Regulation of plasma lecithin:cholesterol acyltransferase in man. I. Increased activity in primary hypertriglyceridemia.

Patients with primary hypertriglyceridemia have been reported to manifest increased in vivo turnover of plasma cholesteryl esters. To ascertain if this is due to plasma lecithin:cholesterol acyltransferase (LCAT) and to explore a possible link between triglyceride and cholesteryl ester turnover, we have measured LCAT in 15 patients with Type IV, 2 with Type V, 1 with Type III, and 9 with Type II B hyperlipoproteinemia. LCAT was significantly elevated (p less than 0.001) in hypertriglyceridemic subjects, regardless of lipoprotein pattern. In the Type IV group, but not in normal subjects, LCAT correlated significantly with measures of very low-density lipoprotein (VLDL) elevation, including plasma triglycerides and particularly VLDL-unesterified cholesterol, but not with body weight or substrate high-density lipoprotein (HDL) lipid levels. On repeated determinations in individual subjects, a relationship between triglyceride fluctuations and LCAT could be demonstrated in only one subject over an extreme range of triglyceride levels. Analysis of lipoprotein lipids revealed that the ester:free cholesterol ratio in VLDL was increased in hypertriglyceridemia, but was not correlated with enzyme level. In vitro removal of endogenous VLDL or addition of VLDL from lipemic plasmas to normal plasmas was without effect on enzyme activity. Regulation of enzyme activity does not appear to be a direct function of VLDL level.


Catabolism of canine apolipoprotein A-I: purification, catabolic rate, organs of catabolism, and the liver subcellular catabolic site.

The catabolic site(s) of plasma lipoproteins and apolipoproteins has not been established. We have purified one of the major apolipoproteins and explored its catabolic rate and its site of catabolism. Apolipoprotein (Apo) A-1 was purified by column chromatography of canine high-density lipoprotein (HDL) subfraction, HDL3, which was totally delipidated; the purified Apo A-1, molecular weight 28,000, had only one precipitin line to either anti-Apo A-1, or anti-Apo HDL3 serum. Apo A-I was then iodinated, its catabolic rate measured, and the various organs and the subcellular sites of liver involved in the catabolism were investigated. The T 1/2 of 125I-Apo A-I in HDL3 was 3.33 +/- 0.08 days and in plasma 3.52 +/- 0.17 days. The disappearance curve of radioactivity recovered in density fractions d less than 1.110 and d greater than 1.210 Gm. per milliliter was the same as that in HDL3, suggesting identical Apo A-I catabolism in each density class. The liver and kidney absorbed more radioactivity than did the spleen, heart, lung, and intestine. In subcellular fractions of liver, radioactivity was predominantly recovered in the light mitochondrial fraction. There was a statistically significant correlation between the acid phosphatase relative specific activity and the relative specific radioactivity in each subcellular fraction examined at several time intervals after the injection of 125I-Apo A-I. Direct organelle isolation and demonstration of labeled apoplipoprotein uptake indicate that liver lysosomes may play an important role in the catabolism of Apo A-I, as may those of the kidney.


Benign mixed tumour of heterotopic salivary gland tissue in upper neck. Report of a case with a review of the literature on heterotopic salivary gland tissue.

A case of benign mixed tumour arising in ectopic salivary gland tissue of the mucous type is described. The tumour was located at the angle of the right mandible, in the upper middle third of the neck, along the anterior border of the sternomastoid muscle. This observation is substantiated by a pathomorphological study. A comprehensive review of the world literature on heteroptic salivary gland tissue has also been carried out, together with a search of the possible pathogenesis which seems to be connected with an anomalous embryonic development within the branchial apparatus.


Measurement of two plasma triglyceride lipases by an immunochemical method: studies in patients with hypertriglyceridemia.

Postheparin plasma lipolytic activity consists of two hydrolytic activities, hepatic triglyceride lipase and lipoprotein lipase. These two enzymes were separated and partially purified by means of ammonium sulfate precipitation and affinity chromatography using Sepharose with covalently linked heparin and concanavalin A, respectively. Antibodies were produced against hepatic triglyceride lipase and they did not cross react with lipoprotein lipase. Optimal conditions for selective precipitation of hepatic lipase and specific measurement of these two lipases were investigated. This method was applied to the study of 15 patients with hypertriglyceridemia and 8 patients with familial lecithin-cholesterol-acyltransferase deficiency of whom 6 also had a marked elevated plasma triglyceride concentration. All patients had normal values of hepatic plasma lipase. All 8 patients with Type I and 2 of 4 patients with Type V hyperlipoproteinemia had lipoprotein lipase activities that were markedly reduced. The patients with Type III hyperlipoproteinemia and all 8 patients with lecithin-cholesterol-acyltransferase deficiency also had normal lipoprotein lipase values. These studies emphasize the necessity for differentiating between triglyceride lipase activity of hepatic and extrahepatic origin in evaluating patients with impaired triglyceride metabolism.


Mechanism of salt-mediated inhibition of lipoprotein lipase.

The activity of lipoprotein lipase isolated from rat postheparin plasma has been determined with synthetic lipids, in the presence and absence of apoprotein of the natural substrate very low density lipoprotein, as a function of medium ion-pair concentration of a number of different inorganic salts. The several kinetic effects of lipoprotein protein on lipase activity were specifically and quantitatively reversed in the presence of molar sodium chloride or solutions of equivalent effective ion concentrations of other salts. Salt-mediated inhibition was fully reversible by silution and was independent of substrate concentration. Inhibition was a function of the identity of the salt anion within a Hofmeister (lyotropic) series: I- greater than SCN- greater than NO3- greater than Cl- greater than F-, and, in these terms, was not significantly different for a series of inorganic chlorides (Li+, Na+, K+, Cs+). The effects of salts on the natural lipoprotein substrates, chylomicrons, and very low density lipoproteins were similar to those obtained with a synthetic lipid-protein substrate complex. These findings are discussed in the light of recent ideas on the activation of lipoprotein lipase.


Phosphatidylglycerol in lung surfactant. III. Possible modifier of surfactant function.

Lamellar bodies and alveolar lavage from adult mammalian lung contain unusually high concentrations of phosphatidylglycerol that could serve as a sensitive indicator of surfactant. Phosphatidylglycerol was absent and phosphatidylinositol was correspondingly prominent in surfactant from the preterm rabbit fetus. Phosphatidylglycerol rapidly appeared and phosphatidylinositol decreased following the delivery. Surfactant isolated from the prematurely born rabbit or from humans with respiratory distress syndrome never contained phosphatidylglycerol. Comparison between lamellar bodies from fetal and postnatal rabbits revealed remarkably similar composition except for the acidic phospholipids; however, the physico-chemical properties were different. The compressibility of the surface film (i.e. the ratio of the fractional decrease in surface area and the corresponding decrease in surface tension) at low surface tensions was higher with fetal than with postnatal surfactant, whereas the difference in minimum surface tensions was small. These data suggest that phosphatidylglycerol is not an essential component required for the formation of the complex, but it improves the properties of surfactant in stabilizing the alveoli.


Gas-liquid chromatography of bile acids: a new liquid phase for both acetate and trimethylsilyl derivatives.

Polymetaphenoxylene (PPE-20) has been found to be more useful than cyclohexane dimethanol succinate (HI-EFF-8-BP) for trimethylsilyl derivatives of bile acids and to be preferable to trifluoropropyl substituted silicone (OV-210, QF-1) for analysis of their acetate derivatives.


Study of the effect of ACTH and cortisol on the plasms lipids in man using thin-layer chromatography.

This is a study of the action of ACTH and cortisol on plasma lipids using thin-layer chromatography and evaluation by densitometry. We have studied the action of 50 I.U. of ACTH in eight healthy individuals. We found an increase of free fatty acids after four hours and of triglycerides and esterified cholesterol after six hours. Free cholesterol levels declined after another six hours, the decrease becoming more intense after twenty-four hours. After twenty-four hours lecithin underwent a decline which is accompanied by an increase of lisolecithin. There was a decrease in the total amount of phospholipids. Seven patients received 100 mg. of cortisol intravenously over a period of ten minutes. We obtained an increase in free fatty acids after four hours and an increase of cholesterol after half and hour. We concluded that ACTH activates peripheral lipolysis thereby facilitating the hepatic synthesis of triglycerides and that it exerts a stimulating action on the lecithin-cholesterol-acyl-transferase system. Cortisol acts by facilitating the synthesis of triglycerides in the liver and increasing the hydrolysis of triglycerides in the plasma.


Changes in adrenocortical function in male and female guinea-pigs during maturation.

Adrenal cortisol secretion is greater in female than male guinea-pigs and declines with maturation in animals of both sexes. In an attempt to determine the intra-adrenal mechanisms responsible for age and sex influences on corticosteroid output, adrenocortical enzyme activities were compared in sexually immature (3 weeks) and mature (17 weeks) animals. Adrenal mitochondrial protein concentration decreased with maturation in male and female guinea-pigs. 11beta-Hydroxylase activity in adrenal mitochondria was also lower in mature than immature guinea-pigs but greater in males than females. Neither mitochondrial cytochrome P-450 concentration nor cholesterol side-chain cleavage activity varied with age or sex. Adrenal microsomal protein concentration and 21-hydroxylase activity were similar in male and female guinea-pigs of the same age but far greater in mature than immature animals. Microsomal cytochrome P-450 concentration was unaffected by age or sex. Adrenal delta4-steroid (cortisol) hydrogenase activity increased with maturation in both male and female guinea-pigs and was higher in males than females. These observations indicate that cortisol secretion, as modified by age and sex, correlates closely with adrenal steroid reductive but not oxidative metabolism, suggesting that changes in delta4-hydrogenase activity are responsible, at least in part, for the decline in adrenal secretion during maturation in guinea-pigs.


Endocrine and environmental influences upon plasma cortisol concentrations and plasma renin activity of the eel, Anguilla anguilla L.

The plasma concentrations of cortisol, sodium, potassium and calcium and plasma osmolarity were determined in freshwater silver eels, after intravascular injections of eel renin preparations, mammalian ACTH, mammalian angiotensin II and eel muscle extracts. Arterial blood specimens were taken before and after injection of test substances. Partially purified eel and rat renal renins gave prolonged pressor responses in intact and hypophysectomized eels and in the nephrectomized rat anaesthetized with sodium pentobarbitone. Angiotensin, but not ACTH, produced obvious pressor responses in intact and hypophysectomized eels and in eels without their corpuscles of Stannius. Hypophysectomized eels 4-8 days after operation had reduced plasma cortisol concentrations. No change in cortisol occurred in eels after removal of the corpuscles of Stannius. Eel renin preparations and ACTH gave increased concentrations of plasma cortisol 30 min after injection into hypophysectomized and intact eels. In general, the length of the renin-generated pressor response and the increased cortisol concentration were concomitant occurrences. Angiotensin injected into eels with corpuscles of Stannius removed and into hypophysectomized eels also increased cortisol levels. Control muscle extracts produced no significant changes. There were no acute changes in plasma electrolyte concentrations after the injections. Plasma renin activity measured indirectly by bioassay of angiotensin generated in vitro was more than twice as great in eels adapted to seawater than in eels in fresh water. Plasma renin activity gradually fell when eels were transferred from seawater to fresh water, and increased when the reverse transfer was carried out.


Active transport of sodium by the malpighian tubules of the tsetse fly Glossian morsitans.

Isolated Malpighian tubules of Glossina morsitans are able to transport sodium against its concentration gradient. Their rate of secretion is dependent on the sodium concentration of the bathing medium. Potassium must be present in the bathing solution for rapid secretion to be maintained, but it does not play an active role in fluid secretion. Lithium and ammonium ions are able to substitute partially for sodium, other monovalent cations cannot. Ouabain does not affect rapid secretion by Glossina tubules in vitro. Conclusions drawn from the results are incorporated into a model of Malpighian tubule cell function in this insect.


The influence of various factors on fluid secretion by in vitro salivary glands of ixodid Ticks.

1. Salivary glands of the female ixodid tick, Dermacentor andersoni, secrete fluid in vitro when bathed in a slightly modified version of the mammalian tissue culture medium 'TC 199'. 2. Rate of salivation in vitro increases with progression of feeding, but there is no comparable increase in dry weight of the salivary glands during the early phase of engorgement. Engorged ticks secreted at only 25% the rate of 90-250 mg ticks, indicating that salivary gland degeneration has already begun in the very early post-engorgement stage. 3. A salivary gland stimulating factor can be detected in the nervous system but not in other tissues. 4. Male salivary glands secrete at only 1/20th the rate of female glands. Thus males probably do not use their salivary glands as osmoregulatory organs. 5. From the uniform lack of response to ACh and uniform response to DA in 7 ixodid tick species, it is suggested that the control of salivation is similar throughout the ixodid family.


Age-related changes in cell surface antigens of preleukemic AKR thymocytes.

Thymocytes from preleukemic AKR mice aged 5-6 mo have an altered pattern of cell surface antigens. The expression of four MuLV-related antigens on the cell surface (GIX, GCSA, gp70, p30) is markedly increased in comparison to 2-mo-old AKR mice and approximates the heightened levels of these antigens found on thymic leukemia cells. H-2 and Thy-1 alloantigens also show characteristic modifications in relation to age and leukemia development. In contrast to the high Thy-1/low H-2 levels on 2-mo-old AKR thymocytes, thymocytes from 6-mo-old mice and thymic leukemia cells frequently show a low Thy-1/high H-2 surface phenotype. As thymocytes from mouse strains with a low incidence of leukemia do not show these changes, they appear to represent a stage in the conversion of normal cells to leukemia cells.


[Enzyme kinetic determination of glucose (author's transl)].

A reaction kinetic method is described for the determination of glucose in biological fluids. Glucose is oxidized to gluconic acid and NAD is reduced to NADH in the presence of glucose dehydrogenase. The rate of formation of NADH is followed in a LKB Reaction-Rate-Meter. Together with the analogue read-out, the rate of reaction is determined with a newly developed kinetic calculator. The specifications of the calculator are given. The new method has the advantage of a high rate of throughput and good reproducibility.


[Stabilization of reduced beta-nicotinamide-adenine-dinucleotide in an organic solvent (author's transl)].

NADH is surprisingly stable, providing it is stored in anhydrous organic solvents, and not, as hitherto, in aqueous solvents. The stability of NADH was investigated in solution in ethylene glycol-Tris over a period of 136 days at temperatures of 2-8 degrees C, 19-22 degrees C, 35 degrees C, 45 degrees C and 60 degrees C. For comparison, the stability of NADH in solution in distilled water was studied under similar conditions.


A kinetic method for serum 5'-nucleotidase using stabilised glutamate dehydrogenase.

A modification of a kinetic determination of 5'-nucleotidase (EC 3.1.3.5) activity is described. Special attention has been paid to the stabilisation of glutamate dehydrogenase (EC 1.4.1.2) by L-leucine, optimal NADH concentration and the influence of endogeneous ammonia. The optimal concentrations of the other constituents of the reagent were checked with the optimal values given in the literature. Normal values were determined. The proposed method shows a good correlation with a colorimetric reference method.


Adenosine 3',5'-monophosphate, morphogenetic movements and embryonic neural differentiation in Pleurodeles waltlii.

Evidence is presented for the following hypothesis dealing with dibutyryl-c-AMP induction of neural differentiation in Pleurodeles waltlii embryos: (1) Dibutyryl-c-AMP induces the determination for neural differentiation. (2) Additional dibutyryl-c-AMP is not needed for cytodifferentiation. (3) Cytodifferentiation of induced cells can only take place after morphogenetic movements (in vivo) or extensive cell migrations (in vitro). It is suggested that cell migrations, both in vivo and in vitro, lead to the separation of the induced cells from one another or from unidentified inhibitory cells.


Distribution of cytochrome-like respiration in streptococci.

The electron transport systems of 134 strains of streptococci were studied after aerobic growth on glucose in the presence of haematin, by examining the inhibition of electron transport as well as the cellular site of NADH oxidation. Each strain was placed into one of three possible groups: cytochrome-like NADH oxidase; flavin-like NADH oxidase; or no NADH oxidase. Most (88%) of the strains of Streptococcus faecalis and its variants liquefaciens and zymogenes and a few strains of S. lactis and its variant diacetylactis contained cytochrome-like respiratory systems. Other streptococci including S. faecium fell into one of the other groups but did not contain cytochrome-like NADH oxidases.


Fractionation by differential and zonal centrifugation of spheroplasts prepared from a glucose-repressed fission yeast Schizosaccharomyces pombe 972h-.

A method is described for the preparation of spheroplasts in high yield from Schizosaccharomyces pombe, by treating cells grown in the presence of glucose and deoxyglucose with snail digestive enzymes. Gentle disruption of such spheroplasts yielded homogenates, from which marker enzymes for nuclei (NAD pyrophosphorylase) and mitochondria (cytochrome c oxidase activity and spectroscopically-detectable cytochromes a + a3) could be quantitatively sedimented by low-speed centrifugation. In contrast to previous findings with Saccharomyces carlsbergensis, cytochrome c oxidase and another mitochondrial enzyme, succinate dehydrogenase, were completely sedimentable by zonal centrifugation in sucrose gradients in the presence of either 2 mM-MgCl2 or 0-4 mM-EDTA. Mitochondria were apparently smaller and of lower buoyant density in gradients containing EDTA. The bulk of the total units of malate dehydrogenase and NADH; cytochrome c oxidoreductase sedimented with mitochondria, whereas NADPH: cytochrome c oxidoreductase was located in fractions containing no mitochondria. The distributions of mitochondrial enzymes were heterogeneous in populations of mitochondria separated on the basis of size or density. The possible origins of mitochondrial heterogeneity in extracts of S. pombe are discussed with special reference to changes in the enzyme activities of cells during the cell cycle.


Biochemical and genetic characterization of a carbamyl phosphate synthetase mutant of Escherichia coli K12.

An unusual Escherichia coli K12 mutant for carbamyl phosphate synthetase is described. The mutation was generated by bacteriophage MUI insertion and left a 5% residual activity of the enzyme using either ammonia or glutamine as donors. The mutation is recessive to the wild-type allele and maps at or near the pyrA gene, but the mutant requires only arginine and not uracil for growth. By a second block in the pyrB gene it was possible to shift the accumulated carbamyl phosphate to arginine biosynthesis. The Km values and the levels of ornithine activation and inhibition by UMP were normal in the mutant enzyme.


Stability of enzymes in starving Arthrobacter crystallopoietes.

Cell-free extracts prepared from spherical and rod-shaped cells of Arthrobacter crystallopoietes were assayed for enzymes during various periods of starvation. The level of NADH oxidase dropped to 20 and 30%, respectively, in spherical and rod-shaped cells during the first 1 to 2 days of starvation and then remained constant for 9 days. Catalase activity decreased continuously and reached a low level in 9 days. Enzymes involved in glucose metabolism and the tricarboxylic acid cycle were stable for the duration of the experiment (about 1 week). Succinic dehydrogenase, fumarase and aconitase were stable during 21 days of starvation, which is the longest time enzymes have been shown to be stable in any bacterium under conditions of total starvation.


Classification of micrococci on the basis of deoxyribonucleic acid homology.

The DNA homology relationships of 25 micrococci (15 strains of Micrococcus, eight strains of Sarcina and two strains of Staphylococcus) were studied by the deoxyribonucleic acid hybridization method using nuclease S1, an endonuclease specific for single-stranded DNA molecules. Nineteen of the strains were classified into three groups. Group I contained Micrococcus lysodeikticus IAMI056, M. luteus IAMI1010, M. flavus IAMI2005 and IAMI2006, Sarcina flava IAMI2007 and IAMI1006. S. subflava IAMI2009, S. lutea ATCC381, and ATCC382, and M. luteus IAMI1006. Group II contained M. roseus IAMI315, ATCC412, ATCC185 and IAMI295. Group III contained S. lutea IAMI099, IFO3232 and ATCC383, M. varians ATCC399 and Staphylococcus lactis ATCC15306. Micrococcus luteus IAMI097, M. varians ATCC19099 and ATCC19100, M. conglomeratus IAMI459 and IAMI470, and St. aureus IAMI011 could not be assigned to any of the three groups. The grouping corresponds to that derived from the results of differential lysis by lysozyme, 'lytic enzyme 2' from Cytophaga sp., or Streptomyces albus G enzyme; and to types of peptidoglycan in the cell walls and genetic transformation. The usefulness of classification based on sensitivity to various lytic enzymes was demonstrated. Group I probably coincides with M. luteus of Bergey's Manual of Determinative Bacteriology (1974), and groups II and III with M. roseus and M. varians respectively.


Permissiveness of mouse, monkey and hybrid cells to encephalomyocarditis (EMC) virus.

Encephalomyocarditis (EMC) virus replicates to high titre in permissive mouse kidney (MKS) cells but poorly in monkey kidney (CV1) cells. The permissiveness of monkey-mouse hybrid cells varies according to their chromosomal content. In monkey cells, the synthesis of both single-stranded and double-stranded virus RNA is restricted; in semi-permissive hybrid clones, the double-stranded RNA is synthesized normally, whereas the synthesis of the single-stranded RNA is inhibited. Thus, it seems that more than one restrictive event is responsible for the low permissiveness of monkey cells to EMC virus.


Enhancement of polysome formation and protein synthesis in Sendai virus-infected cells.

A considerable stimulation of total protein synthesis in Sendai virus-infected chicken fibroblasts occurred 18 h after infection. The stimulation of synthesis was shown to be due mainly to the synthesis of virus-specific proteins; the synthesis of host proteins was either unchanged or slightly enhanced. The level of functional polyribosomes in virus-infected cells was also increased as compared to that in mock-infected control while the number of free 80S ribosomes was proportionally decreased.


Ribonucleotides in infectious bovine rhinotracheitis virus DNA.

Infectious bovine rhinotracheitis (IBR) virus was grown in the presence of 5-3H-uridine in a continuous line of bovine kidney cells. 5-3H-uridine was found to be associated with viral nucleocapsids. Furthermore, purification of the viral nucleic acid present in nucleocapsids illustrated that 5-3H-uridine was part of the viral nucleic acid. Purification of viral DNA from infected cells also indicated that 5-3H-uridine was associated with viral nucleic acid possibly as ribonucleotides. The label was identified as RNA by measuring its susceptibility to RNase and analysis of the bases. Short pulses with 5-3H-uridine, resulted in labelled nucleic acid which was extremely sensitive to RNase and alkali but resistant to DNase. Nucleotide analysis indicated that after short pulses all the radioactivity was associated with the base uracil whereas upon longer labelling periods a large percentage of the label was associated with cytosine. However even if viral DNA was isolated from nucleocapsids there was still some radioactivity associated with uracil. Sedimentation of heat denatured 5-3H-uridine label viral nucleic acid in CS2SO4 indicated that the label sedimented at a density of single stranded DNA suggesting that the ribonucleotides are covalently linked to the viral DNA.


Rescue of a rat tropic rat hepatoma virus pseudotype Kirsten sarcoma virus by co-cultivation of hepatoma tissue culture cells with K-NRK cells.

A rat tropic rat hepatoma virus pseudotype Kirsten sarcoma virus has been rescued from a co-culture of rat hepatoma tissue culture cells, HTC-HI, with Kirsten murine sarcoma transformed non-producer rat kidney cells (K-NRK). The virus complex, RHHV-KiMSV, released from the co-culture exhibited cell transformation ability on normal rat kidney (NRK) cells and showed a restricted host range limited to rat embryo fibroblasts and other normal rat cell lines. Evidence derived from indirect immunofluorescence assays demonstrated the association of rat helper virus specific gs-I and gs-3 antigens with RHHV-KiMSV transformed cells. RHHVKiMSV is currently being cultured at a titre as high as 10(4) f.f.u./ml in tissue culture, and thus offering opportunity for biochemical studies of rat viruses.


Characterization of two particle types of calf rotavirus.

Two distinct types of rotavirus particle were isolated in caesium chloride density gradients. The higher density particle banded at 1-38 g/ml and measured 55 +/- 0-4 nm in diameter while the less dense particle banded at 1-36 g/ml, measured 66 +/- 0-4 nm in diameter and appeared to possess an extra outer capsid layer. Some forms intermediate between the two types were seen. Since the two particles had a similar but not identical morphology and polypeptide composition, they were considered to be different forms of the same virus. Infectivity was associated with the larger particle which contained one more polypeptide than the smaller particle.


Human fibroblast and leukocyte interferons show different dose-response curves in assay of cell protection.

Preparations of human fibroblast and leukocyte interferons of similar potency show markedly different dose-response curves in an assay which measures the degree of protection of tissue cultures against virus c.p.e. The effect is observed with both vesicular stomatitis virus (VSV) and Mengovirus, and is not altered by purification of the interferons.


Requirement of an adenylic acid-rich segment for the infectivity of encephalomyocarditis virus RNA.

About 80% of the RNA molecules extracted from encephalomyocarditis (EMC) virus were bound by oligo(dT)-cellulose under conditions which bind poly(A) but not poly(C) nor ribosomal RNA. This shows that most EMC virus RNA molecules contain a poly(A) tract. Both bound and unbound fractions contained RNA molecules of apparently the same size when examined by sucrose gradient sedimentation, but the bound fraction contained an adenylic acid-rich segment of about 20 nucleotides long, whereas the unbound RNA did not. The bound RNA had 200 times the specific infectivity of the unbound RNA which suggests that the poly(A) tract present in EMC virus RNA is required for infectivity.


Herpes simplex virus specified deoxypyrimidine kinase and the uptake of exogenous nucleosides by infected cells.

Herpes simplex virus can confer to thymidine kinaseless cells the ability to incorporate exogenously supplied thymidine into acid precipitable material. However no incorporation of exogenously supplied deoxycytidine into acid precipitable material can be detected after infection of deoxycytidine kinaseless cells by herpes simplex virus. This failure to incorporate exogenous deoxycytidine is not due to the failure of the deoxycytidine phosphorylating activity of the virus induced deoxycytidine kinase but to a block in the metabolism of deoxycytidine monophosphate in herpes simplex virus infected cells. This block becomes evident with the appearance of the virus induced deoxypyrimidine kinase activity.


The cleavage of polyoma virus DNA by restriction enzymes KpnI and PstI.

The action of two restriction endonucleases on polyoma virus DNA has been examined and the sites at which they cleave the DNA located. One of the enzymes, KpnI from Klebsiella pneumoniae OK8, cleaves polyoma DNA twice at about 11-6 and 59-2% from the EcoRI site. The other enzyme, PstI from Providencia stuartii 164, cleaves polyoma DNA five times at about 14-8, 16-5, 32-6, 50-3 and 80-0% from the EcoRI site. Some of the cleavages produced by these enzymes alone, or in conjunction with other endonucleases, may be of use in the isolation of regions of particular interest from the virus DNA.


Use of a transfection method to demonstrate a monolayer cell transforming agent from the EB3 line of Burkitt's lymphoma cells.

Primary human amnion cell monolayers which had been treated with DEAE-dextran, washed, and then inoculated with sonicated cells of the EB3 line of Burkitt's lymphoma cells developed foci of transformed amnion cells 7 to 14 days later. When either the DEAE-dextran or the sonicate was omitted, no significant transformation was found. The foci consisted of enlarging mounds of rapidly dividing cells, which upon subculturing continued their high miotic activity; and strains or lines of the transformed amnion cells were thus readily established. The modal number of chromosomes in such lines was 65 instead of the normal 46. Not all human amnions yielded cells transformable by EB3 cell sonicate, as determined by direct comparisions using the same cultural conditions and testing with the same fresh  sonicate preparation in the same experiment. Overall, it appeared that only about 40 to 50% of the amnions yielded transformable cell monolayers; the rest gave monolayers apprently completely  refractory to the transformation. The transformed amnion cells contained nuclear and cytoplasmic Epstein-Barr virus (EBV) antigen(s), as revealed by indirect immunofluorescence tests. EB3 cell sonicate also caused the appearance of rapidly growing transformed cell foci on secondary rat embryo cell  monolayers which had been sensitized with DEAE-dextran. Calcium in  the cell maintenance medium decreased the number of transformed foci found, both on the human and on the rat cell monlayers. Sonicates of cultured  normal human leucocytes had no such transforming activity for either the human or the rat cells. The transforming agent in EB3 cell sonicate was completely destructible by either deoxyribonuclease or trypsin, but not by ribonuclease, and was not neutralizable by anti-EBV serum. The simplest interpretation of these results is that the transforming agent is part of all of the EBV DNA plus some necessary protein, with both the DNA and the protein accessible to hydrolytic enzyme action.


Control of protein synthesis in herpesvirus-infected cells: analysis of the polypeptides induced by wild type and sixteen temperature-sensitive mutants of HSV strain 17.

The polypeptides induced in cells infected with a Glasgow isolate of HSV-I (17 syn+) have been characterized by SDS polyacrylamide gel electrophoresis. Study of the kinetics of synthesis in three cell lines has detected a total of 52 polypeptides, 33 of which can be identified in polypeptide profiles of purified virions. These include six low mol. wt. polypeptides that have not been previously reported. Several polypeptides were labelled with glucosamine in infected BHK cells. The different polypeptide patterns obtained at permissive (31 degrees C) and nonpermissive (38 degrees C) temperature in cells infected with 16 temperature-sensitive (ts) mutants are reported. The effect of multiplicity of infection (m.o.i.) on the polypeptide profile has been examined for two of the DNA -ve mutants: below ten, the profile varied with the m.o.i. whereas above ten it was constant. All mutants were therefore examined at an m.o.i. of approx. 20. Mutants from the same complementation group showed very similar profiles. A number of general conclusions concerning control of protein synthesis in HSV infected cells can be made: (I) As most of the 16 ts mutants affected the synthesis of several or many polypeptides it follows that a large proportion of genome specifies controlling functions. (2) The high frequency with which some polypeptides were affected suggests they are at or near the terminus of biosynthetic pathways which are under multiple control. (3) Conversely, some polypeptides were affected with a low frequency suggesting that their synthesis is not dependent on the expression of many virus functions. (4) Several individual ts mutations lead to the synthesis of increased amounts of different large polypeptides. (5) Analysis of every band detectably affected by at least one ts mutation has disclosed nine classes of dependence relationship between polypeptide synthesis and the DNA phenotype of the mutants, illustrating that this relationship is complex and different for different polypeptides. (6) The inhibition of host protein synthesis by the virus may not be a simple single step process.


Inhibition of murine sarcoma virus induced transformation by adenovirus structural proteins.

The purified fibre and hexon of adenovirus 12 inhibit the transformation in tissue culture of murine sarcoma virus (msv-m) by as much as 80% and 70%, respectively, when they are added to cells 8 to 2o h before MSV-M infection. During a 12 h period, only about 6 to 8% of added radiolabelled viral proteins become associated with cells (or I-O mug protein bound/10(5) cells). No inhibition occurs when the proteins are added simultaneously with MSV-M or 90 min or 4 h after MSV-M. There is also a direct correlation between the extent of focus inhibition and the concentrations of viral proteins used. Concentrations of viral proteins used to inhibit cell transformation do not affect cell growth, but do reduce cell macromolecular synthesis.


Demonstration of a core in poliovirus particles by electron microscopy.

Electron microscopic examination of poliovirus in CsCl-solutions without conventional staining with phosphotungstic acid or uranyl acetate revealed the existence of cores inside poliovirus particles. Empty capsids could not be visualized by this technique. Evidently, CsCl can enhance the contrast of the virus particle to an extent sufficient for the study of poliovirus particles by electron microscopy under conditions in which they exhibit specific properties.


Some electrical properties of motor units and their effects on the methods of estimating motor unit numbers.

Long train (50) electrical pulses were applied to the ulnar, median, and deep peroneal nerves. The probability of firing, or firing index, of the first motor units (MU) recorded by surface and needle electrodes in the first dorsal interosseous, thenar, and extensor digitorum brevis muscles varied from 0-100% as the stimulus was increased from a minimum threshold voltage (V) to V + delta V (delta V = 2-4 volts). The voltage interval, delta V (firing level range), of even the first few MU recruited greatly overlapped in normal subjects, but may overlap less in some neuromuscular disorders such as amyotrophic lateral sclerosis. The mean size of the single MU isolated by (1) isometric voluntary contraction method, (2) F recurrent discharge method, and (3) stimulation at multiple locations along the nerve, were more than twice as large as the mean size of the incremental steps evoked by graded electrical stimulation. These results suggest that methods of estimating the number of MU in a muscle should incorporate a correction for the fluctuations in excitability and overlap in firing levels of MU, and in addition should include larger MU in the estimation of the mean MU potential.


Sensory and motor residual latency measurements in helathy patients and patients with neuropathy-part 1.

Median and ulnar nerves were evaluated using established nerve conduction techniques in a control population and in a population of patients with neuropathy. Motor and sensory residual latencies were obtained. The main contributor to the latencies was slowing of the nerve conduction in the smaller attenuated nerve fibres of the hand and fingers.


Neurological disorders in the elderly at home.

Eight hundred and eight subjects participated in three surveys of random samples of people aged 65 years or more living in their own homes. Neurological history and examination showed the prevalence of completed stroke to be 73 per 1000. Eighty-seven subjects per 1000 gave a history of transient cerebral ischaemic attacks. These prevalence rates were unaffected by age or sex. Senile dementia was diagnosed in 24 subjects per 1000 under 75 years and 109 per 1000 over that age. The prevalence of dementia of all types was 43 per 1000 under, and 140 per 1000, over 75 years of age. Parkinsonism was diagnosed in 16 subjects per 1000, and essential tremor in 17 per 1000. The prevalence of epilepsy was four subject per 1000. Other neurological disorders were diagnosed in 36 subjects, and a similar number had neurological abnormalities to which a definite diagnosis could not be given.


Metronidazole neuropathy.

Two patients are described who developed sensory neuropathy after the ingestion of 30.6 and 114 g metronidazole respectively. The drug, widely used in gastroenterological and gynaecological practice, has not hitherto been considered neurotoxic. The implications are stressed in relation to the differential diagnosis of patients with gastroenterological disorders who develop peripheral neuropathy.


Diencephalic amnesia: a reappraisal.

The anatomical basis of memory disorder related to lesions of the diencephalon is a controversial matter. A study of a patient who developed severe amnesia in association with bilateral metastatic tumour invasion of the medial and posterior thalamus is reported. The findings in this case have led to further discussion of this unsettled issue.


Muscle wasting after prolonged hypoglycaemic coma: case report with electrophysiological data.

A case of distal muscle wasting associated with hypoglycaemia is described. Motor and sensory conduction studies are reported that provide little evidence of a hypoglycaemic peripheral neuropathy. Damage to anterior horn cells or motor roots appears to be a more likely explanation for the amyotrophy. Recovery occurred clinically with evidence of reinnervation by axonal collateral sprouting.


Bulbocavernosus reflex in normal men and in patients with neurogenic bladder and/or impotence.

The bulbocavernosus reflex was investigated electrophysiologically in 14 normal adult male subjects and in 80 patients with neurogenic bladders and/or impotence due to various neurological causes as well as in patients with functional impotence. The glans penis was stimulated superficially by single electrical shocks and the reflex responses were recorded from the bulbocavernosus (BC) and the striated anal sphincter muscles by means of concentric needle EMG electrodes. In all normal subjects, the BC reflex was recorded from the BC muscle as a stable and constant response having a mean latency of 36.1 msec. A response from the external anal sphincter was obtained in only 21% of the subjects investigated. In 13 patients with cauda equina lesions, the BC reflex was either absent or was present with a prolonged latency. Twenty-two patients with polyneuropathy of various causes were also investigated; in these patients the latency of the BC reflex was significantly greater than in the normal controls, but the most abnormal results were obtained in cases of alcoholic polyneuropathy. In the 19 cases of spinal cord disease with spasticity the BC reflex response was very intense, often with after discharges but latency values were within normal limits. In the 16 cases with functional impotence, the BC reflexes were basically normal; but in 3 cases, the threshold of the reflex was significantly raised, and in 1 case a prolonged latency was observed. The value and the practical application of the BC reflex in the BC reflex in the differential diagnosis of bladder dysfunction and of impotence was stressed.


Antibodies to Epstein-Barr virus in neurological diseases.

An association of evidence of Epstein-Barr virus (EBV) infection in 3 different cases of neurological disease not related to infectious monomucleosis is reported. A rise in antibody titres to EBV-viral capsid antigen (VCA) and early antigen (EA) in the serum and CSF was demonstrated in a patient with acute encephalitis and in a patient with a psychotic-like organic brain syndrome. Both patients demonstrated albuminocytological dissociation in the cerebrospinal fluid. IgM antibodies specific to EBV-VCA indicating a primary infection by EBV were found in both patients. In the second, concomitant infection with HSV1 probably preceding EBV infection was found. In the third patient with polyneuritis, elevated antibody titres to EBV-VCA were demonstrated. No evidence of penetration of antibodies to EBV through the blood-brain barrier, resulting from the elevated level in the serum, was demonstrated.


In vivo reversal of thyroxine induction of DNA synthesis by dibutyrly cyclic AMP in developing rat cerebellum.

Thyroxine, dibutyryl cyclic AMP, and a combination of both drugs were administered daily from birth to 2, 2 and 3 pups, respectively from each of 5 litters of Sprague-Dawley rats. Body weight, brain weight, cerebellar weight, and cerebellar DNA were measured in each animal at age 5 days and compared with values from a pair of controls from each litter. Cerebellar weight and DNA content were affected more severely than body weight in cyclic AMP-treated animals, with cerebellar DNA reduced significantly to 88% of control values. Cerebellar DNA was significantly elevated to 117% of control values in thyroxine-treated animals. This augmentation of cerebellar DNA synthesis by thyroxine was negated by administration of dibutyryl cyclic AMP 10 min prior to the thyroxine injection. These results support an hypothesis that the enhancement of cerebellar cell division by thyroxine involves an increase in the ratio of intracellular cyclic guanosine monophosphate to cyclic adenosine monophosphate. The reversal of the thyroxine-induced increase in cerebellar DNA synthesis by a prior injection of dibutyryl cyclic AMP suggests that the early stimulation of cell division by thyroxine may be mediated by cyclic AMP, and that the intracellular balance between cerebellar cyclic AMP and cyclic GMP was distorted by in vivo elevation of intracellular cyclic AMP levels.


Hereditary motor peripheral neuropathy predominantly affecting the arms.

A kinship is described in which there was slowly progressive wasting and weakness of the muscles of the upper and occasionally of the lower limbs. Some members had hyperreflexia. There were no sensory abnormalities. Electrophysiological study suggested the presence of motor peripheral polyneuropathy. The condition appeared to be inherited as an autosomal dominant. The disorder does not appear typical of any of the known hereditary polyneuropathies and it is possible that it may represent a unique hereditary, dominantly motor, polyneuropathy. The significance of the hyperreflexia is uncertain, but raises the possibility of minor central involvement as well as peripheral neuropathy.


Pigmented ganglioneuroblastoma: relation of melanin and lipofuscin to schwannomas and other tumors of neural crest origin.

An unusual case of ganglioneuroblastoma containing melanin is presented. Electron microscopy revealed various stages of development of melanosomes in neoplastic cells of Schwann, the first direct demonstration in human material that these cells are malanogenic. The frequent occurrence of neuromelanin in autonomic ganglia and in ganglioneuromas is interpreted as the presence of altered lipofuscin. Review of ultrastructural and other observations indicates a relation between various pigmented tumors, the cell of Schwann, and other cells arising from the neural crest.


Gian axonal neuropathy--a generalized disorder of cytoplasmic microfilament formation.

Light and electron microscopy of a sural nerve biopsy obtained from a patient with giant axonal neuropathy revealed the presence of numerous axonal spheroids which were composed chiefly of neurofilaments. Large discrete masses of cytoplasmic microfilaments were also observed in endoneurial fibroblasts, endothelial cells, Schwann cells, perineurial cells and cultured skin fibroblasts. Neuronal degeneration in giant axonal neuropathy appears to be part of a generalized disorder of cytoplasmic microfilament formation.


Melanoma of the pituitary. Case report.

A case is described in which a pigmented tumor occupied the pituitary fossa with a large suprasellar extension. Its histological and ultrastructural appearances resembled those of a melanoma. The origin of the tumor is discussed with particular reference to Rathke's pouch.


Pathogenesis of aqueductal occlusion in congenital murine hydrocephalus.

In a study of congenital hydrocephalus in the murine mutant (hy-3/hy-3), the authors found that aqueductal stenosis develops during the progression of hydrocephalus. In Stage 1 hydrocephalus (ventricular dilation and open aqueduct), a block in the subarachnoid space over the cerebral convexities causes the lateral and third ventricles to enlarge. The ependyma becomes stretched and a collection of edematous fluid forms in the subependymal layer. In Stage 2 hydrocephalus (edema in white matter around lateral ventricles and compression of quadrigeminal plate), edema develops peripheral to ependyma in the aqueduct and compresses the lateral surfaces of the aqueductal wall to obstruct the lumen. While periaqueductal edema is spreading, the forces of the expanding midline structures and the cystic occiital horns alter the relationship of brain structures. There is no proliferation of glia, but, rather, a "simple stenosis" which results from a combination of ventricular dilation, cerebral edema, brain shift, brain-stem compression, and brain-stem edema. In this study, normal ependymal specializations were observed that indicate a more active functional role for aqueductal ependyma than previously recognized.


Relationship between bone uptake of 99mTc-pyrophosphate and hydroxyproline in blood and urine.

In a group of hospital patients with various diseases, the urinary hydroxyproline-to-creatinine ratio showed a significant correlation (r = 0.63; p less than 0.001) with the 5-hr bone-to-soft-tissue ratio for 99mTc-pyrophosphate uptake. In patients on chronic hemodialysis, a similar correlation was found between the 5-hr bone-to-soft-tissue ratio and hydroxyproline levels in plasma and serum. The findings suggest that 99mTc-pyrophosphate binding by bone is related to collagen metabolism.


Carboxyl-labeled 11C-1-aminocyclopentanecarboxylic acid, a potential agent for cancer detection.

Carboxyl-labeled 11C-1-aminocyclopentanecarboxylic acid (11C-ACPC) has been prepared in multimillicurie amounts. The conversion of H11CN to 11C-ACPC (t1/2 = 20.4 min) was accomplished by a rapid (20-min) two-step high-temperature modification of the Bücherer-Strecker amino acid synthesis technique, which should be applicable at other accelerator installations. Purification was by ion exchange techniques. Animal studies have indicated that 11C-ACPC is a potential tumor-localizing agent for detecting cancer in humans by nuclear medicine scanning techniques.


Ventilation and sleep states in newborn infants.

Recent studies have shown that ventilation in the newborn period is affected by sleep state. We investigated various measures affecting ventilation using the single breath airway occlusion technique in ten healthy, full-term newborn infants. There was a significant increase in respiratory rate and in minute ventilation in rapid eye movement sleep compared to non-REM sleep, and there was no significant change in tidal volume between the two sleep states. The variability of ventilation was increased in REM sleep, and inspiratory pressure at one-tenth of a second following airway occlusion was significantly increased in REM sleep. Effective elastance was similar in both sleep states. Measures that reflect activity of the Hering-Breuer reflex were significantly increased in non-REM sleep as compared with REM sleep. These results document the interdependence of sleep state and respiratory control mechanisms in full-term infants.


Respiratory syncytial virus infections in infants: quantitation and duration of shedding.

Infants hospitalized with respiratory syncytial virus infection were studied to delineate the quantitative shedding patterns and duration of shedding of RSV. Nasal wash specimens collected daily from 19 infants contained a mean maximal titer of 4.34 log10 50% tissue culture infective doses per milliliter. On admission, the mean titer was 4.14 log10 TCID50, with no consistent decline until after Day 6. The mean duration of shedding for 23 patients until they were virus negative was 6.7 days with a range of 1 to 21 days. Quantities of RSV shed were significantly greater in infants less than one month of age and in infants with evidence of pulmonary consolidation on chest roentgenogram. Shedding extended for a significantly longer time in infants with lower respiratory tract disease than in those with clinical manifestations limited to the upper respiratory tract.


Protracted pneumonitis in young infants associated with perinatally acquired cytomegaloviral infection.

Two infants developed a protracted pneumonitis with lower respiratory obstruction beginning at one month of age. Lung biopsy in one suggested a viral etiology which prompted an extensive investigation of each infant for specific etiology. Virologic, serologic, immunologic, and electronmicroscopic studies indicated that cytomegalovirus was a major causative factor. The infections were apparently acquired at birth from infected maternal genital tracts and have persisted for prolonged periods of time. Evidence for gross immunologic defect as a precipitating cause was lacking. These infants serve to emphasize the possible pathologic potential of CMV when acquired in early life even in the absence of iatrogenic immunosuppression.


Long-term hepatocellular effects of hycanthone and of two other anti-Schistosomal drugs in mice infected with Schistosoma mansoni.

In confirmation of an earlier study, it was found that in mice infected with Schistosoma mansoni hepatocellular carcinomas are induced by hycanthone, an antischistosomal drug and potent frameshift mutagen. In addition, a high incidence 1) of micronodular hepatocellular whirling lesions with increased basophilia, 2) of other proliferative areas of altered cellularity and 3) of precancerous nodules was found in the livers of schistosome-infected mice treated with hycanthone. A dosage schedule of hycanthone which was too small to have any significant chemotherapeutic effect in mice (3 X 3 mg/kg) was sufficient to induce a statistically highly significant incidence of micronodular lesions and of precancerous nodules. Hence, the hepatic tissue proved more susceptible to low doses of this drug than the parasite. No significant hepatocarcinogenic effects or other hepatocellular changes were induced by chemotherapeutically effective doses of two related equipotent antischistosomal compounds, a chloroindazole analog of hycanthone, IA-4, and the tetrahydroquinoline derivative, oxamniquine.


A study of the effects of p-chloromercuribenzene sulfonic acid on acetylcholine-induced responses of molluscan neurons.

The effects of the organic mercurial p-chloromercuribenzene sulfonic acid (PCMBS) on the acetylcholine receptors of two types of neurons from the marine gastropod Anisodoris nobilis were studied. Cell types were distinguished by the ion-specific conductance change resulting from applied cholinergic agents: in certain cells a conductance increase to Cl resulted (Cl-neurons); in other cells a cation conductance increase resulted (D cells). Exposure of the Cl-neurons to 1 mM PCMBS at 4 degrees C had two distinct effects: 1) the sensitivity of the membrane conductance to microelectrophoretically applied carbamylcholine was blocked in a few minutes; and 2) the membrane conductance slowly increased with an inversion potential which was the same as that of the cholinergically induced response. d-Turbocurarine did not appreciably reduce this conductance increase. However, applying carbamylcholine during the exposure period to PCMBS reduced the PCMBS response. The cholinergic response of D cells was affected to a much lesser degree by PCMBS.


Electrophysiological observations on the action of the purified scorpion venom, tityustoxin, on nerve and skeletal muscle of the rat.

When tityustoxin (TsTX, 1.43 muM; mol. wt. 6995) was applied to the rat phrenic nerve-diaphragm muscle preparation, both the directly and indirectly elicited muscle contractions were transiently potentiated by about 30 and 60%, respectively, and the half-relaxation time was prolonged to more than 10 times the control. The onset of potentiation of muscle contraction was immediate and coincided closely with the prolongation of the directly elicited action potential and depolarization of the muscle membrane but preceded the presynaptic effect of the toxin. TsTX antagonized the neuromuscular blockade produced by low concentrations of d-tubocurarine (6.5 muM) but not by higher concentrations of d-tubocurarine (13 muM) or by alpha-bungarotoxin (5 mug/ml). Single shocks to the nerve of toxin-treated muscles evoked repetitive end-plate potentials which summated and triggered action potentials and muscle contractions 15 minutes after exposure. After 25 minutes, the frequency of spontaneous transmitter release transiently increased from 2 to 550 sec-1 but was never blocked and there was no blockade of the acetylcholine receptors of chronically denervated muscle. Both the increase in miniature end-plate potential frequency and postsynaptic depolarization induced by TsTX could be blocked by tetrodotoxin, and the effects on end-plate potentials and action potentials could be blocked by lowering the external sodium concentration. Removal of calcium and addition of ethylene glycol bis (beta-aminoethyl ether)-N,N'-tetraacetic acid (EGTA, 10 mM) to the bathing fluid blocked the presynaptic but not postsynaptic effects of TsTX at 23 degrees C; at 37 degrees C the presynaptic effects of TsTX in a calcium-free medium with EGTA were depressed but still present. Delayed rectification and membrane electrical characteristics were unaffected by TsTX. The results indicate that TsTX depolarizes the pre- and postsynaptic membranes by increasing sodium permeability. The postsynaptic site of action is most likely to be the sarcolemmal membrane where TsTX delays Na inactivation and thus prolongs the active phase of muscle contraction. Presynaptically, TsTX has two sites of action: 1) it depolarizes the nerve terminal thus facilitating the spontaneous release of transmitter; and 2) it acts at the membrane of the unmyelinated nerve terminal arborizations where it prolongs the sodium current thus giving rise to a repetitive response to single shocks applied to the nerve.


Effects of ethacrynic acid and furosemide on isolated rat kidney mitochondria: inhibition of electron transport in the region of phosphorylation site II.

Previous reports that ethacrynic acid and furosemide inhibit the respiration of isolated mitochondria suggested a direct action of these diuretics on oxidative metabolism. To explore this possibility further, the effects of ethacrynic acid and furosemide on the oxygen consumption of mitochondria isolated from the cortex and outer medulla of rat kidneys were investigated. Both diuretics inhibited state 3 and uncoupled respiration supported by glutamate-malate (which enters the electron transport chain prior to site I) and succinate (which enters prior to site II); respiration supported by tetramethyl phenylene diamine-ascorbate (which enters prior to site III) was relatively unaffected. Biochemical bypass of site II significantly alleviated the respiratory inhibition by both agents. Confirmation of these findings was provided by measurement of the electron transport carriers by dual wavelength spectroscopy, which showed that both diuretics caused a reduction of flavoproteins and an oxidation of the cytochromes. It is concluded that ethacrynic acid and furosemide inhibit oxidative phosphorylation in vitro by inhibiting electron transport through phosphorylation site II.


The effect of parathyroidectomy and large doses of cholecalciferol on the ability of rats to adapt to changes in dietary intake of calcium.

1. Adaptation to different dietary levels of calcium was produced by feeding a low (0-2%) calcium diet or one of two high (1-6 or 0-8%) calcium diets for 4 or 6 weeks. Adaptive changes in true and apparent absorption of calcium, apparent absorption of phosphate and urinary excretion of calcium, were observed. 2. Parathyroidectomy performed prior to adaptation did not greatly impair the ability of rats to adapt to different levels of calcium in the diet. The response of the rats to parathyroidectomy was affected by their subsequent dietary history. 3. Six weeks after parathyroidectomy the plasma calcium was significantly higher than it had been immediately post-operatively. This rise in plasma calcium was seen in the rats adapted to the 1-6% calcium diet but not in those adapted to the 0-2% calcium diet. 4. Parathyroidectomy performed after adaptation had taken place did not abolish the adaptive changes. The response of rats to parathyroidectomy was affected by their previous dietary history. 5. Large doses of cholecalciferol given for 8 days after adaptation had taken place increased the absorption of calcium in rats adapted to the 0-8% calcium diet thereby abolishing or reducing the adaptive differences in absorption between these rats and rats adapted to the 0-2% calcium diet. The cholecalciferol increased urinary calcium excretion but did not abolish adaptive differences in urinary excretion of calcium between rats adapted to diets with different calcium levels. 6. It is concluded that parathyroid hormone does not play a major role in mediating adaptation to different dietary intakes of calcium. The possible role of 1-25 dihydroxycholecalciferol is discussed.


Synaptic transmission in parasympathetic ganglia in the urinary bladder of the cat.

1. Electrophysiological techniques were used to study the sacral para-sympathetic input to pelvic ganglia located on the surface of the urinary bladder of the cat. 2. Synaptic transmission in pelvic ganglia was mediated primarily via nicotinic receptors although muscarinic excitatory receptors were present. 3. The most prominent characteristic of transmission in pelvic ganglia was the marked recruitment elicited by increasing frequencies of preganglionic nerve stimulation. Post-ganglionic action potentials were of low amplitude at low frequencies of stimulation (0-1-0-5c/s), but commonly increased to five to twenty times control amplitudes during continuous stimulation at frequencies between 5 and 10c/s. Thus, it is proposed that vesical ganglia may act as "filters" in the micturition pathway; blocking the excitatory input to the bladder when intravesical pressure and parasympathetic firing is low and facilitating the neural input to the bladder during micturition when preganglionic activity is high. 4. Information was also obtained about the characteristics of the parasympathetic post-ganglionic neurones innervating the bladder. Stimulation of the preganglionic fibres in the pelvic nerve elicited a bimodal contraction consisting of an initial phasic response, which was atropine-resistant and a tonic response which was blocked by atropine. This suggests that two types of neurones, cholinergic and non-cholinergic, may mediate the sacral input to the vesical smooth muscle.


[Fetal and neo-natal development of brown adipose tissue in guinea pigs and rats. Feto-maternal or milk transfer of essential fatty acids : lipogenesis and morphology (author's transl)].

Brown adipose tissue (BAT) lipogenesis (fatty acid, glycerol and CO2 synthesis) and its morphology determined by optical microscopy, were studied in guinea pigs and rats during intra-uterine life and during the suckling period. Following the receptor induction and after the commencement of the hormone sensitive adenylate-cyclase/lipase system (i.e. on the 60th day in guinea pigs, on the 20th day in rats), the fetal BAT releases fatty acids (NEFA) and is capable of allowing the non-shivering thermogenesis. When the maternal diet and, consequently, the fetal or neonatal BAT are supplied with considerable linoleic acid, NEFA contain a large proportion of essential fatty acids. In vitro, the greater the linoleic acid concentration in these NEFA, the less inhibited is the lipogenesis from (2-14C) pyruvate. Thus, in periods just preceding or succeeding birth, fatty acid and glycerol synthesis are higher when the feto-maternal and/or the milk supply are enriched in linoleic acid than when they contain a large proportion of endogenous fatty acids. Morphological studies indicate that the adipose cell evolution could be nonidentical in BAT more or less enriched in essential fatty acids. Linoleic enriched BAT (of animals born to females kept on a sunflower oil diet) seemed to be in a healthy physiological state at birth, perhaps due to rapid lipid renewal and synthesis in their membranes. The control BAT (of animals born to females kept on a lard diet) appeared loaded with fats and in a worse conservation state at the same age.


Inhibition by theophylline of phagocytosis in Acanthamoeba castellanii.

Theophylline inhibited the phagocytosis of latex beads by Acanthamoeba castellanii (Neff). The cells recovered the ability to engulf beads after 1-2 hr of exposure to theophylline. Cells which have been exposed to 25 mM theophylline for a period of inhibition and recovery were not inhibited further by incubation with a fresh medium containing the same concentration of theophylline. However, the medium in which the cells recovered was as effective as a fresh medium in inhibiting phagocytosis in a fresh batch of cells, suggesting that the development of insensitivity to theophylline inhibition resides with the cells themselves. Dibutyryl cyclic AMP also inhibited bead uptake.


Genetic markers in atherosclerosis: a review.

There is a growing number of lipoprotein markers recognized by immunological, electrophoretic, and other biochemical methods, and a beginning has been made on studying their modes of inheritance and linkage relations. Suggestive but inconclusive evidence of a relation between the cerumen polymorphism and arteriosclerosis has been published. Associations of the ABO blood groups with cardiovascular disease and serum lipid levels have been established, but the exact relation to lipoproteins and atherosclerosis remains to be determined.


Trisomy 8 restricted to cultured fibroblasts.

In the course of re-examing cultured fibroblasts stored in liquid nitrogen from a patient with developmental retardation, solitary left kidney, and Wilms tumour, a cell line trisomic for chromosome 8 was found. Trisomy 8 was restricted to fibroblasts in the first 22 subcultures and was absent in later passages as well as in lymphocytes. A familial pericentric inversion of chromosome 2 was observed in three generations including the propositus but was though to be unrelated to the clinical problem. Multiple spontaneous chromosomal rearrangements were seen in several late subcultures.


The isolation of reovirus-like agents (rota-viruses) from acute gastroenteritis of piglets.

Isolations of reovirus-like agents (rotaviruses) were made from nine of 23 outbreaks of piglet diarrhoea on different farms and from both weaned and unweaned piglets. The viruses were shown to be morphologically and anti-genically similar to the rotaviruses of children and calves. Gnotobiotig piglets given intranasal inoculations of five different isolates developed acute gastroenteritis, and the virus was re-isolated from the faeces or intestinal contents. The piglet virus was not adapted to replicate in cell culture. We conclude that the pig rotavirus is commonly associated with outbreaks of gastroenteritis and is probably an important aetiological factor in this disease.


Fusion of phospholipid vesicles reconstituted with cytochrome c oxidase and mitochondrial hydrophobic protein.

Reconstituted cytochrome oxidase liposomes were fused with liposomes reconstituted with mitochondrial hydrophobic protein, which acts as a membrane-bound uncoupler of cytochrome oxidase. Fusion was assayed by the loss of respiratory control of cytochrome oxidase as measured by the increased rate of ascorbate oxidation induced by hydrophobic protein when both proteins shared the same vesicles. Fusion was dependent on the presence of phosphatidylserine in the liposomes Ca++ in the aqueous medium. Phosphatidylcholine-phosphatidylserine liposomes required higher concentrations of phosphatidylserine and Ca++ than did phosphatidylethanolamine-phosphatidylserine liposomes. Cytochrome oxidase vesicles containing high concentrations of phosphatidylserine showed little or no respiratory control, while those with lower concentrations showed high respiratory control; respiratory control could be induced by fusing cytochrome oxidase vesicles containing high phosphatidylserine with protein-free liposomes containing low phosphatidylserine concentration. If cytochrome oxidase vesicles and hydrophobic protein vesicles were prefused separately for 15 min, they lost the ability to fuse upon being subsequently mixed together. The reconstituted vesicles had diameters of about 200 A; fusion yielded vesicles with diameters in excess of 1000 A.


Oildroplets in the eyes of adult anuran amphibians: a comparative survey.

Oildroplets in the eyes of terrestrial vertebrates are spherical cellular organelles that stain for lipids, have no discernible internal structure, and often contain carotenoids and possibly other chemicals. A survey of 97 species of anuran amphibians (frogs and toads) revealed that all species of 16 families surveyed possessed yellow oildroplets of varying size in the cells of the pigment epithelium, except for three species that appear to have secondarily lost them during evolution. Furthermore, 25 species of six families also possess colorless oildroplets at the distal end of the inner segments of single cones and principal cones of the double-cone system; two species of the Ranidae appear to have secondarily lost such retinal oildroplets. Every species possesses epithelial or retinal oildroplets or both. Lastly, small oildroplet-like inclusions were discovered in the red blood cells of two species. All of Walls' ('42) summary generalizations about anuran oildroplets are incorrect: oildroplets are not restricted to the Ranidae, are not yellow when found in the cones, and do not correlate with photoactic behavior in 87 species. Evidence is reviewed suggesting that the primary function of anuran oildroplets is chemical storage, perhaps related to the visual pigment cycle. Oildroplets in the cones may additionally act as filters of ultraviolet radiation.


Evolutionary processes and evolutionary noise at the molecular level. II. A selectionist model for random fixations in proteins.

On account, notably, of a competition between different component functions for individual sites in polypeptide chains, each protein molecule represents a functional compromise, with some functions optimized, but the overall state of the molecule "suboptimal". The proposal is made that the selection coefficient relating to a protein molecule under given conditions can in principle be broken down into partial selection coefficients relevant to the different functions that the molecule carries out. At general-function sites, each fixation improves some function, while others deteriorate, at first nonsignificantly, and the overall adaptive state of the molecule fluctuates around its maximum. A selective mechanism is described whereby kaleidoscopic changes in primary structure at variable sites are indefinitely promoted, independently of any environmental changes and with the molecule remaining close to a state of maximal overall adaptation. The paradoxical aspect of this proposal is analyzed. The implication of specific functions in substitutions at general-function sites is noted. Further, it is shown that a certain category of changes in the internal environment of the organism can be integrated into the constant-environmental model for selection. Genetic sufficiency is considered a notion more adequate than genetic optimality for describing biological fitness and for providing a basis for the present model. On this basis selection occurs without genetic load. Multipolymorphism is one of the consequences. Several lines of evidence, in particular observations on polymorphism in deep sea organisms, seem to support the model. It is pointed out that it provides a theoretical foundation for a molecular evolutionary clock. The theoretical constancy of the clock depends on the constancy of functional density. The question of the evolution of functional density is examined. Comparisons of observed substitution frequencies with values expected on a random basis are rejected as a measure of the contribution to evolution of nondetermination. They are considered to reflect a hierarchy in the resistance of the molecules to different amino acid residues as substituents. A limited component of "true" randomness, again accompanied by selection, is on the other hand provided by the model. Most amino acid substitutions are considered evolutionary noise, even though noise compatible with selection. It is proposed that evolutionary significant substitutions may be identified by monitoring changes in functional density and weighted functional density.


Empirical relationship between the number of nucleotide substitutions and interspecific identity of amino acid sequences in some proteins.

There are three different methods of estimating the number of nucleotide substitutions between a pair of species from amino acid sequence data, i.e. the Poisson correction method, random evolutionary hit method, and counting the actual but minimum number of nucleotide substitutions. In this paper the relationships among the estimates obtained by these methods are studied empirically. The results obtained indicate that there is a high correlation among these estimates and in practice any of the three methods may be used for constructing evolutionary trees or relating nucleotide substitutions to evolutionary time. The effects of varying rates of nucleotide substition among different sites on the Poisson correction and random evolutionary hit methods are also studied mathematically. It is shown that these two methods are quite insensitive to the variation of the rate of nucleotide substitution.


Gustatory sweating and related syndromes.

Gustatory sweating is a reflex, localized production of sweat associated with sight, smell, or ingestion of food. It involves most commonly the area of the auriculo-temporal nerve where it produces the well known Frey's syndrome. The pathophysiology of the syndrome is described with the aid of diagrams. Anatomically related syndromes are explained, namely the chorda tympani syndrome, crocodile tears, xerophthalmia of the suprageniculate facial nerve palsy, and the partial (otogenic) Horner's syndrome.


Avian arboviruses of the Witless Bay seabird Sanctuary, Newfoundland, Canada.

A virologic and serologic survey of arbovirus infections among seabirds and seabird ticks, Ixodes uriae, on Great Island. Witless Bay, Newfoundland, Canada, was conducted during 1971 and 1972. Kermerovo (Great Island, Bauline) and Sakhalin (Avalon) group viruses previously reported from birds and/or ticks on Great Island were prevalent among avian populations, while conclusive evidence of known nonindigenous serotypes was lacking. Circumstantial evidence-hemagglutination inhibiting antibody-of an unidentified member of the group B tick-borne encephalitis complex transmitted among marine birds of North America by I. uriae is reported. No evidence of human infections with any of these viruses was detected in a small number of biologists doing research on Great Island.


An outbreak of duck virus enteritis (duck plague) in Alberta.

Duck plague (Duck virus enteritis) was disgnosed in a resident population of Muscovy ducks (Cairina moschata) on a small game farm in Alberta. This disease has not been reported previously in Canada. Clinical signs consisted of cyanosis, depression and acute death. Necropsy of two Muscovy ducks revealed lesions typical of the disease. There were ulcerations with pseudomembranes in the small intestine, ulcerations with caseous plaques in the esophagus and eosinophilic intranuclear inclusion bodies in the spleen. Clinical disease with mortality was reproduced in young ducklings injected with tissue homogenates from field cases. All surviving inoculated ducklings seroconverted to highly significant titres of neutralizing antibodies to duck virus enteritis (DVE) virus. All attempts to isolate the agent in embryonating duck eggs or primary tissues cultures of duck and chicken kidney were negative. Identification of the DVE virus was accomplished by serum neutralization with ducklings as the host system.


Nonhuman primate-associated viral hepatitis type A. Serologic evidence of hepatitis A virus infection.

Since 1961, viral hepatitis has been recognized as an occupational hazard among handlers of newly imported chimpanzees and other nonhuman primates. To determine whether previously reported cases were caused by human viral hepatitis type A, we tested paired serum samples from two outbreaks for antibody to hepatitis A antigen (anti-HA) by immune adherence hemagglutination (IAHA), recently available test. In both outbreaks, one of hepatitis transmitted from chimpanzee to man (Michigan, 1964), the second from chimpanzee to chimpanzee, man, and woolly monkey (Connecticut, 1971), serologic data documented recent hepatitis A virus infection among contacts-human and nonhuman primate-of implicated chimpanzees. This confirms serologically a previously noted epidemiologic association between nonhuman primate-associated hepatitis and human viral hepatitis, type A.


Hyperlipidemia due to oxymetholone therapy. Occurrence in a long-term hemodialysis patient.

Marked hypertriglyceridemia and hypercholesterolemia accompanied by angina and a left cerebral thrombosis occurred in a long-term hemodialysis patient following 5 1/2 weeks of oral treatment with oxymetholone, 100 mg/day, a synthetic androgen. After androgen therapy was discontinued, over a three-month period, plasma lipid values progressively decreased below pretreatment values, and clinical symptoms disappeared. During rechallenge with oxymetholone, serum lipid values increased substantially, and the lipoprotein pattern changed from a type IV to a type V. Detailed lipid studies showed subnormal postheparin lipolytic activity and a fast-migrating pre-beta-lipoprotein in a very-low-density lipoproteins (VLDL) fraction. Because of the data linking lipid abnormalities to atherosclerosis and the acceleration of atherosclerosis in long-term hemodialysis patients, great caution should be exercised in administering androgenic steroids to these patients.


Neuromyelitis optica (relapsing type).

A case of relapsing type of neuromyelitis optica following influenza-like illness treated successfully with steroids and corticotrophin has been described. Raised gamma-globulin in CSF was suggestive of an autoimmune disorder. Inspite of six relapses in 8 years she has minimal neurological deficit.


Lipids and lipoproteins of serum and organ tissues in a case of familial hyperlipoproteinemia type II.

Lipids and lipoproteins of serum and various organ tissues of a patient with familial hyperlipoproteinemia homozygous Type II were studied. It was found that low density lipoprotein between densities of 1.006 and 1.063 was a major component of serum of the patient. The isolated low density lipoprotein appeared to be rather similar to that of normal human serum from the viewpoints of chemical composition and physico-chemical properties. Antibody against the low density lipoprotein of the patient reacted not only with the same lipoprotein, but also with normal human low density lipoproteins. Immunoelectrophoresis of the antibody gave a single precipitin line against only the low density lipoprotein. Immunofluorescent techniques using the fluorescein labeled antibody showed localization of low density lipoprotein in cytoplasm of hepatocytes of both the patient and other human. No fluorescences were found in other organ cells except in their capillary tubes and renal glomerulus in the hyperlipoproteinemia Type II. It was noted that tuberous xanthomata contained fairly large amounts of cholesterol and cholesterol ester. Fatty acid composition of neutral lipids and phospholipids from the serum low density lipoprotein, xanthomata, liver and other organ tissues were compared with each other. Chomesterol ester of the xanthomata contained much smaller amounts of polyunsaturated fatty acids as compared with those of the low density lipoprotein and other tissue. Also, composition of free fatty acids in liver differed from those of other tissues.


Serum lipoprotein profile in infectious hepatitis.

There is absence of pre-beta and alpha-lipoproteins in the acute stage of infectious hepatitis. With improvement both clinically and biochemically there is appearance of both lipoproteins, with complete reversal to normal lipoprotein pattern in the end.


Response of nude mice to a mouse hepatitis virus isolated from a wasting nude mouse.

Chronic active hepatitis was produced in nude mice after inoculation with a mouse hepatitis virus from a natural nude case which was incapable of causing fatal infection in haired heterozygous mice without cortisone. Survival time of infected nude mice varied greatly from 1 to 10 weeks after the inoculation. Degeneration and necrosis of hepatocytes, apparition of small and megalocytic hepatocytes, and infiltration of lymphocytes and plasma cells with fibrosis, were most remarkable in those which died 3 weeks or more after inoculation. No such severe hepatic lesions were recognized in heterozygous haired litter-mates after inoculation of the same virus. Necrotic lesions with poor inflammatory reactions were seen in the brain of all infected and dead nude mice, while some cerebral lesions with apparent perivascular infiltration were detected in heterozygous ones only at early stage of infection.


Clostridium perfringens exotoxins. IV. Inhibition of the theta-toxin induced hemolysis by steroids and related compounds.

1. The inhibitory powers of thirty two samples of steroids and their related compounds on the theta-toxin induced hemolysis were assayed. Apparent I50 value of cholesterol, the most potent inhibitor amoung them, was 0.022 muM. 2. Inhibitory powers of sterol acetates were not more than one tenth of those of the corresponding sterol, and steroids without 3-hydroxyl group hardly inhibited the hemolysis. These results suggest that 3-hydroxyl group of sterols plays an important role in the inhibition. 3. The inhibitory power of sterols were strongly affected by steric situation of 3-hydroxyl group for the face of their tetracyclic rings. From these results it is suggested that theta-toxin binds to beta-side of the tetracyclic rings but not to alpha-side. 4. A methylsterol and pentacyclic triterpens with 3-hydroxyl group also inhibited the hemolysis, but steroids and their related compounds either with a hydrophylic or without a hydrophobic group at their ring D had little, if any, inhibitory power.


Cell lines established from (BALB/cx DBA/2)F1 mice which show parent type susceptibility to murine leukemia viruses.

The susceptibility to two tissue culture cell lines established from (BALB/cx DBA/2)F1 (CDF1) mice to murine leukemia viruses (MLVs) was compared with those of various cell lines. While embryo cells of CDF1 mice were resistant to both N- and B- tropic MLVs, one cell line was susceptible to N-tropic MLV and the other to B-tropic MLV. So the cell lines of CDF1 mice had not the same susceptibility to MLVs as CDF1 embryo cells had. Cell lines established from N- and B-type inbred mice had kept their original susceptibility to MLVs. These results suggested the phenotype of some F1 hybrid mouse cells tended to change into either parental type after establishment as cell lines.


Elevated levels of IgE antibodies to ascaris and mite antigens in Papua New Guinea.

Levels of IgE as well as specific IgE antibodies to ascaris and mite have been studied in two groups in Papua New Guinea (PNG), patients with primary liver carcinoma (PLC) and normal blood donors, and in two groups in Japan, normal subjects and asthmatic patients. A radioallergosorbent test was used to measure IgE antibodies to ascaris and mite antigens. Much higher levels of IgE were found in the Papua New Guinean subjects than the normal Japanese subjects. Also levels of IgE antibodies to ascaris were much higher in Papua New Guineans than in the Japanese subjects. Though similar results were observed in IgE antibodies to mite, difference between normal Papua New Guineans and normal Japanese was small, whereas difference between the patients with PLC in PNG and the normal Japanese subjects was very large. Highest levels of IgE antibodies to mite were detected in Japanese with asthma. No correlations were observed among IgE levels, levels of IgE antibodies to ascaris and levels of IgE antibodies to mite. It is concluded that both ascaris and mite IgE antibodies contribute the very high levels of IgE seen in Papua New Guineans, but many other factors may be operative.


Effects of cytochalasin B and dibutyryl cyclic AMP on intraleukocytic bactericidal activity.

Cytochalasin B (CB) is known to regulate the movement of intracellular microfilaments system. In this experiment, the effect of CB on the intraleukocytic bactericidal activity was first studied and concluded that CB inhibited the intracellular bactericidal activity remarkably. Addition of CB resulted in increased nitroblue tetrazolium (NBT)-dye reduction, while the specific activities of glucose-6-phosphate dehydrogenase (G-6-PD) and reduced nicotinamide-adenine dinucleotide phosphate (NADPH) oxidase of CB-treated neutrophils exhibited within the normal range. But the uptake of Glucose-1-14C or -6-14C was markedly inhibited by CB treatment. However, the following substances inhibited NBT-dye reduction in decreasing order: cyclic adenosine 3', 5'-monophosphate (cAMP), N6-O2' dibutyryl cyclic adenosine 3', 5'-monophosphate (DBcAMP), prostaglandin F2alpha (PGF2alpha) and prostaglandin E1 (PGE1). These drugs also decreased hexose monophosphate shunt (HMS) enzyme activities. Addition of CB and DBcAMP decreased the intracellular bactericidal activity of neutrophils. But CB had no effect on intracellular levels of cAMP. From the results obtained, it is likely that intracellular bactericidal phenomena of human neutrophils are controlled by cyclic AMP cascades and by the microfilaments system, separately.


Effects of various agents on synaptosomal adenylate cyclase activity in the absence and presence of the boiled supernatant.

A study was made of the effects of various agents on adenylate cyclase in synaptosomes in both the absence and presence of the boiled supernatant from rat cerebral cortex. The activity of adenylate cyclase in these preparations was inhibited by the sulfhydryl reactive agent p-chloromercuribenzoate. Sulfhydryl compounds such cysteine, glutathione and Coenzyme A stimulated the enzymic activity in both the absence and presence of the boiled supernatant. The chelating agent 1.2-bis-(2-dicarboxymethylaminoethoxy)ethane caused a stimulation of the enzymic activity with and without the presence of the boiled supernatant. Adenine nucleotide (adenine, adenosine, AMP and ADP), GTP, Pi and carbamylcholine seemed to have little effect. The stimulatory substance in the boiled supernatant was estimated to have a molecular weight in the range of 1,000-1,300.


Japanese encephalitis surveillance in China (Province of Taiwan) during 1968-1971. I. Geographical and seasonal features of case outbreaks.

During 1968-1971 Japanese encephalitis (JE) surveillance was conducted with WHO-assisted programs in Taiwan area. Emphasis was placed on: (1) active case-finding by hospital visits; (2) blood collection from every patient at the suitable time; and (3) the enforcement of standard diagnostic criteria on the results of hemagglutination-inhibition tests. Each year, approximately 90% of reported patients were etiologically examined. JE virus etiology was established in 277 (1968), 279 (1969), 269 (1970) and 158 (1971) cases. Despite the apparent concentration of cases in several cities, practically all the cultivated plains and basins were established as potentially endemic for JE. JE outbreak is found to have been clearly associated with season, not only in subtropical but also in tropical Taiwan. Each year it occurred consistently in mid-July in the southernmost county, 2-3 weeks after the peak of rice transplantation and its duration was a few weeks. On the western side of the island, although less consistently, JE outbreak seems to occur one to three weeks later than in the south. Outbreak in eastern Taiwan was a mid-September phenomenon for 3 years. Observed seasonal lag, which was significant between two neighboring localities on same latitude, suggests that paddy water management is more directly related to date of outbreak than local climate. Seasonal characteristics further suggests that the area may be separated into eight ecologic subdivisions. JE-virus transmission to a man during April at temperatures below the average of 20 C was discussed.


Japanese encephalitis surveillance in China (Province of Taiwan) during 1968-1971. II. Age-secific incidence in connection with Japanese encephalitis vaccination program.

A baseline age/morbidity pattern for Japanese encephalitis (JE) epidemic in China (Province of Taiwan) in the 1960s was sought in JE surveillance. The pattern is characterized by: (1) high incidence in age groups 2-4 years, (2) moderately high incidence in age groups 5-7 years, (3) negligible incidence in children under 1 year of age as well as in adults, and (4) the variability of relative height of peak incidence co-incident with the JE-epidemic size in situ. Concurrently with surveillance, a JE-vaccination program was implemented by the Government. The children of target age groups received two doses of vaccine each during April. Vigilant supervision was given to the campaign to achieve a high coverage rate with two-dose vaccination and meticulous record-keeping. During 1969-1972, 2-year olds were the exclusive target; 52.2% (1969), 71.5% (1970), 74.5% (1971), and 83.0% (1972) of the target age group were covered. A considerable change from the baseline age/morbidity pattern seen in 1968 occurred during the above period. The comparison of epidemicity-adjusted incidence by year indications that incidence reduction in the 2- and 3-year old children in 1971 were substantial and suggests the possible impact of vaccination. The total JE incidence for the period 1968-1971 appears to be much lower in vaccinated population (23 reported and 8 confirmed/0.9 million) than in unvaccinated population (1,330 reported and 679 confirmed/2.2 million).


Evolution  of the digestive microflora in a unit  of specified-pathogen-free mice: efficiency of the barrier.

RhoIco strain], barrier-maintained since 1970, is described. Some "contaminants" appeared spontaneously during the period 1971-1973, but microorganisms belonging to the genera Lactobacillus Streptococcus, Enterobacteria, Escherichia and Bacillus remained stable. The methods of investigation used were not suitable for the assessment of strictly anaerobic strains. The stability of the digestive microflora durine the last 2 years of this study is believed to be related to the skill and conscientiousness of the technicians responsible for the daily care of these mice.


Renomedullary interstitial cells in the spontaneously hypertensive rat.

Renomedullary interstitial cells (RIC) are known to synthesize and release prostaglandins which may play a significant role in the development or severity of hypertension. The medulla of the spontaneously hypertensive rat contains RIC which are morphologically very similar to those previously described in the normotensive rat. The granularity of the RIC, however, was increased in the spontaneously hypertensive rat compared to normotensive Wistars (9.6 +/- 2.34 versus 5.3 +/- 2.05 granules per cell, respectively, p less than 0.001) or treated spontaneously hypertensive rats (7.2 +/- 1.65 granules per cell, p less than 0.001). Granule counts also increased in the presence of mild and moderate degrees of renal arteriolar sclerosis, but decreased in long standing hypertension with more severe and extensive lesions involving both arteries and arterioles. These results are consistent with the hypothesis that the RIC respond to an elevation of blood pressure in the spontaneously hypertensive rats by increased release of antihypertensive substances. In addition, the decrease in granularity of the RIC in the presence of extensive renal arteriolar and arterial damage suggests reduced ability to compensate for the elevated blood pressure and thus may contribute to the acceleration of hypertension.


Uptake of circulating iron by the duodenum of normal mice and mice with altered iron stores, including sex-linked anemia: high resolution radioautographic study.

Using electron microscopy radioautography, the deposition of intravenously administered iron in the duodenal epithelium was studied in normal mice, iron-overloaded and iron-deficient mice, and in mice with X-linked anemia (gene symbol sla) 4 and 24 hours after injection of 59Fe. The resolution of radioautography with 59Fe was determined with a line source and the distance from the hot line within which half of the grains fell (HD value) was 1650 A. In normal, iron-overloaded, and sla mice, radioiron was localized in the undifferentiated crypt cells at 4 hours and in the absorptive cells of the luminal half of the villi, at 24 hours. At both times, the vast majority of the grains was seen over the areas rich in free ribosomes and rough endoplasmic reticulum. In iron-deficient mice, grains were not found at either time. The amount of iron incorporated in the crypt cells was related to the size of the body iron stores. It is postulated that the amount of iron incorporated in the crypt cells is the result of interaction between uptake and return to the blood of the circulating iron. Once the crypt cells have differentiated into absorptive cells, the uptake and recirculation of iron from and to the blood would cease, leaving an amount of "message" iron which determines the absorptive cell's subsequent capacity for iron transfer to the plasma. In sla mice, in spite of tissue iron deficiency, the amount of iron deposited was similar to that of normal mice and markedly increased after treatment of the anemia. The accumulation of iron in the crypt cells may result from the decreased return to the blood of the incorporated circulating iron, due to the postulated deficient iron carrier mechanism in sla, or may be a consequence of increased avidity of the absorptive cells for circulating iron. In either event, the result would be that the absorptive cell receives an inappropriate message with resulting inappropriate absorption of iron.


The existance of a group translocation transport mechanism in animal cells: uptake of the ribose moiety of inosine.

After exposure to inosine, transport-competent plasma membrane vesicles isolated from SV-40-transformed Bal/c 3T3 cells accumulate intravesicular ribose 1-PO4 at a concentration 200-fold greater than the extravesicular concentration. An analysis of the purine nucleoside phosphorylase activity distribution in various subcellular fractions, relative to other enzyme activities, indicated the presence of plasma membrane-associated purine nucleoside phosphorylase activity. The plasma membrane vesicles appear relatively impermeable to hypoxanthine. However, hypoxanthine, which is a competitive inhibitor of the transport reaction, is the only compound tested capable of mediating efflux of already accumulated ribose 1-PO4. In addition, hypoxanthine does not result in the efflux of transported uridine which is accumulated in these membrane vesicles as uridine. Exogenous ribose 1-PO4 neither results in counterflow nor does it inhibit the original uptake reaction. The following transport reaction is proposed: uptake occurs by group translocation, mediated by membrane-localized purine nucleoside phosphorylase. The data are consistent with sites for inosine and hypoxanthine being on the outer membrane surface whereas the ribose 1-PO4 site is only on the inner surface.


Transport of amino acids in intact 3T3 and SV3T3 cells. Binding activity for leucine in membrane preparations of ehrlich ascites tumor cells.

Transport of amino acids into 3T3 and SV3T3 (SV40 virus-transformed 3T3) cells was measured on glass cover slips. The 3T3 and SV3T3 cells contain both A (alanine preferring) and L (leucine prefferring) systems for neutral amino acid transport. Initial rates of uptake of amino acids are about twofold higher in SV3T3 than in 3T3 cells. Other parameters measured, however, do not indicate marked differences in the transport of amino acids by the two cell types. L-system amino acids, such as leucine, are subject to trans-stimulation in both cell lines, whereas A-system amino acids, such as alanine and glycine, are not. Leucine was transported to higher levels in confluent cells than in nonconfluent cells. Glycine, however, shows distinctly less transport activity as the cells become confluent. Ehrlich ascites cell plasma membranes were prepared and assayed for amino acid-binding activity. Leucine-binding activity was detected by equilibrium dialysis in Triton X-100-treated membrane preparations.


Effect of essential fatty acid deficiency on activity of liver plasma membrane enzymes in the rat.

Liver plasma membranes (LPM) were isolated from rats fed an essential fatty acid-supplemented diet (+EFA) or from rats fed an essential fatty acid-deficient diet (-EFA). The proportions of linoleate and arachidonate in membrane total fatty acids in the -EFA preparations were one-half or less than the values for the +EFA preparations. Basal, F-, or glucagon-stimulated adenylate cyclase activities were significantly lower in EFA-deficient livers than in nondeficient ones. Addition of GTP significantly enhanced glucagon-stimulated adrenylate cyclase in both groups, but extent of stimulation above basal was greater in EFA-deficient livers. Portal vein injection of glucagon in vivo resulted in significantly higher cAMP formation in +EFA livers than in -EFA livers. When glucagon was used in vitro at 1-1,000 nM, stimulation of adenylate cyclase remained lower in EFA-deficient membranes, but extent of stimulation above basal activity was larger in -EFA membranes than in +EFA. Total Na+, K+ (Mg2+)-ATPase from EFA-depleted LPM exhibited significantly higher values of apparent Km and Vmax-5'-Nucleotidase activity, in contrast, was considerably decreased in EFA-deficient rats. These findings show that, in animals, changes in unsaturated fatty acid composition can affect the properties of membrane-bound enzymes. These alterations could be due to changes in membrane physical properties and/or prostaglandin formation.


Avian tumor virus interactions with chicken fibroblast plasma membranes.

A method is described which will rapidly measure the binding of avian tumor viruses (ATV) to plasma membrane receptors. With this procedure it may be shown that Rous sarcoma virus pseudotypes bind to protease-labile, heat-stable structures on the surface of chicken embryo fibroblast (CEF) plasma membranes. The binding sites for ATV subgroups A and B appear distinct, and membranes from genetically resistant CEF bind as well those of sensitive CEF.