either H-2K or H-2D. Presence of unshared H-2 genes is not obviously inhibitory, and identity at the immune response (Ir) region of the H-2 gene complex is neither sufficient nor necessary. The same constraint applies to cytotoxic T cell activity in vitro; lymphocytes and virus-infected targets must be compatible for a minimum of one allele mapping at H-2K or H-2D. The present findings thus support the concept that populations of T cells, which are cytotoxic in vitro, also mediate inflammatory process in vivo and are a major, if not the only, effector population in murine LCM.


Antibody-mediated basophil accumulations in cutaneous hypersensitivity reactions of guinea pigs.

Cutaneous basophil hypersensitivity (CBH) was studied in guinea pigs by using simplified histologic techniques. Animals immunized with oxazolone or picryl conjugates of keyhole limpet hemocyanin (KLH) emulsified with complete (CFA) or incomplete Freund's adjunvant (IFA) were found to have hapten-specific cutaneous basophil reactions when skin tested at 1 week with oxazolone or picryl chloride contant painting or intradermal injection of oxazolone or picryl-conjugated human serum albumin, respectively. Thus, hapten-specific cutaneous basophil reactions were present in guinea pigs immunized with CFA for classical delayed hypersensitivity, and in animals immunized with IFA for Jones-Mote reactions. Hapten-specific 24-hr cutaneous basophil reactions were passively transferred with immune serum from donors sensitized with conjugates of oxazolone or picryl-KLH in CFA or IFA, and with serum from oxazolone contact-sensitized animals as well. As little as 0.5 ml sera obtained from donors 1 week after immunization could systemically transfer cutaneous basophil reactions. It is likely that hapten-specific cutaneous basophil reactions are mediated by small quantities of serum antibodies. We conclude that antibody-mediated cutaneous basophil reactions may be distinctive hypersensitivity responses that can be distinguished from classical anaphylactic, Arthus, and delayed hypersensitivities. It is suggested that CBH reactions are heterogeneous and that antibody products of B lymphocytes, and factors probably derived from T lymphocytes, play a role in basophil accumulations at cutaneous hypersensitivity reactions.


Immunosuppression of experimental autoimmune myasthenia gravis by hydrocortisone and azathioprine.

Experimental autoimmune myasthenia gravis (EAMG), induced in rabbits by injection of acetylcholine receptor (AChR) from Torpedo californica, was suppressed by appropriate treatment with hydrocortisone or with azathioprine. Administration of hydrocortisone in gradually increasing doses, starting at the time of immunization with the receptor, prevented exacerbation of the disease in the early stages of treatment, as was the case when hydrocortisone was administered in high doses from the beginning. Prolonged administration of the antimetabolite azathioprine (Imuran) prevented the appearance of EAMG, for at least 4 months, in rabbits immunized with AChR. Cell-mediated immunity to AChR was demonstrated to be significantly decreased in such treated animals. The effects of hydrocortisone and azathioprine on EAMG support the view that the disease involves an immunologically cell-mediated mechanism and indicate that the experimental disease can serve as a useful model for chemotherapy of the human disease.


Eosinophil chemotactic factor (ECF). II. Release from human polymorphonuclear leukocytes during phagocytosis.

Phagocytosis was found to stimulate ECF release from neutrophils (PMN). Human PMN (less than 98%) were exposed to zymosan (Z), zymosan-coated with complement (Z(x)), or zymosan in the presence of serum (Z(s)). The release of ECF was shown to be time- and dose-dependent. Like ionophore-induced ECF, phagocytosis-derived ECF preferentially attracted and deactivated eosinophils. Chromatography on Sephadex G-25 revealed an elution pattern similar to the antigen-induced, basophil-derived and to the ionophore-induced PMN-derived ECF. The addition of complement either as Z(x) or Z(s) accelerated the release of ECF. With both stimuli, the initial kinetics of ECF paralleled the release of beta-glucuronidase and NBT reduction. With Z alone, beta-glucuronidase release and NBT reduction were negligible, whereas the amount of ECF released was similar to that induced by Z(x). In the presence of serum Z(s), decreased activity was noted. Supernatants of phagocytosis at late time periods showed less activity than early supernatants, suggesting that ECF might be inactivated. These data indicate that phagocytosis causes the release of an ECF, which appears to be similar to the IgE-induced ECF-A.


Chemotactic responses of normal human basophils to C5a and to lymphocyte-derived chemotactic factor.

Peripheral blood basophils from normal human subjects responded chemotactically to the complement component C5a and also to the supernatant fluid from cultured human lymphocytes stimulated by mitogens or specific antigen. Basophil chemotactic factor (BCF) was detected in stimulated lymphocyte cultures within 24 hr and increased for the first 3 days. BCF was found after stimulation of lymphocytes by B or T cell mitogens as well as by specific antigen. The amount of BCF in culture fluids was independent of cell proliferation. Monocyte chemotactic factor (MCF) and BCF could not be separated from one another by Sephadex or DEAE chromatography. Both activities were eluted from Sephadex G-100 columns in a m.w. region of about 15,000 daltons and were found after ion exchange chromatography on DEAE in a peak eluted by 0.1 M NaCl. These findings suggest that mitogen or antigen-stimulated lymphocytes secrete a molecule that is chemotactic for both monocytes and basophils. Thus, in the absence of a basophil-specific chemotactic molecule, the preferential accumulation of basophils in certain types of delayed hypersensitivity skin reactions must be due to additional factors such as basophil chemotaxis augmentation factor.


Separation of populations of sensitized lymphoid cells into fractions inhibiting and fractions enhancing syngeneic tumor growth in vivo.

Separation of subpopulations of lymphoid cells sensitized against a syngeneic tumor was attempted by means of velocity sedimentation and the fractions were injected together with tumor cells into syngeneic hosts. Spleen cells from tumor-bearing mice or spleen cells sensitized on tumor cell monolayers could be fractionated into a subpopulation capable of inhibiting growth of the same tumor and a separable fraction which enhanced tumor growth. Activity of the tumor-inhibiting lymphocytes was not apparent in the presence of the tumor-enhancing cells. The former activity was associated with small lymphocytes and the latter with injection of larger cells. Preliminary experiments investigated the developmental relation between the cells responsible for these opposing activities.


Studies on glycopeptide released by trypsin from sheep erythrocytes.

Pretreatment of sheep erythrocytes with trypsin abolishes their specific binding and rosette formation with human T lymphocytes. A glycopeptide containing sialic acid is released from the intact sheep erythrocytes by incubation with trypsin and purified. This glycopeptide contains activity that can be bound to T lymphocytes and produces inhibition of rosette formation. This component with a m.w. of about 10,000 contains galactose, acetylglucosamine, acetylgalactosamine, sialic acid, and serine. These results suggest that the glycopeptide released by trypsin treatment may contain the site of the T cell receptor of sheep erythrocytes.


Interaction of soluble C3 fragments with guinea pig lymphocytes. Comparison of effects of C3a, C3b, C3c, and C3d on lymphokine production and lymphocyte proliferation.

C3a, C3b, C3c, and C3d were generated from purified guinea pig C3 by trypsin treatment. These fragments were characterized immunochemically and functionally by rosette inhibition. C3b is capable of binding to both C3b and C3d receptors on lymphocytes whereas C3d binds only to C3D receptors. C3b stimulates guinea pig spleen cells to elaborate a macrophage chemotactic factor which is similar in m.w. to that generated in response to PHA or LPS and is antigenically unrelated to C3 or C5. In contrast, neither C3a, C3c, or C3d stimulate guinea pig lymphocytes. Neither C3 nor any of its major fragments induce cellular proliferation. These data are compatible with the hypothesis that C3b triggers spleen cells to release a macrophage chemotactic factor by cross-linking C3b and C3d receptors.