Involvement of cholecalciferol metabolism in birds in the adaptation of calcium absorption to the needs during reproduction. 1. The metabolism of calcium and cholecalciferol in quail (Coturnix coturnix japonica) and chicken (Gallus domesticus) during maturation was correlated to gonadal activity and plasma oestrogen levels. 2. Birds with undeveloped ovaries (immature), developed ovaries but not laying (mature), and after laying 3-8 eggs (laying), were used in the first series. 3. Birds in which egg production had been arrested by Nicarbazin, were used in the second series. 4. Plasma 17 beta-oestradiol and calcium were elevated in the mature bird, with no further change in the laying bird. Kidney 25-hydroxycholecalciferol-1-hydroxylase and intestinal calcium-binding protein increased slightly in the mature bird, whereas they were grossly elevated in the laying bird. 5. Calcium and phosphorus absorption were markedly elevated in the laying bird. 6. No changes were noted in plasma 25-hydroxycholecalciferol, at any stage of maturation. 7. During the arrest of egg production by Nicarbazin, 17 beta-oestradiol level, calcium concentration of plasma, and medullary bone were maintained. Kidney 25-hydroxy-cholecalciferol-1-hydroxylase, intestinal calcium-binding protein and absorption of calcium were strikingly reduced. 8. The results suggest that changes in calcium absorption and cholecalciferol metabolism during maturation in birds are not directly affected by gonadal hormones; they appear to represent an adaptation to the increased calcium needs due to medullary bone formation and, more importantly, to the large losses of calcium imposed by shell formation.


A study of bovid herpesvirus 2 infection in calves inoculated intradermally or intranasally. Bovid herpesvirus 2 infection was studied in calves exposed to the virus by intradermal inoculation of the skin of the left cheek or by nasal spray. In either case a localised infection developed and virus replication was shown to occur mostly in the tissues of its primary localisation, i.e. the skin of the left cheek or the nasal mucosa. There were neither secondary lesions, except at the site of virus injection, nor any serious systemic involvement on the part of the animals. The virus was also recovered from some areas of the skin (right cheek, perineum and scrotum) that were free of macroscopic lesions; moreover, intranuclear inclusions were found in several tissues of the nervous system (brain, superior cervical, stellate and Gasserian ganglia) which did not show any signs of inflammatory or degenerative changes. These findings suggest that the skin and the nervous system play an important role in the naturally-occurring disease since they could be the sites where the virus is maintained latently in the host.


A new freeze-dried living virus vaccine against sheep-pox. A sheep-pox virus strain has been adapted and multiplied in primary lamb kidney cell cultures. The main characteristics of the strain have been verified in vitro after clones were isolated, and the results confirmed its identity. The safety and the potency of the strain have been investigated in sheep. The inoculation of the strain to sheep was followed by a post-vaccinal reaction materialised by a nodule at the site of inoculation and an increase of temperature by about 1 degree C. No reactions adversely affecting pregnancy have been noted. Immunisation was demonstrated by an increase in the level of neutralising serous antibodies and protection against the pathogenic virus. The immunity tended to decrease during the second year after primovaccination and a yearly booster vaccination appeared to be necessary. Primovaccination of lambs over 2 months of age produced a better immunity, especially when the lambs were born from vaccinated ewes. This strain forms the active principle of a freeze-dried vaccine containing no adjuvant of the immunity.


Immunisation against panleukopenia: early development of immunity. The time necessary to obtain the immunity of cats against Panleukopenia has been studied by means of a modified live vaccine. This vaccine makes it possible to obtain a very early post-vaccinal immunity: the full immunity is reached 72 hr after the inoculation of the vaccine by the subcutaneous route. Furthermore, we have demonstrated that a sensitive kitten can be admitted in a contaminated environment immediately after vaccination without showing any clinical evidence of the disease.


Bovine adenoviruses--IV. Two mixed antigens for routine serodiagnosis by complement fixation reaction. The frequency of bovine adenovirus infections occurring in cattle has been grossly underestimated up to the present day. Primary isolation of serotypes belonging to bovine subgroup II is cumbersome. Hardly any laboratory has used all 8 officially-recognised bovine serotypes in seroneutralisation-tests, which should be done when this type-specific method is used for serodiagnosis. The complement fixation test, known as group-reactive from human adenovirus serology, has failed to disclose the true incidence of bovine infections, as until recently the importance of a novel additional group-reactive bovine adenovirus antigen has been undisclosed. Here we describe production, composition and performance of two mixed antigens for complement fixation reactions, which take into account recent findings by our team on peculiarities of bovine adenovirus antigens. Mixed antigen 1 contains bovine serotypes 1, 2 and 3 and detects group-specific antibodies of the classical mastadenoviruses. Mixed antigen 2 contains bovine serotypes 4, 5, 6, 7 and 8 and determines group-specific antibodies against the novel paramastadenoviruses. In addition, each antigen is capable of demonstrating complement-fixing type-specific antibodies in sera against the respective types included in each mixed antigen, with a net enhancing effect produced by mixing. Use of the 2 mixed antigens promptly shows whether bovine adenoviruses, presently recognised as well as unclassified, are involved in a given outbreak of respiratory or enteric disease of cattle. If needed, the responsible type may be determined in a second step of serological investigation.


Carbohydrate composition of serum low and high density lipoproteins of nonhuman primate species. 1. Carbohydrate composition of serum low and high density lipoproteins obtained from 5 nonhuman primate species (chimpanzee, patas, baboon, rhesus, and spider) and humans was studied. 2. Individual lipoproteins were isolated from pooled sera of each species by ultracentrifugal flotation between the densities 1.019-1.063 for LDL-2; 1.063-1.12 for HDL-2; and 1.12-1.21 for HDL-3. After delipidation, sialic acid, fucose, glucosamine, mannose, galactose, and glucose were determined on apo LDL-2, apo HDL-2, and apo HDL-3. 3. Glucosamine, galactose, and mannose constituted a major component of the sugars in apo LDL-2, with similar relative proportions in all species. Sialic acid, fucose, and glucose formed a minor component, the proportions of which varied greatly among the species. 4. Unlike apo LDL-2, sialic acid, fucose, and glucosamine constituted the bulk of the sugars in apo HDL-2 and apo HDL-3. Mannose, galactose, and glucose were minor components, with galactose predominating. 5. Qualitative differences were observed in electrophoretic mobilities of apo HDL-2 and apo HDL-3 on polyacrylamide gel. One faster moving band was unique to chimpanzee. 6. Intraspecies differences in the content of sialic acid and fucose of apolipoproteins may be related to lipoprotein metabolism and species susceptibility (or resistance) to either spontaneous or diet-induced atherosclerosis.


A comparative study of some more important experimental animal peroxide metabolism enzymes. 1. We have studied and compared the peroxide metabolism enzymes (SOD, P and C) of the main organs of fresh-water mollusc, chicken, mouse, guinea-pig, rabbit, cat and dog. 2. The liver exhibited the highest SOD activity. The enzymatic activities of the organ homogenates of the guinea-pig stand out in comparison with the values for the other homogenates examined. 3. The liver, kidney and total brain homogenates of the chicken and the vertebrates display no, or only a very low P activity. The highest P activities were measured in the haemolysates. 4. The C and SOD values exhibit a certain parallelism. 5. The peroxide metabolism enzyme activities calculated by utilizing the protein measurements permit the establishment of a more realistic enzymatic activity.


Enzymes of galactose utilization in the rat tapeworm, Hymenolepis diminuta. 1. Crude enzyme preparations from Hymenolepis diminuta contained galactokinase, galactose 1-phosphate uridyl transferase and UDPgalactose 4-epimerase activity, although their specific activities were low. 2. Galactose 1-phosphate non-competitively inhibited galactose phosphorylation. This inhibition, together with the low specific activities of the enzymes in the pathway of galactose utilization, probably accounts for the inadequacy of galactose as a main nutritive carbohydrate for development of the worm.


Differential lipid control of (Na+ + K+)-ATPase in homeotherms and poikilotherms. 1. (Na+ + K+)-ATPase from homeotherms and poikilotherms demonstrate non-linear thermal dependence for ATP hydrolysis. Apparent energies of activation from crab nerve preparations are less than those of brain or kidney preparations from beef, rabbit, sheep or ground squirrel. 2. Crab nerve (Na+ + K+)-ATPase is less sensitive to inhibition by ouabain than that from beef or ground squirrel; lower rates of [3H]-ouabain binding and reduced amount of drug bound at equilibrium are found. 3. K+-activated acyl-phosphatase is similar in all preparations. 4. Fluorescence polarization of 12-AS labelled membranes demonstrate greater mobility of crab nerve lipids compared to beef brain which has a thermal transition at 20-25 degrees C. Crab nerve is linear in this range.


Glucose phosphorylation and dephosphorylation in chicken liver. 1. Glucokinase was absent from chicken liver and only the low Km hexokinases, inhibited by AMP, ADP but not ATP, were present. 2. The Km of chicken liver glucose-6-phosphatase for glucose-6-phosphate was reduced from 5.65 to 3.75 mM following starvation, and the enzyme was inhibited by glucose. 3. Starvation of chickens for 24 hr slightly lowered the hexokinase activity and doubled glucose-6-phosphatase activity; it did not change subcellular distribution of the enzymes. Oral glucose rapidly restored the activities to fed values. 4. It was concluded that glucose uptake into, and efflux from, chicken hepatocytes, was regulated by the activity and kinetic characteristics of glucose-6-phosphatase and by the glucose-6-phosphate concentration, and that the hexokinases had little regulatory function.


Regulation of carbohydrate metabolism in mouse liver. Effect of glucagon on gluconeogenic/glycolytic flux in isolated perfused livers. 1. Glucagon stimulated gluconeogenesis from both [U-14C]lactate and [14C]xylitol in isolated perfused mouse liver. 2. Addition of cyclic AMP also stimulated gluconeogenesis from [U-14C]lactate. 3. Glucagon caused a rapid (2.5 min) 12-fold increase in hepatic cyclic AMP but not cyclic GMP concentration. 4. Glucagon caused a rapid and stable decrease in hepatic fructose 1,6-diphosphatase activity measured in vitro. 5. The results are interpreted to indicate that glucagon stimulates hepatic gluconeogenesis in mice via cyclic AMP by two different mechanisms: (a) increased substrate uptake (i.e. utilization) and (b) increased gluconeogenic efficiency (i.e. inhibition of alternate substrate fates).


The effects of ACTH on adrenal steroidogenesis and blood corticosteroid levels in the echidna (Tachyglossus aculeatus). 1. The effects of short-term (S.T., 30 min) and long-term (L.T., 4 days) administration of ACTH on peripheral blood corticosteroid levels and on in vitro steroidogenesis were investigated. 2. Control levels of cortisol, corticosterone and aldosterone were 58 +/- 12, 130 +/- 26 and 10 +/- 6 (SEM) ng/100 ml respectively. 3. Corticosterone was 70% higher after S.T. and 150% higher after L.T., when cortisol was 800% higher. 4. Adrenal homogenates from control echidnas converted [14C]progesterone predominantly to 11-deoxycorticosterone (45%) and 11-deoxycortisol (12%). 5. After L.T. the principal product was corticosterone (25%), but S.T. had no effect. 6. In control echidnas the Km and V for 11 beta-hydroxylation of 11-deoxycorticosterone were 20 microM and 2.8 rho mol/min/mg respectively. After L.T. V increased to 10 rho mol/min/mg.


Biochemical studies of taste sensation--VIII. Partial characterization of alanine-binding taste receptor sites of catfish Ictalurus punctatus using mercurials, sulfhydryl reagents, trypsin and phospholipase C. 1. Taste receptors for L-alanine in the channel catfish Ictalurus punctatus have been partially characterized. The binding activity, which is localized to a sedimentable fraction (Fraction P2), was assayed with L-[3H]alanine as the ligand. 2. Addition of HgCl2 or p-mercuribenzoate to the assay at 0.1-1 mM markedly inhibited binding. The effect was not reversible and was unaffected by increased L-alanine in the binding assay. 3. The sulfhydryl reagents iodoacetate, 5,5'-dithiobis(2-nitrobenzoic acid), arsenite, and N-ethylmaleimide did not show appreciable inhibition of binding. The results suggest that the inhibitory effect of mercurials is not on specific sulfhydryl groups at alanine-binding sites. 4. Treatment of Fraction P2 with phospholipase C decreased binding activity and treatment with trypsin led to increased binding activity.


Zinc effects on LDH, MDH and alkaline phosphatase from cultures of fathead minnow cells. 1. Three purported zinc metalloenzymes have been investigated from cell cultures of the fathead minnow (Pimephales promelas). 2. With the addition of increasingly higher concentrations of zinc to the tissue culture medium, the specific activity of LDH increased. 3. The results with MDH were equivocal. 4. The specific activity of alkaline phosphatase decreased in the presence of increasing amounts of zinc in the growth medium. 5. Zinc exogenously added to the LDH enzyme assay did not alter the LDH enzyme activity of cells grown without zinc.


Lipid and carbohydrate metabolism in euthyroid and hypothyroid chick embryo (Gallus domesticus). 1. The effect of propylthiouracil (PTU)-induced hypothyroidism on carbohydrate and lipid metabolism was studied in the chick embryo. 2. A single dose of PTU (250 micrograms/embryo) was administered on day 11 and embryos sacrificed on day 20 of incubation. 3. Thyroid glands were significantly enlarged (6 fold) by PTU administration. 4. Increased thyroid weight was associated with growth retardation and decreased plasma thyroxine levels. 5. Plasma glucose level was lower and phospholipids were significantly higher in the hypothyroid embryo. 6. Liver lipid concentrations in the control and hypothyroid embryos were not different but were significantly higher in both groups when compared to previously reported values in the young chick. 7. In contrast to PTU treatment after hatching, liver glycogen levels were not increased in the hypothyroid chick embryo. This was attributed to the high lipid nutrient condition of the chick embryo since a high lipid diet in the young chick decreased hepatic glycogen accumulation significantly.


Ascorbic acid-dependent collagen formation in penaeid shrimp. 1. This study tested the hypothesis that black death, the ascorbic acid (AsA) related disease of penaeid shrimp, is related to collagen underhydroxylation. 2. Collagen measured as hydroxyproline (HYP) in healthy Penaeus californiensis (Holmes) and P. stylirostris (Stimpson) of a wide range of masses were determined. The results revealed a logarithmic relationship between total body collagen HYP and body weight fitting the equation y = 90x1.18 where y = total collagenous HYP (microgram) and x = body weight (g). 3. Shrimp tissues most subject to mechanical trauma (subcutis, hindgut and gills) had the highest collagenous HYP levels and were most consistently and severely affected by an ascorbic acid (AsA) deficiency disease. 4. Prolyl hydroxylase (PH) activity was demonstrated in tissues of P. californiensis and P. stylirostris by hydroxylation of [3,4-3H]proline. 5. AsA was required for shrimp PH activity using a chicken embryo substrate. 6. Nutritional trials revealed that dietary AsA was required for proline hydroxylation in collagen formation in P. californiensis.


Effects of 5-hydroxytryptophan on fragmentation of REM sleep in alcoholics. The authors studied the effects of oral loading with 5-HTP on REM fragmentation in a group of alcoholics who were abstinent following acute ethanol withdrawal. Decreased fragmentation was found in the majority of subjects, and those subjects with low baseline REM efficiency (i.e., greater fragmentation) showed more improvement from the drug than did subjects who were less impaired initially. The authors suggest that there is an organic decrement of serotonin during ethanol withdrawal which is partially reversed by 5-HTP loading.


Determination of adenosine 3',5'-cyclic monophosphate levels in tissues of the free living nematode, Panagrellus redivivus. 1. Tissue levels of adenosine 3', 5'-cyclic monophosphate (c-AMP) were determined by a protein binding assay in mixed populations of the free living nematode Panagrellus redivivus. 2. The values were 2.6 pmoles/mg dry weight (range 1.3-3.4 pmoles). 3. The identity of the c-AMP assayed was confirmed by both anion exchange and TLC as well as enzymatic degradation.


Seasonal variations in the renal cortical (Na+ + K+)-ATPase and Mg2+-ATPase of a hibernator, the ground squirrel (Spermophilus richardsonii). 1. The specific activity of renal cortical (Na+ + K+)-ATPase of the Richardson ground squirrel is markedly reduced during hibernation, in contrast to the specific activity of the accompanying Mg2+-ATPase which is markedly increased. 2. The sensitivity of (Na+ + K+)-ATPase to inhibition by ouabain is unchanged by hibernation. 3. Both the non-linear thermal dependence of (Na+ + K+)-ATPase and the linear thermal dependence of Mg2+-ATPase are also unchanged by hibernation. 4. The energy of activation of both enzymes is unchanged during hibernation, or by comparison with that determined in awake controls. 5. There is no evidence for inherent "cold resistance" in these enzyme preparations compared to similar preparations from the non-hibernating rabbit. This parameter does not change during hibernation. 6. Both the rate and amount of specific [3H]-ouabain binding to the renal cortical preparations of (Na+ + K+)-ATPase decrease during hibernation. This decrease matches the fall in enzyme activity so that the ratio of pumping sites/unit of enzyme activity shows no seasonal variations. 7. These findings suggest that the amount of renal cortical (Na+ + K+)-ATPase enzyme falls during hibernation, but that the enzyme which remains functions with the same thermodynamic efficiency and identical biochemical characteristics of that found in the awake summer controls.


Difference in renal alpha-galactosidase levels in male and female Chinese hamsters (Cricetulus griseus). 1. The activity of alpha-galactosidase was found to be significantly higher in the kidney of female than that of male Chinese hamsters in a highly inbred colony but its activity in liver, heart and spleen remained similar between female and male animals. 2. Partially purified renal alpha-galactosidase by sequential column chromatography on Sepharose 6B and DEAE-Sepharose CL-6B showed identical elution profiles, pH optima (4.5), KmS (4.4 mM) and heat-inactivation curves between enzymes of male and female animals. 3. Thus, the observed higher activity of renal alpha-galactosidase in the females was due to elevated enzyme concentration, not a result of enzyme polymorphism.


NAD glycohydrolase activity in dog cardiac tissue. 1. Dog heart tissue suspension hydrolyzes NAD, NADP and NMN, and releases nicotinamide stoichiometrically. 2. Maximum activity was observed at 50 degrees C and the activation energy was 10 kcal/mol. 3. Optimum pH range was 6.2-7.6. 4. Compounds with adenine-ribose moiety increased the enzymatic activity. 5. Nicotinamide released during incubation produced reaction nonlinearity. 6. Km for NAD and NADP were about the same; Vmax was higher for NAD. Similar findings have been reported for rabbit heart. 7. Dog enzyme appears to be more sensitive than the rabbit enzyme to noncompetitive inhibitors. 8. Pyrophosphatase activity was not detected in dog heart in contrast to rabbit and rat heart preparations.


Determination of guanosine 3',5'-cyclic-monophosphate in tissues of the free living nematode, Panagrellus redivivus. 1. Tissue levels of guanosine 3',5'-cyclic monophosphate (c-GMP) were determined by radioimmunoassay in mixed-age populations of the free living nematode Panagrellus redivivus. 2. Cyclic-GMP was identified by chromatography (ion exchange and thin-layer) and by enzymatic degradation.


Characterization of mevalonate-activating enzymes in the neonatal chick liver. 1. The 5-phosphomevalonate (MVAP) and 5-pyrophosphomevalonate (MVAPP) formation by cell-free extracts from 7-10 days chick liver shows an absolute nucleotide requirement, ATP being the most effective phosphate donor, though ITP and UTP can be used less effectively. 2. Mn2+ is a better activator than Mg2+ at low concentrations (0.1-5.0 mM). At higher concentrations (10.0 mM) Mn2+ produces a clear decrease in the MVAP formation, whereas the maximum MVAPP formation occurs in the presence of 10.0 mM Mg2+. 3. Mevalonate-activating enzymes maintain their activities for 48 hr at 4 degrees C and 24 hr at 37 degrees C. No MVAP is formed when the extracts are heated to 65 degrees C for 10 min. 4. Unlike other vertebrate mevalonate and phosphomevalonate kinases, these enzymes from chick liver are not activated by -SH group protectors as dithiothreitol, reduced glutathione, cysteine or beta-mercaptoethanol. However, the enzymes are found to be sensitive to thiol binding reagents p-hydroxymercuribenzoate and 5,5'-dithiobis-(2-nitrobenzoic acid).


Enzymes involved in adenine nucleotide metabolism of developing chick embryo liver. 1. This paper describes the changes in the activity of adenylate deaminase, adenylate and inosinate phosphatase, and adenosine deaminase in the developing chick embryo liver. 2. The adenylate and inosinate phosphatase and adenosine deaminase activity appears considerably higher in chick embryo liver with respect to other chick embryo tissues previously examined. 3. During development the control exerted by ATP on AMP breakdown undergoes variations. Consequently, in the first period of incubation AMP is degraded by the direct pathway (AMP-IMP) and in the last period of incubation by the indirect pathway (AMP-adenosine). In the intermediate period (from the 12th to the 15th day of incubation) both pathways may be followed. 4. The ability to synthesize purine nucleotides through "salvage pathway" seems to be acquired by embryonic liver at least at the 15th day.


Relationship between essential fatty acid requirements of aquatic animals and the capacity for bioconversion of linolenic acid to highly unsaturated fatty acids. 1. [1-14C]linolenic acid was injected into the rainbow trout, Salmo gairdnerii, ayu, Plecoglossus altivelis, eel, Anguilla japonica, red sea bream, Chrysophrys major, rockfish, Sebastiscus marmoratus, globefish, Fugu rubripes rubripes and prawn, Penaeus japonicus (molting stage D"1-D2), and the bioconversion of linolenic acid (18:3 omega 3) to highly unsaturated fatty acids such as eicosapentaenoic (20:5 omega 3) and docosahexaenoic (22:6 omega 3) acids was investigated. 2. Linolenic acid was converted to 20:5 omega 3 and 22:6 omega 3 intensively in the rainbow trout, moderately in the ayu, eel and prawn, but slightly in the red sea bream, rockfish and globefish. 3. These results were discussed in relation to the essential fatty acid requirements of the aquatic animals.


Isolation and characterization of a larval lipoprotein from the hemolymph of Manduca sexta. 1. The larval high density lipoprotein (HDL) from the hemolymph of Manduca sexta, isolated by density gradient centrifugation, contains 61% protein, 37% lipid and 2% carbohydrate. 2. The molecular weight of HDL is 6 x 10(5), with two apoproteins of 2.85 x 10(5) and 8.1 x 10(4) daltons. 3. The large apoprotein is destroyed by trypsin treatment of the particle, while the small one is not. 4. Calculations based upon size and composition show that this particle is very different in structure from mammalian lipoproteins. It is proposed that a portion of the apoprotein occupies the central core region.


Red cell uridine monophosphate kinase: a comparative study in various vertebrate species. 1. Starch gel electrophoretic patterns and activity of red cell UMP-kinase were compared in different species of vertebrates. 2. A characteristic electrophoretic pattern for each species of animal studied was found. 3. The mammals differed markedly from the rest of the vertebrate compared in having a faster electrophoretic migration. 4. Significant species differences in total UMPkinase activity could be observed.


Subcellular localization of glucose-6-phosphatase in animal tissues. 1. Glucose-6-phosphatase (EC 3.1.3.9 D-glucose-6-phosphate phosphohydrolase) was found to be localized mainly in the endoplasmic reticulum (microsomal fraction) of all species of vertebrate liver tissue examined. 2. Hepatopancreas tissue from gastropod molluscs was found to be unique in showing the localization of glucose-6-phosphatase in the cytosol (soluble fraction).


Mechanism of L-serine oxidation in Entamoeba histolytica. 1. The enzymatic mechanism of oxygen uptake elicited by L-serine in axenically cultivated trophozoites of Entamoeba histolytica was investigated. 2. Of 22 amino acids examined, only L-serine stimulated oxygen consumption by intact and disrupted amoebae. 3. Pyruvate, a product of serine metabolism, also stimulated oxygen consumption in the amoebae. 4. Characterization of the oxygen uptake elicited by both L-serine and pyruvate, and analysis of the products of L-serine metabolism indicate that the amino acid is first converted to pyruvate. 5. L-Serine dehydratase, which catalyzes the deamination of serine to pyruvate, was detected primarily in the soluble fraction of the amoebae. D-Serine potently inhibited the enzyme, as well as oxygen uptake in the presence of L-serine but not in the presence of pyruvate. 6. The pyruvate formed is oxidized, at least in part, by a novel pyruvate oxidase involving the uptake of molecular oxygen.


Levels of true thymidine kinase and nucleoside phosphotransferase in two strains of Tetrahymena pyriformis under different growth conditions. Activities of typical thymidine kinase and nucleoside phosphotransferase are both present in logarithmically growing tetrahymena pyriformis, GL-1 and ST strains, contrary to previous reports. 2. Activities of thymidine kinase and nucleoside phosphotransferase are also found in both GL-1 and ST strains grown in the defined medium, PPL medium and Neff's medium. 3. The specific activities of both enzymes are very much influenced by the growth state. Both the specific activities of thymidine kinase and nucleoside phosphotransferase decrease steadily from the start of the experiments when the cell numbers were about 2-3 x 10(4) cells/ml in the PPL medium, while in the Neff's medium, the specific activities of thymidine kinase increase up to when the cell numbers reached 3-5 x 10(5) cells/ml and then decreased, but the specific activities of nucleoside phosphotransferase continuously decreased when the cell concentrations were 2-6 x 10(4) cells/ml. 4. In the PPL medium, the final cell numbers reached are about 6.5 x 10(5) cells/ml, while in the Neff's medium, the cell numbers increase further (to about 2 x 10(6) cells/ml). 5. No striking difference in activities of thymidine kinase and nucleoside phosphotransferase was observed when the cells were transferred from the defined medium to the Neff's medium, contrary to that reported by others for the activity of thymidylate synthetase.


The biosynthesis of nicotinamide adenine dinucleotide during early stages of frog embryonic development of haploid and diploid embryos. 1. Concentration of NAD during embryonic development of haploid and diploid embryos of frog was followed. NAD content in haploid embryonic forms is twice that in diploid embryos. 2. The variation of the NMN adenylyltransferase activity in the oocytes and during the first states of embryonic development as surveyed in the nuclear soluble fraction and the nuclear insoluble fraction (chromatin). 3. The enzyme activity in the soluble fraction is low during embryonic development and shows higher values in haploid embryos. 4. In the nonfertilized mature oocytes, the NMN adenylyltransferase activity is sixfold higher in the insoluble chromatin fraction than in the soluble fraction. 5. The evolution of the NMN adenylyltransferase in the insoluble chromatin fraction also shows higher values in haploid embryos, as compared with diploid forms.


Acid glycohydrolase in Chinese hamster (Cricetulus griseus) with spontaneous diabetes--V. Subcellular distribution in the kidney. 1. Seven renal glycohydrolases were measured in four subcellular fractions prepared from highly inbred aglycosuric (AV-line) and glycosuric (XA-line) Chinese hamsters. 2. alpha-D-galactosidase and beta-D-galactosidase were highest in the nuclear (N) and supernatant (S) fractions; both fractions showed reduced activities in the XA animals. 3. alpha-D-mannosidase was chiefly a particulate enzyme and its decrease in XA animals was evident in N, lysosomal-mitochondrial (LM) and mitochondrial-microsomal (MM) fractions. 4. No significant difference in N-acetyl-beta-D-glucosaminidase was found in any of these subcellular fractions between AV and XA animals. 5. Although total alpha-L-fucosidase and beta-D-fucosidase levels were similar in AV and XA kidneys, a difference was observed in the S fraction. 6. beta-D-glucuronidase was virtually absent in N and LM fractions and the S fraction of AV kidneys showed higher activity than the XAs.


Combination chemotherapy including high dose methotrexate and radiotherapy, in the treatment of small cell carcinoma of the lung. Twenty-nine (88%) of thirty-three patients who were treated with multiple drug chemotherapy, including high dose methotrexate, and radiotherapy for small cell carcinoma of the lung showed significant improvement in their clinical condition and quality of life. Treatment was well tolerated and toxicity acceptable. Cerebral metastases were not detected in any patient on presentation and only developed in three patients (9%). Little information exists regarding the use of high dose methotrexate in small cell carcinoma of the lung. There is no evidence, on the data available, that high dose methotrexate is any more effective than conventional doses.


Biological effects of chrysotile after SO2 sorption. II. Effects on alveolar macrophages and red blood cells. An experimental study has been carried out using different biological and biochemical in vivo and in vitro tests for assessing the toxicity of natural UICC (A) chrysotile and SO2-sorbed UICC (A) chrysotile and for detecting a possible synergistic effect. For in vivo studies, rabbits received an intratracheal injection of chrysotile fibers suspended in physiological saline (PS). The control group received only PS; all animals were sacrificed after 68 hr. The alveolar free cells were harvested by pulmonary lavage. The results have shown that chrysotile induces a decrease in the free cell population but there was no significant difference in the number of viable cells or in the nature of cells harvested between the two chrysotile groups. Enzymatic activities of the alveolar macrophages from animals injected with SO2-sorbed chrysotile showed a significant increase of the enzymes LDH and acid phosphatases. The LDH increase could be related to the affinity of the enzyme regarding some chemical forms of SO2. In vitro studies using alveolar macrophages harvested by pulmonary lavage have shown no differences between the two chrysotile groups when cell viability and enzyme release were studied. The toxic effect was due to chrysotile fibers (decrease of 38% in cell viability and increase in cellular enzyme release when compared with control). When rabbit red blood cells were used, both natural and SO2 chrysotile fibers showed the same hemolytic activity. The failure to detect high differences between the two chrysotile groups may be related to the chemical form of sorbed SO2.


Effects of asbestos on epithelioid cell lines. The in vitro effects of asbestos fibers on the growth and viability of CHO cells, an epithelioid cell line derived from Chinese hamster ovary, and on K-22 cells, an epithelial cell line derived from rat liver, have been studied. Relatively low concentrations of asbestos (10 micrograms/ml) were quite cytotoxic to both cell types. A sample of chrysotile asbestos was more toxic than samples of crocidolite or amosite. The toxic factor could not be extracted from the asbestos and toxicity occurred only if there was physical contact between the fibers and the cells. Although the phorbol ester class of tumor promoters induces the synthesis of plasminogen activator in various cell cultures, asbestos did not induce this protease in epithelioid or fibroblast cell cultures. The results obtained are compared to previous in vitro results obtained with fibroblast or macrophage cultures and are discussed in terms of their possible relevance to asbestos-induced fibrosis and tumorigenesis.


Low level lead and inhibition of erythrocyte pyrimidine nucleotidase. Pyrimidine 5'-nucleotidase (P5N, EC 3.1.3.5) appears to be a sensitive index of exposure to low level lead. In 21 children 2 to 5 years old with blood leads of 7 to 80 micrograms/dl there was a negative linear correlation of blood lead and red cell P5N: r = -0.60 (P less than 0.01), Y = -0.11X + 12.3. In rats, the enzyme assay was quantitatively similar to that of the human. A treatment group of 12 rats received lead acetate, 36 mg/kg/day, of lead as 0.17 M lead acetate for 24 days. The blood lead of treated rats increased from the control value of 8.3 +/- 1.3 to 36.0 +/- 0.5 on day 24; P5N decreased from 18.3 +/- 0.8 units to 9.0 +/- 1.0 and was below control values at a blood lead of 25. There was a significant negative linear correlation of blood lead and P5N: r = -0.85; n = 17; P less than 0.001; Y = 0.34X + 20.9 that was independent of the correlationship with the reticulocytes. At these levels of blood lead and P5N there was no significant change in the hexokinase, hemoglobin or red cell count and no evidence of stippling.


Mesothelioma after crocidolite exposure during gas mask manufacture. Of 199 persons employed in the manufacture and handling of Canadian military gas mask canisters containing pure crocidolite, 1939 to 1942, by the end of 1975, 56 had died, 120 were still alive, and 23 could not be traced. Nine (16%) of the deaths were probably due to malignant mesothelioma, six involving the peritoneum. The risk of mesothelioma after crocidolite exposure appears to be many times greater than that after chrysotile.


Kinetics of the potential-sensitive extrinsic probe oxonol VI in beef heart submitochondrial particles. The interaction of the potential-sensitive extrinsic probe oxonol VI with beef heart submitochondrial particles has been investigated under time resolved and equilibrium conditions. The time course of the probe absorption spectrum red shift induced by ATP or NADH injection into a suspension of submitochondrial particles in a dye solution is biphasic, consisting of a faster process described by a second-order rate law with k2 approximately 3 x 10(5) M-1 sec-1. For the ATP pulse experiments, the slower process follows first-order kinetics with k1 approximately 0.3 sec-1. In oxygen pulse experiments to an anaerobic dye-particle system, the slower process is not significantly developed due to rapid depletion of the oxygen, but the faster process follows second-order kinetics with the same rate of the oxygen, but the faster process follows second-order kinetics with the same rate constant as for the ATP and NADH cases. Evidence for permeation of the submitochondrial particle membrane by oxonol VI has been obtained; the slower process is interpretable as describing the permeation of the membrane bilayer. The results of the time-resolved work are consistent with a mechanism involving a redistribution of the dye from the bulk phase to the particle membrane. The value of the second-order rate constant for passive binding of the dye to submitochondrial particles is not compatible with a mechanism proposed to explain the microsecond probe response times in bilayer and excitable membrane experiments nor are such rapid signals observed in the oxonol VI-submitochondrial particle system.


Ontogeny and effect of vitamin D deprivation on rat serum 25-hydroxyvitamin D binding protein. Specific serum binding of 25-hydroxy-cholecalciferol (25-OHD3) was measured by saturation analysis in rats of various ages, and during vitamin D deprivation. The serum binding capacity for 25-OHD3 was observed to increase until age 6-8 weeks, then decline and remain stable thereafter at 3.7 x 10(-6) M. The 25-hydroxyvitamin D (25-OHD) concentration decreased in serum from rats fed vitamin D-free diet (t1/2 = 7 days). Rats fed 2 IU of vitamin D3/g of diet maintained stable serum levels of 25-OHD at 10-12 ng/ml. Serum binding capacity and affinity for 25-OHD3 was not affected by vitamin D deprivation or hypocalcemia. In addition, the binding affinity did not differ as a function of age (Kd = 3.3 x 10(-9) M). Since normal serum concentrations of 25-OHD in the rat are 2-5 x 10(-8) M, only 1-2% of the serum binding sites for this sterol are occupied under physiological conditions.


Evidence for a role of alpha-adrenergic mechanisms in regulation of episodic growth hormone secretion in the rat. The role of catecholamines in regulation of episodic GH secretion was investigated in the male rat. Administration of alpha-methyl-p-tyrosine (alpha-MT; 250 mg/kg ip) caused significant suppression of GH bursts and resulted in marked elevation of plasma PRL. Intravenous administration of apomorphine (.03 and .1 mg/kg) had no effect on decreased GH levels, whereas clonidine (150 micrograms/kg) restored GH secretion. Apomorphine significantly reduced PRL levels in alpha-MT-treated rats whereas, clonidine resulted in a further increase in PRL.


Cultured thyroid cell adenosine 3',5'-cyclic monophosphate response to thyrotropin: loss and restoration of sensitivity to iodide inhibition. Unlike in all other thyroid preparations, exposure of dog thyroid cells in long-term monolayer culture to iodide (10(-7) to 10(-3) M for up to 19 h did not blunt the subsequent adenosine 3', 5'-cyclic monophosphate (cAMP) response to thyrotropin (TSH) stimulation. This lack of effect of iodide was observed even when confluent thyroid cells were "follicularized" by the action of TSH in the culture medium. Preincubation of these cells in thyroxine (T4) and triiodothyronine (T3) was similarly without effect on the subsequent cAMP response to TSH. Study of thyroid cells during the early phase of primary culture demonstrated that inhibition by iodide (10(-4) M) of the cAMP response to TSH occurred after 7 h but was lost after 48 h of cell culture. This inhibitory effect of iodide was prevented by the inclusion of methimazole in the preincubation medium. As with iodide-insensitive cells, T4 and T3 were without effect on the cAMP response to TSH in iodide-sensitive thyroid cells. Exposure of iodide-insensitive thyroid cells to iodide-containing medium obtained after 2 h of incubation with dog thyroid slices, as well as to medium enriched with the 100,000 g supernatant fraction of homogenates prepared from these thyroid slices, did not restore the inhibitory action of iodide. However, iodide-sensitivity of the cAMP response to TSH was restored by preincubation of iodide-insensitive cells in 10(-4) M iodide plus an H2O2-generating system (glucose-glucose oxidase). These data suggest that T4 and T3 are not organic iodine inhibitors of the thyroid cAMP response to TSH. In addition, they provide evidence against the existence of a soluble, freely diffusible, organic iodine inhibitor of thyroid adenylate cyclase. The loss of sensitivity to iodide inhibition of adenylate cyclase that occurs in thyroid cells shortly after initiation of primary culture appears to be related to a defect in the cellular organification mechanism, possibly the H2O2-generating system.


Influence of dietary vitamin D3 on the circulating concentration of its active metabolites in the chick and rat. Plasma concentrations of 25-hydroxyvitamin D3 (25-OHD3) and 1 alpha,25-dihydroxyvitamin D3 (1 alpha, 25-(OH)2D3) in growing chicks and weanling rats were measured by a new radioreceptor assay to determine the effects of varying dietary levels of vitamin D3. The plasma concentration of 25-OHD3 fell from 14.1 ng/ml in 1-day-old chicks to undetectable levels after 3 weeks on a rachitogenic diet. Circulating 1 alpha,25-(OH)2D3 hormone also decreased from 8.9 ng/100 ml to undetectable levels at 3 weeks in these chicks. Chicks receiving an optimal supplement of vitamin D3 (1.4 IU/g diet) for three to four weeks had plasma 25-OHD3 and 1 alpha,25-(OH)2D3 levels of 21-35 ng/ml and 5.1-7.5 ng/100 ml, respectively. Nutritional supplementation with a 50-fold excess of vitamin D3 (70 IU/g diet) elicited a substantial increase in plasma 25-OHD3 to 87-130 ng/ml, while plasma 1 alpha,25-(OH)2D3 was not increased. Increasing dietary calcium from 1.4 to 2.8% did not alter the circulating level of vitamin D3 metabolites in chicks fed 1.4 IU of vitamin D3/g diet. Direct measurement of the renal 25-OHD3-1 alpha-hydroxylase in vitro, showed that lowering dietary calcium or exclusion of vitamin D3 stimulated the biosynthesis of 1 alpha,25-(OH)2D3, but raising calcium did not alter the enzyme activity. It is concluded that the circulating concentration of the 1 alpha,25-(OH)2D3 hormone in the chick is unaffected by abnormally high intakes of vitamin D3 or calcium, but the renal production of the hormone increases during vitamin D3 or calcium deprivation. Additional studies in rats fed a diet supplemented with either 2 or 1000 IU of vitamin D3/g verify that the circulating concentration of 25-OHD3 is markedly increased when the dietary intake of vitamin D3 is elevated. Moreover, 1 alpha,25(OH)2D3 is not increased under these conditions, but actually falls significantly when the dietary level of vitamin D3 is raised from 2 to 1000 IU/g. These studies in both the chick and rat indicate that dietary vitamin D3 excess enhances circulating 25-OHD3, probably because the vitamin D3-25-hydroxylase enzyme is not strigently controlled. The fact that the circulating 1 alpha,25-(OH)2D3 is not concomitantly increased may reflect either decreased synthesis or increased utilization of the 1 alpha,25-(OH)2D3 sterol.


Effect of wheat fiber on blood lipids, fecal steroid excretion and serum iron. Adding 36 g of wheat fiber for 3 weeks to the metabolically controlled diets of six subjects produced a significant increase in daily fecal weight from 70.8 g +/- 6.2 SEM to 217 g +/- 12.1; serum iron also fell by 21 micrograms/100 ml +/- 2.1 SEM (P less than 0.001) during the added fiber period (measured in five subjects) as did mean corpuscular volume and mean corpuscular hemoglobin. Fecal neutral steroid concentration (measured in four subjects) fell from 31 to 17.3 mg/g dry weight (P less than 0.05) but the change in neutral steroid output and in acid steroid concentration and output was not significant. No change was seen in the serum levels of cholesterol and triglyceride.


Surgical management of carotid body tumor. Carotid body tumor or chemodectoma is a slow-growing lesion; it should be diagnosed early to facilitate the technical surgical resection. When these lesions are large, the routine use of a carotid internal bypass shunt is advocated for their safe removal. With the newer vascular approaches, specifically the carotid internal bypass shunt, resection can be performed with acceptable risks. A patient with large bilateral carotid body tumors had successful resection of both these lesions.


The effects of miconazole on the ultrastructure of Candida albicans. Ultrastructural changes in Candida albicans induced by increasing concentrations of miconazole in vitro are described. Fungistatic concentrations (10(-8) to 10(-7) M) induced minimal morphologic changes at the cell periphery. At 10(-6) M cell volume increased and peroxisomes became numerous in the cell interior. The minimal fungicidal dose of 10(-5) M caused severe damage to most of the cell population and a total fungicidal dose of 10(-4) M caused total internal cellular necrosis even when cell walls remained intact. It is suggested that miconazole inhibits the peroxidative enzymes cytochrome c-peroxidase and catalase. Cell necrosis then results from peroxide accumulation.


Mechanism of enamel dissolution and its prevention. In this paper the mechanism of dental enamel dissolution is discussed. Special attention is given to the anisotropic solubility behaviour (hollow tube formation) of demineralized enamel crystallites. The combined evidence strongly indicates that this peculiar dissolution behaviour is caused by dislocations. The dislocations--linear lattice defects--are present in the crystallite center and are the cause of two active dissolution sites in each crystallite. Initial as well as later stages in the caries process are discussed. Experimental evidence is presented which indicates that the active sites can be inactivated by agents like EHDP and MFP. After inactivation, acid penetration is either prevented or strongly retarded.


Alterations in murine delayed type hypersensitivity responses by delta-8-THC and cannabinol. The ability of cannabinol and delta-8-tetrahydrocannabinol (delta-8-THC), two cannabinoid marihuana components, to modify delayed-type hypersensitivity (DTH) to sheep red blood cells (SRBC) was investigated in mice. Reduction of DTH reactivity by cannabinol required multiple, daily, postimmunization drug administration. Neither multiple preimmunization dosings nor a single postimmunization dosing was effective. Delta-8-THC produced moderate (13-33%) suppression of DTH reactivity only upon multiple, preimmunization treatments, whereas postimmunization administration only delayed peak DTH reactivity 24 hours without altering the absolute response level. These results indicate that delta-8-THC and cannabinol possess slight to moderate activity in the suppression of DTH.


Restoration by levamisole of rosette-forming cells inhibited by histamine, db-CAMP and adenosine. Db-cAMP and adenosine in vitro markedly inhibited the E-rosette formation of peripheral T-cells of healthy subjects. Histamine in vitro also inhibited the E-rosette formation, but only in patients with allergic disorders. Levamisole in vitro significantly restored the E-rosette formation of T-cells inhibited by db-cAMP, adenosine and histamine.


[Physiology of smell and taste].

[Physiology of smell and taste]. The functional organization of olfaction and taste are briefly discussed in morphological, physiological, biochemical and behavioural terms. Olfaction in animals serves often for long range navigational purposes whereas taste acts as a close up "last moment food checking system". Special attention is given to the coding problems in both systems. In taste, the spatial coding mode is prevalent. In olfaction a very complex coding system exists, which used temporal as well as spatial means and in which a whole set of olfactory nerve fibers is activated during the transmission of any specific odor information, each fiber of the set discharging at a specific impulse pattern. The role of the olfactory bulb is seen as an integrating center with the capabilities for short and long term information storage. The impact of von Békésy's microstimulation experiments on the physiology of taste is discussed. Research on taste modifiers such as gymnemic acid or of the taste modifying protein "miraculin" enrich our present understanding of the interaction between taste stimulants and the chemoreceptor sites in the taste buds.


Cytomegalovirus infection: a seroepidemiologic comparison of nuns and women from a venereal disease clinic. A seroepidemiologic study of prevalence of antibody to cytomegalovirus (CMV) was simultaneously done in four populations: group I, nuns working as nurses or school teachers; group II, women admitted to a upper socioeconomic private hospital; group III, women admitted to a lower socioeconomic county hospital, and group and IV women attending a veneral disease clinic. Groups II, III and IV, were not statistically different and showed an abrupt rise in antibody prevalence during your adulthood. Group I, however, did not show the expected abrupt rise in antibody prevalence during young adulthood, and the prevalence in this group was singificantly lower than that in the other three groups at all but the oldest age range. These differences could not be accounted for by race, socioeconomic status or respiratory exposure to CMV. The data suggest that there may be more than one mechanism of CMV transmission and that venereal or intimate salivary contact may be a significant mode of spread in adults.


Seroepidemiology of human papovaviruses. Discovery of virgin populations and some unusual patterns of antibody prevalence among remote peoples of the world. A total of 1544 sera from 28 diverse and mainly isolated populations were examined for HI antibody to BK virus. A few extremely isolated populations were found with negligible or absent exposure to the virus, but in most populations, remote or cosmopolitan, antibody appeared in increasing prevalence during early childhood and remained stable throughout adult life. Antibody acquisition and prevalence rates in individual families reflected that of the general population. Examined for HI antibody to JC virus were 393 sera from 9 of the 28 populations. Age acquisition and prevalence rates of antibody were similar to those of BK virus, but experience with the 2 viruses was found to occur independently in several population groups, i.e., high exposure to BK with low exposure to JC, or vice-versa. Examined for neurtralizing antibody to SV40 were 151 sera with and without BK HI antibody in individuals from several primitive populations. SV40 antibody, mainly in low titer, occurred in 35% of the BK-positive group, but only 5% of the BK-negative group, suggesting that infection with BK or a closely related virus is responsible for antibody directed against SV40 in most humans unexposed to known vaccine or monkey sources of SV40 infection.


Common misconceptions.

Common misconceptions. Certain misconceptions about the law are common among physicians. The purpose of the legal system is to resolve disputes rather than dispense "justice." The rules of law are neither stable nor clear. Truth cannot be recovered for purposes of litigation; therefore, the evidence alone must determine the merit of a case. Many factors other than merit may also contribute substantially to the outcome of litigation.


The legal setting in prescribing drugs.

The legal setting in prescribing drugs. By training and experience physicians generally prescribe drugs with only the patient in mind. In today's environment it is essential that the physician stop and think of himself also. We are all aware of malpractice problems, which begin in the physician's office or at the hospital and, if severe, end in court. The ophthalmologist must now consider what a disgruntled patient can find in writing to use in court. Hospital records and physician's charts are now just the beginning point. The Physician's Desk Reference (PDR), the published literature, the package insert, the promotional material of pharmaceutical companies, the internal records of drug companies, the pharmacists' records, and the data now obtainable from government agencies all provide happy hunting grounds for a competent plaintiff's attorney. The ophthalmologist needs to be keenly aware of these potentially adverse resources and how they can be used against him. He also needs to know the alternatives available to him. Once these matters are brought into clear perspective, both the patient and the ophthalmologist will be better off.


The neuro-ophthalmology trap: failure to diagnose. A failure to diagnose was the basis for most of a series of 35 malpractice suits filed against ophthalmologists and other physicians. All of these cases presented problems in neuro-ophthalmologic diagnosis. Selected summaries illustrate typical diagnostic pitfalls. "Creative" consultation reports may be one effective prophylactic measure against such legal actions.


Special problems in ophthalmic subspecialties. A study of 276 medicolegal cases disclosed the number in various ophthalmic subspecialties. The medical reasons for filing these claims were usually based upon omission or commission of acts fundamental to good basic ophthalmology, not the omission of sophisticated procedures. Among the lesions were that physicians should plan for the care of emergencies, use few types of drugs, order roentgenograms if suspicions arise, use consultants, stay well within one's level of competence, and document everything.


Comparison of state legislation regarding professional liability. The so-called malpractice crisis of the mid-1970s alerted physicians to the need for legislative involvement in order to achieve needed malpractice tort reform. There has been a positive effort to modify the medical liability situation in each of our 50 states. A review of this legislative activity from 1975 through 1977 is presented. It is anticipated that a new malpractice crisis may develop during the early 1980s. Actuarial data now being collected plus the effects of such innovations as arbitration, pretrial screening panels, collateral source rules, and periodic funding of malpractice claims may provide an equitable approach to any future malpractice tort reform.


Some medicolegal tips.

Some medicolegal tips. The purposes for each type of medical record are critical legal determinants for its content. Deposition is the most important single step in the litigation process. The uses for a deposition greatly exceed the mere discovery of fact. The weight given the testimony of witnesses by a jury is affected by many subtle factors, including professional demeanor, attitude, manner of dress, ability to communicate and recollect, and prior inconsistent statements.


Suprathreshold static perimetry in glaucoma and other optic nerve disease. Comparisons between automated suprathreshold static perimetry and manual kinetic perimetry were performed for 226 eyes with glaucoma or ocular hypertension, and 147 eyes with other optic nerve disease. Both techniques produced similar high detection rates for glaucomatous visual field defects, whereas suprathreshold static perimetry performed significantly better than kinetic testing in optic nerve disease other than glaucoma. It is concluded that automated suprathreshold static perimetry is an excellent quantitative screening technique for detecting visual field defects.


Clinical experiences with the use of an automated perimeter (Octopus) in the diagnosis and management of patients with glaucoma and neurologic diseases. This is a study of 59 patients, 42 with glaucoma and 17 with neurological disease, to compare the Octopus automated perimeter with the conventional Goldmann and Tubinger perimeters for detection and assessment of visual field defects. We believe the Octopus automated perimeter is an excellent method for screening patients suspected of glaucoma and can be expected to identify over 90% of the patients actually having visual field loss due to glaucoma. In patients with neurological disease, the Goldmann perimeter was superior to either the Octopus or Tubinger perimeters in providing clinically useful information.


Uveitis: effect of vitrectomy combined with lensectomy. Combined lensectomy-vitrectomy was performed on 25 eyes in 20 patients with cataracta complicata associated with uveitis from various etiologies. Postoperative follow-up averaged 15 months, with visual improvement in 24 of the 25 eyes. Primary cause of postoperative decreased vision was cystoid macular edema. Operative complications included retinal detachment and choroidal ischemia. The technique employed and the effect of lensectomy-vitrectomy on decreasing the number and severity of recurrent attacks of uveitis are discussed.


The treatment of orbital rhabdomyosarcoma with irradiation and chemotherapy. Fifty-eight patients with orbital rhabdomyosarcoma were treated with irradiation alone (25) or irradiation and chemotherapy (33) with follow-ups of 6 months to 14 years (mean 5.2 yr). At present, 74% are alive and 26% have died. Local control of the tumor was accomplished in 91% of cases. When local sinuses were invaded the survival rate was 55%. Chemotherapy appears to be of greatest value when disease is limited to the orbit. Irradiation or irradiation and chemotherapy should now be the treatment of choice for orbital rhabdomyosarcoma.


Diagnosis and management of cancer metastatic to the uvea: a study of 70 cases. Although metastatic cancer to the uvea is reported to be the most common intraocular malignancy, most ophthalmologists have had little experience with its diagnosis and treatment. This report describes our experience with the diagnosis and management of 70 patients with metastatic cancer to the uvea. Many patients were evaluated with modern diagnostic modalities such as fluorescein angiography, ultrasonography, and the 32P test when indicated. Thirty-one percent of patients had no history of previous malignancy, and the ocular complaints represented the first symptoms of systemic cancer. The ocular malignancy often simulated better-known ophthalmic entities, and the referring diagnosis was correct in only 38% of cases. This series, therefore, reflects the clinical problem confronting the practicing ophthalmologist. About one-half of the patients were treated with external beam irradiation to the involved eye(s), which often resulted in dramatic resolution of the tumor and visual return. Other patients had either no treatment, chemotherapy, or enucleation in selected instances.


Glaucoma and expulsive hemorrhage mechanisms in the Sturge-Weber syndrome. An eye from an adult with Sturge-Weber syndrome and glaucoma is described histologically following enucleation for massive choroidal hemorrhage. The specimen is noted to contain hemangiomas of the episclera and choroid as well as a partial angle cleavage abnormality. Histologic evidence reviewed herein supports the concept that infantile glaucoma associated with this syndrome may be associated with marked filtration angle cleavage deformities. Some Sturge-Weber patients who develop unilateral glaucoma later as juveniles or adults appear to have milder expressions of angle cleavage abnormality or a forme fruste of this angle disorder. Any surgical procedure for this glaucoma preferably should be one which minimizes or eliminates operative hypotony, thus avoiding a possible expulsive hemorrhage.


Automated pattern analysis of corneal endothelium. A proper understanding of the endothelial cell morphology of the cornea is of great significance to the corneal surgeon. Specular microscopy allows direct visualization of endothelial cell morphology and a proper analysis of these data needs automated computerized type of systems. Automated pattern analysis seems to offer a good option in this direction. From our experience it appears that such systems can be used with great advantage in the analysis of endothelial cell morphology. The data obtained from such analysis help to develop some indices for various facets of endothelial cell morphology.


Cinnamic aldehyde allergy.

Cinnamic aldehyde allergy. Positive patch test reactions to 2% cinnamic aldehyde were obtained from 2.8% of 34 males and 9.1% of 55 females. The 9.1% female reactivity may be traceable to perfumed cosmetics. Simultaneous Jasmin allergy is documented as is the occurrence of alpha-amyl cinnamic aldehyde in synthetic Jasmin. The relevance of these reactions is discussed. In two of the patients a proper diagnosis was not made from the original contact history. Only after a screening tray demonstration of cinnamic aldehyde allergy could a relevant history be taken from these patients.


Skin reactions to propylene glycol.

Skin reactions to propylene glycol. Propylene glycol (PG), ethylene glycol (EG), and polyethylene glycol 400 (PEG 400) were tested, as is, in a total of 1,556 cases of eczema using the chamber test method. All the year round, the number of positive reactions to PG was 12.5% to EG 4.9%, and to PEG 400 0.3%. A total of 30% of the positive reactions to PG were allergic in appearance. Also the new fatty alcohol-PG bases of Metosyn ointment and Topilar ointment as well as Metosyn ointment (fluocinonide) itself provoked reactions in a great number of patients with positive reactions to PG, as is. The reactions to PG were considered to be truly allergic in four cases. In them, positive reactions were elicited by testing with high dilutions of PG and by applying the glycol in the patients' armpits as an open test. It is concluded that PG and topical preparations containing it in high concentrations should not be used with occlusion, and that allergic reactions must be watched.


The patch test--who should and should not use it and why. Since its introduction into medicine by Joseph Jadassohn in 1895, the patch test has proven its supreme value in tens of thousands of cases and in research. But emphasis must be placed on the grave errors which can occur when the test is used improperly. Errors are common in the direction of both false positives and false negatives. Common causes of errors are described in detail. The conclusion is drawn that this valuable test had best be used only by those who have been adequately trained and who can apply it regularly on a sufficiently large number of selected patients.


Relative irritancy of free fatty acids of different chain length. Free fatty acids of human skin surface lipids have previously been implicated in the pathogenesis of acne vulgaris because of their apparent irritant and comedogenic properties. Prior studies on the relative irritancy of free fatty acids revealed the saturated C8 to C14 fatty acids and a C18 dienoic unsaturated fatty acid (linoleic) to be most irritating. Saturated free fatty acids from C3 to C18, and unsaturated C18 free fatty acids were applied daily under occlusive patch tests to human skin until detectable erythema appeared. The most irritating fatty acids were C8 through C12. Of the unsaturated fatty acids tested, only linoleic acid produced irritation.


Cinnamic aldehyde in toothpaste. 1. Clinical aspects and patch tests. Although cinnamon is known to cause dermatitis in bakers and confectioners, it has only rarely been reported as causing trouble in food or cosmetics. A newly-formulated 'spicy' toothpaste containing cinnamon as a flavouring agent was the cause of oral symptoms in eight patients referred to clinics in Buckinghamshire and in Malmö and in a further eight patients discovered subsequently. Similar symptoms and patch test results were observed independently at both centres. Positive reactions were obtained with 1% cinnamic aldehyde in 15 out of 16 patients tested. Only one patient reacted to a standard sample of balsam of Peru (25% in petrolatum).


Sensitivity to cinnamic aldehyde in a toothpaste. 2. Further studies. The authors carried out further studies to assess the significance of the findings of patients sensitive to cinnamic aldehyde in a toothpaste. Selected groups of patients in whom sensitivity to this might have been overlooked were patch-tested. With the exception of three patients, two with cosmetic sensitivity and one with hand eczema, the results were negative. The frequency of use of the original toothpaste was assessed among patients attending the clinic and this was followed by a circular letter to general practitioners and dentists inviting referral of patients with oral symptoms. As a result, five further cases, four of whom gave positive patch test reactions to cinnamic aldehyde, were discovered. The authors emphasize the importance of attempting to obtain accurate information as quickly as possible when 'new' or hitherto unsuspected allergens are implicated.


Contact dermatitis in patients with leg ulcers. The incidence of contact allergy to 63 substances, including active principles and excipients used in a variety of topical preparations, has been studied by patch testing in 306 patients being treated conservatively for stasis dermatitis with or without ulceration. Positive reactions to one or more test compounds were recorded in 177 patients. The main offenders among drugs were neomycin, sulphonamide and promethazine; among the excipients, they were parabens, lanolins and benzoyl peroxide. Frequent reactions were observed to certain substances of the 'para' group, benzocaine, diaminodiphenylmethane, aniline and paraphenylendiamine, with the first three acting mainly as indicators of cross-sensitization. The extension and spread of allergic dermatitis can also be produced by drugs administered systematically. Recognition of the allergens responsible for allergic contact dermatitis arising at the lower leg site is a necessary condition if chronicity and spread of the dermatitis are to be prevented. For the same reason it is important to restrict topical applications to non-allergic substances.


The histopathology of fiberglass dermatitis in relation to von Hebra's concept of eczema. A group of 36 normal control persons was patch-tested with glass fibers. The tested filaments had a length of about 3 to 5 mm and a diameter of 9 to 13 microns. The glass fibers were obtained from the normal production process. They were coated with cured epoxy resin. A group of 29 normal control persons was tested with the same type of fibers from which the epoxy finish had been burned off. Another group of 36 persons was patch-tested with ultrasmall glass globes with a diameter of 10 to 15 microns. Coated and uncoated glass globules were tested. On every person 5 patch tests with the same material were done. They were removed after 1, 5, 24, 48 and 72 h. On coated fiber, 12 of 36 tested people reacted with either a + or a ++ reaction. On the uncoated fibers a reaction could be noted in 10 out of 29 persons. The patch tests with coated as well as uncoated glass globules were all negative. A total of 60 biopsies was performed in those cases where + or ++ reactions were noted. The histological examination of the biopsies showed no differences between the reactions on coated and uncoated fibers. The skin reactions to glass fiber showed, besides microtraumatic changes, several histological characteristics of an eczematous reaction, like spongiosis and perivascular lymphocytic infiltration. The importance of these findings is discussed. It is emphasized that histological alterations are only one of the six points which must be taken into account when the diagnosis of eczema is considered.


Occupational dermatitis in a 10-year material. This study included 1,752 patients considered to have occupational dermatoses. The most common diagnosis was contact dermatitis. The dermatitis was of an allergic type in three-quarters of men and in half of women. One-fifth of the women with irritant contact dermatitis had an atopic history. Contact dermatitis was localized on the hands in 94% of women and in 84% of men. The most common allergens in men were chromium, rubber and plastic, and in women nickel, rubber and chromium. Chromium allergy occurred in four-fifths of the men in the building, metal and tanning industries. In one-fifth of the women, nickel allergy developed in cleaning work. Rubber allergy developed in the rubber industry in one-fifth of the cases. Half of the women with contact dermatitis were engaged in either nursing or cleaning work. A follow-up 2-3 years after treatment of 555 patients with contact dermatitis was completed by means of questionnaires. The eczema was healed in one-quarter of the patients, one-half had periodic symptoms, and one-quarter had permanent symptoms. The prognosis was the same for those who changed their work or stopped working as it was for those who continued their eczema-inducing work.


Nonspecific activation of murine lymphocytes. I. Proliferation and polyclonal activation induced by 2-mercaptoethanol and alpha-thioglycerol. The effect of 2-mercaptoethanol (2-ME) and alpha-thioglycerol (alpha TG) on proliferation and polyclonal activation of lymphocytes was studied in cultures of spleen cells from C3H mice. Inclusion in serum-free or serum-containing medium of the optimal concentration (5 x 10(-5) M) of either 2-ME or alpha TG resulted in highly significant uptake and incorporation of tritiated thymidine ([3H]TdR) into DNA and in morphological blast transformation. These phenomena were dose-dependent, with both lower and higher doses causing less marked effects. The kinetic peak of these responses was found to occur at day 3 of culture. Improved cellular viability could not explain these results, because by day 3 there was no significant difference in viability between cells cultured in the presence or absence of 2-ME. 2-ME evoked a proliferative response in cultures of congenitally athymic (nu/nu) spleen cells that exhibited a similar but lower dose-response profile compared with that of heterozygous (nu/+) littermates. Cultures of bone marrow-derived (B) lymphocytes, generated by treatment of spleen cells with rabbit antithymocyte serum and complement, incorporated [3H]TdR to a degree at least equal to that of normal spleen cell cultures. Thymus-dependent (T) cells did not support significant 2-ME, alpha TG, or Concanavalin A responses in the absence of serum. However, when cultured in 5% fetal calf serum, definite T-cell responses occurred, though always of a lower magnitude than B-cell responses in this system. When the enriched B-cell and T-cell preparations were co-cultured, a synergistic response was noted. Macrophage dependency of the 2-ME and alpha TG effect was shown to be minimal. It is likely that the greater effectiveness of alpha TG relative to 2-ME is due to differences in the chemical structure of these two thiol compounds. The advantages of utilizing 2-ME and alpha TG as probes in the study of lymphocyte activation are evaluated and their possible mechanisms of action are discussed.


Histocompatibility antigen-activated cytotoxic T lymphocytes. I. Estimates of the absolute frequency of killer cells generated in vitro. A sensitive procedure employing limiting dilution of activated lymphocytes and 51Cr release from highly labeled target cells was used to derive minimal estimates of the absolute frequency of cytotoxic T lymphocytes (CTL) present in a population of mouse lymphocytes activated to alloantigens of the major histocompatibility complex in bulk mixed lymphocyte cultures. From this figure (0.7-1.2%), the maximal rate of target cell killing could be calculated to be approximately 4 targets/CTL/hour.


Histocompatibility antigen-activated cytotoxic T lymphocytes. II. Estimates of the frequency and specificity of precursors. Using limiting dilutions of responding cells in mouse mixed leukocyte cultures, we obtained direct estimates of the minimum frequency of precursors of cytotoxic T lymphocytes (CTL.P) for a variety of antigens. Depending on the strain combination, there were as many as 4-15 CTL.P reactive to DBA/2 among 10(4) lymph node cells. Taking into account that only 5-10% of peripheral T lymphocytes have the potential to develop into cytotoxic T lymphocytes (CTLs) (6), this implies that at least 1-2% of all CTL.P are responsive to any given H-2 haplotype difference. Precursors of cytotoxic cells thus have the same high frequency of cells reactive to alloantigens of the major histocompatibility complex as found among proliferating cells in graft-vs.-host reactions and mixed lymphocyte interactions. The frequencies of CTL.P reactive to xenoantigens (rat) or trinitrophenyl-modified self were less than half the frequency of alloreactive CTL.P. A minority of the CTL.P specific for one H-2 haplotype were also reactive to a third party H-2 haplotype, presumably on the basis of recognition of shared determinants. By dilution of sensitized cells from single microcultures, it was shown that a single CTL.P undergoes a minimum of three to four cell divisions and generates at least 8-16 CTLs after antigenic activation.


Frequency of occurrence of idiotypes associated with anti-p-azophenylarsonate antibodies arising in mice immunologically suppressed with respect to a cross-reactive idiotype. Inoculation of rabbit anti-idiotypic (anti-id) antibodies suppresses the subsequent appearance of a cross-reactive idiotype (CRI) associated with the anti-p-azophenylarsonate (anti-Ar) antibodies of A/J mice. Such suppressed mice produce normal concentrations of anti-Ar antibodies which lack the CRI, but against which anti-id antisera can be prepared. The anti-Ar antibodies of an individual, suppressed mouse do not in general share idiotype with anti-Ar antibodies of other A/J mice, either suppressed or nonsuppressed. The present experiments were undertaken to quantitate several "private idiotypes" in a large number of hyperimmunized A/J mice. Anti-Ar antibodies of three mice, suppressed for the CRI, were labeled with 125I and subjected to isoelectric focusing. Four single peaks, that were over 90% reactive with autologous antiid, were randomly selected for use as ligands in a radioimmunoassay, and ascitic fluids containing anti-Ar antibodies from 181 A/J mice were tested as inhibitors. Two of the four idiotypes could not be detected in any mouse other than the donor. The concentration of the idiotype was less than 1 part in 1,250 to less than 1 part in 25,000 of the anti-Ar antibody population; these are minimum values. A third idiotype was detected in 3 of the 181 mice, but at very low concentrations. The fourth idiotype was present in 28% of the mice, again at a low concentration. The data support the existence of a very large repertoire of anti-Ar antibodies in the A/J strain and are consistent with a process of random somatic mutation for generating diversity in hypervariable regions. It is proposed that the cross-reactive idiotype may be controlled by a germ line gene or a gene related to a germ line gene through a small number of somatic mutations; and that the idiotypes that were not detectable in other mice were the products of genes that had undergone extensive mutations, with a low probability of recurrence in other mice.


Generation of both cross-reactive and virus-specific T-cell populations after immunization with serologically distinct influenza A viruses. Specificity of cytotoxic T-cell function was investigated for a range of different influenza viruses. T cells from mice immunized with A or B strain influenza viruses, or with vaccinia virus, showed reciprocal exclusion of cytotoxicity. Extensive cross-reactivity was, however, found for lymphocyte populations from mice infected with a variety of serologically distinct influenza A viruses, though serum antibodies did not cross-react when tested in a radioimmunoassay using comparable target cells as immunoadsorbents. This apparent lack of T-cell specificity was recognized for immune spleen cells generated after intraperitoneal inoculation of high titers of virus, and for mediastinal lymph node populations from mice with pneumonia due to infection with much less virus. The phenomenon could not be explained on the basis of exposure to the chicken host component, which is common to A and B strain viruses. However, not all of the virus-immune T-cell clones are cross-reactive. Competitive-inhibition experiments indicate that a considerable proportion of the lymphocyte response is restricted to the immunizing virus. Even so, the less specific component is significant. Also, exposure to one type A virus was found to prime for an enhanced cell-mediated immunity response after challenge with a second, serologically different A strain virus.


Genetic control of the immune response to nuclease. V. Genetic linkage and strain distribution of anti-nuclease idiotypes. Rat antisera raised against anti-nuclease antibodies from mouse strains A/J and SJL detect strain-specific idiotypic determinants related to the antigen-combining site. These antisera have been used to investigate the genetic linkage and strain distribution of the anti-nuclease idiotypes. Despite the existence of an H-2-linked immune response gene controlling the humoral response to nuclease, expression of the A/J anti-nuclease idiotype has been shown to be independent of genes in the H-2 region: the A/J idiotype was present in immune sera from strains A/J (H-2a) and A.BY (H-2b) but absent in sera from strains B10 (H-2b) and B10.A (H-2a). An analysis of the segregation of the A/J idiotype in offspring of the backcross (A/J x B10.A) x B10.A demonstrated linkage to the Ig-1e heavy chain allotype markers. In a small sample of backcross animals a very high apparent recombination frequency was observed, but further backcross analyses and progeny testing of putative recombinant animals will be required to substantiate this observation. Analysis of the A/J and SJL anti-nuclease idiotype markers in the BALB/c, CB.20, and BAB.14 strains indicate that these idiotypic markers may permit mapping of distinct variable region genes.


Studies of the receptor for phage A25 in group A streptococci: the role of peptidoglycan in reversible adsorption. Irreversible adsorption of a virulent phage, phage A25, to heat-killed streptococci, groups A, G, and A variant, has been achieved. Adsorption reflected the observed host range for phage A25 in that heat-killed group B cells were not able to inactivate the phage. Broken cells, cell walls, and peptidoglycan prepared from a group A strain K56 failed to adsorb the phage irreversibly, but retained the potential to carry out reversible adsorption. Experimental data including electron microscopy have demonstrated the specificity of reversible adsorption and have identified the peptidoglycan as a necessary cellular component of the receptor. The sensitivity of whole cells and purified peptidoglycan to muralytic enzymes suggests that the cell wall and peptidoglycan must be intact for optimal adsorption. In general the results are explained by postulating that adsorption of A25 phage particles to group A cells occurs by a two-step process; the first step involves recognition and reversible binding of the phage tail to the cell wall peptidoglycan, the second step is an irreversible reaction catalyzed by a yet unidentified cellular component which is destroyed when cells are ruptured.


Spontaneous regression of Friend virus-induced erythroleukemia. I. The role of the helper murine leukemia virus component. The RFV strain of the Friend virus complex induces an erythroleukemia that spontaneously regresses. The tropism of regressing Friend virus complex (RFV), which is conferred by its helper MuLV component, MuLV-RF, is different from that of the conventional virus strain, CFV. RFV is NB-tropic and CFV is N-tropic. Passage of nonregressing CFV through Fv-1 incompatible Swiss/ICR mice changed the tropism of CFV from N to NB and resulted in a virus strain which induced erythroleukemia that regressed. Passage of NB-tropic CFV back through Fv-1 compatible mice maintained NB-tropism and regression. Altering the quantity or type of helper MuLV in RFV complex by addition of Ri-MuLV inhibited regression in proportion to the amount of added Ri-MuLV. These studies indicate a relationship between a change in virus tropism to NB by passage in certain hosts (e.g., Swiss/ICR mice) and the ability of Friend virus to induce erythroleukemia that spontaneously regresses. MuLV-RF isolated from the RFV complex induced lymphocytic leukemia in newborn mice which regressed and caused the regression of CFV-induced erythroleukemia. MuLV-RF is NB-tropic, contains no spleen focus-forming virus (SFFV) activity and helps SFFV form spleen foci in genetically restrictive mice. Pseudotype viruses were prepared, consisting of MuLV-RF, or other MuLV's, and SFFV derived from FV-B. The pseudotype viruses each acquired the tropism of the MuLV used in rescue. The pseudotype prepared with MuLV-RF or another NB-tropic MuLV-F, but not the virus obtained by rescue with N-tropic MuLV-F, induced erythroleukemia that spontaneously regressed. These studies demonstrate that the ability of RFV to induce erythroleukemia that spontaneously regresses is due to its helper MuLV component.


Differences in pyrogen production by mononuclear phagocytes and by fibroblasts or HeLa cells. Phagocytosis of bacteria stimulates "professional" phagocytes to produce and release endogenous pyrogen (EP), the protein that mediates fever. To determine whether "nonprofessional" phagocytes also have this capacity, mouse and human fibroblasts and HeLa cells were cultured after ingestion of latex or chicken erythrocytes (CE), and EP release into culture supernate measured by mouse assay. No detectable pyrogen was released by these cell types after phagocytosis, whereas both latex and CE stimulated EP production by cultured mouse macrophages. These studies support the hypothesis that only professional phagocytes of bone marrow origin synthesize EP and induce fever.


Independent populations of primed F1 guinea pig T lymphocytes respond to antigen-pulsed parental peritoneal exudate cells. Thymus-dependent (T) lymphocytes from (2 x 13)F1 hybrid guinea pigs immunized to ovalbumin (OVA) in complete Freund's adjuvant can be stimulated to proliferate in vitro by antigen-pulsed peritoneal exudate cells (PECs) derived from either strain 2 or strain 13 donors. In this communication, we show that the population of primed F1 T lymphocytes which can be activated by antigen-pulsed strain 2 PECs is largely independent of the population of cells that can be activated by antigen-pulsed strain 13 PECs. This was demonstrated by both positive and negative selection procedures. In the former, T lymphocytes from OVA-primed (2 x 13)F1 donors were enriched by initial culture with OVA-pulsed strain 2 or strain 13 PECs for 1 wk. Cells selected by culture with OVA-pulsed strain 2 PECs responded well to OVA-pulsed strain 2 PECs and poorly to OVA-pulsed strain 13 PECs. If positive selection had been carried out with OVA-pulsed strain 13 PECs, the selected F1 T cells responded well to OVA-pulsed 13 PECs and poorly to OVA-pulsed 2 PECs. Negative selection was achieved by short term culture with antigen-pulsed PECs and by eliminating proliferating cells by treatment with bromodeoxyuridine and light. This procedure demonstrated that the population of primed F1 T lymphocytes which are responsive to OVA or to purified protein derivative of tuberculin can be divided into subpopulations uniquely responsive to antigen on either strain 2 or strain 13 PECs. Evidence was presented to indicate that this selective responsiveness was not the result of the action of alloantigen-specific suppressor cells. The results are considered in terms of current concepts of the genetic and molecular regulation of the interaction of PECs and T lymphocytes.


Experimental conditions for obtaining suppressor and helper effects on the primary in vitro immune response by lymphocytes activated by polyclonal T-cell activators. The effect of the polyclonal T-cell activators (PTA) Con A and PHA on the specific immune response to sheep red blood cells (SRC) was studied. Addition of PTA either enhanced or suppressed the anti-SRC response, and two variables were found to affect the results: time of addition of the PTA and the strength of the response in control cultures not given PTA. If the response was high, even suboptimal PTA concentrations induced suppressive effects, but if the control response was low, due to deficient batches of sera or because of the absence of serum, the addition of PTA increased the response or restored it to normal levels. Suppression could be obtained if the PTA were added before or at the same time as the antigen and required high (optimal) PTA concentrations. If addition was delayed for 12-24 h the suppressive effects disappeared and previously suppressive concentrations of the PTA now caused an enhanced response. Analogous results were obtained if preactivated lymphocytes were added to the cultures instead of soluble PTA. Neither Con A, PHA, or lymphocytes preactivated by these PTA suppressed the polyclonal response induced by LPS or PPD. Irrespective of the time of addition and the culture conditions, enhancement of the anti-SRC response occurred at lower PTA concentrations than suppression. It was concluded that suppressor T cells, if they exist, do not act on B cells, but rather on helper cells needed for induction of thymus-dependent responses. The findings in this system are not compatible with the existence of a specific subset of suppressor T cells, but rather with the notion that suppression is caused by too much help.


Antiviral protection by virus-immune cytotoxic T cells: infected target cells are lysed before infectious virus progeny is assembled. Virus-immune cytotoxic T cells can inhibit effectively growth of vaccinia virus in acutely infected target cells in vitro by destroying infected target cells before infectious virus progeny is assembled. Together with the fact that virus-specific T cells are demonstrable after 3 days, very early during infection, and with strong circumstantial evidence from adoptive transfer models in vivo, these data suggest that in some virus infections T cells may in fact act cytolytically in vivo to prevent virus growth and spread and be an important early antiviral effector mechanism.


Generation of T-helper cells in vitro. I. Cellular and antigen requirements. A sequential mouse cell culture system is described for the induction and assay of T-helper cells. Unprimed, cortisone-resistant, nylon wool-purified thymocytes cultured with adherent peritoneal exudate cells can be primed in vitro with soluble carrier protein to generate carrier-reactive helper cells. These cultured cells enhance the anti-hapten plaque-forming response of hapten-primed spleen cell cultures to hapten carrier conjugates. The culture conditions, cellular manipulations, and antigen requirements for the optimal induction of helper cells with these purified cell populations is presented. The active helper cell generated in this culture system is a thymus-derived cell which requires macrophages for its induction and must be proliferate in vitro before the manifestation of helper-cell function. Helper cells generated in vitro stimulate both carrier-specific and nonspecific enhancement of splenic anti-hapten responses. The carrier-specific and nonspecific enhancement can be distinguished by the requirement for antigen in the helper cell and spleen cell cultures, the dose of helper cells added to the spleen cell cultures, and by the requirement for additional splenic adherent accessory cell interactions.


A requirement for antigen-specific helper T cells in the generation of cytotoxic T cells from thymocyte precursors. Thymocytes cultured with irradiated, allogeneic stimulator cells yield no cytotoxic effector cells after a period in culture. If, however, a population of irradiated spleen cells syngeneic to the responder cells are added to these cultures, cytotoxicity is generated. The helper activity present in the irradiated syngeneic spleen cells was found to be mediated by a cell bearing theta antigens. Furthermore, it was found to be antigen specific; helper cells which were tolerant of the stimulator cell antigens were unable to help the thymocyte responder cells, although these tolerant cells did contain helpers specific for a third party antigen. These experiments are consistent with a requirement for associative recognition of linked determinants in the induction of killer precursors which is thus strictly analogous to the induction of B-cell precursors via collaboration with helper T cells. In more extensive studies, it was found that histoincompatible helper cells (H-2b, H-2p, H-2q) were able to help a cytotoxic T cell (H-2k) response to a third party stimulator cell antigen (H-2d); that is, the helper T cells which interact with cytotoxic T-cell precursors are not strain specific. It seems likely that the histocompatible helper cells induce killer precursors in an antigen-specific cooperation event similar or identical to normal syngeneic cooperation.


Prevention of IgG2a production as a result of allotype-specific interaction between T and B cells. BALB/c T cells, which can prevent normal C57BL IgG2a allotype (G2) production of Ig-congenic partner mice (C.B mice), are shown capable of preventing the growth and G2 production of a C.B plasmacytoma (CBPC 101). Such cytotoxic or suppressor T cells are clearly allotype-specific (G2 Tcs cells). And since CBPC 101 B cells do not require specific helper T cells in order to grow, we infer that G2-bearing B cells (normal or neoplastic) must be the direct target of G2 Tcs cells. This mode of T cell prevention of allotype production contrasts that reported for suppressor T cells in (BALB/c x SJL)F1 mice.


DNA-binding property of Sm nuclear antigen. Antibodies in the sera of patients with systemic lupus erythematosus reacted with a nuclear acidic protein called Sm antigen, and these antibodies were used as reagents to identify Sm antigen in preparative fractionation procedures. DNA affinity chromatography showed that Sm antigen was associated with nuclear protein fractions which had DNA-binding capacity. Evidence was also presented that Sm antigen showed preferential binding for single-strand DNA over double-strand DNA. These studies demonstrate that spontaneously occurring anti-nuclear antibodies in disease states may be used to study the properties of cellular proteins which are present in trace amounts.


Eosinophils and resistance to Trichinella spiralis. Mice infected with Trichinella spiralis were depleted of eosinophils by repeated administration of rabbit anti-mouse eosinophil serum. There was no effect on the spontaneous expulsion of adult worms from the small intestines, but the numbers of larvae in the muscles were almost doubled. It is concluded that eosinophils contribute to resistance to the systemic phase of trichinosis.


Parallel synthesis of immunoglobulins and J chain in pokeweed mitogen-stimulated normal cells and in lymphoblastoid cell lines. The synthesis of intracellular J chains was found to be closely associated with that of intracellular immunoglobulin, regardless of its class, during the process of B-cell differentiation. This parallelism between the synthesis of J chain and immunoglobulin was particularly evident in their coincident appearance in serial observations of pokeweed mitogen (PWM)-stimulated lymphocytes. The intensity of J-chain staining by fluorescent reagents in the stimulated cells synthesizing IgG was similar to that found in cells synthesizing IgA or IgM. Evidence was obtained that the presence of J chain in the IgG-producing cells did not reflect antecedent synthesis of IgA or IgM. T cells stimulated by phytohemagglutinin and PWM failed to show J-chain synthesis. Observations on lymphoid cell lines showed a similar parallelism between intracellular Ig and J-chain synthesis; no relation to surface Ig was found.


The immune response of allophenic mice to the synthetic polymer L-glutamic acid, L-lysine, L-phenylalanine. II. Lack of gene complementation in two nonresponder strains. The genetic control of the immune response of inbred strains of mice to certain antigens has been demonstrated to be governed by a set of Ir genes linked to the major histocompatibility complex (H-2) of mice (1,2). Until recently, the control was thought to be governed by single, dominant genes, located within the I region of the H-2 complex. Merryman et al. (3) originally demonstrated that the immune response to the synthetic terpolymer L-glutamic acid, L-lysine, L-phenylaline (GLphi) is under dominant, H-2-linked Ir gene control (4-7). This was shown both by crossing two nonresponder parental strains to produce responder offspring in the F(1) generation, and by the analysis of appropriate recombinant stains of mice. The two complementing genes have been mapped in the IA and IC regions of the H-2 complex, and have been termed beta and alpha, respectively (5,6). Thus, any strain of mouse may contain neither, one, or both genes. Only mice containing both genes are capable of responding to GLphi. It has been shown using F(1) hybrid and recombinant strains of mice, that the alpha- and beta-genes can complement each other in either the cis (on the same chromosome) or in the trans (on different chromosomes) position (8). In this paper we report the results of studies aimed at answering the question of whether or not the alpha- and beta- genes can complement each other when they are present in different lymphoid cells. To this end we have constructed allophenic mice composed of two nonresponder strains (A and C57BL/6), which show gene complementation in the F(1) generation. Allophenic mice are chimeras containing two cell types coexisting in a "normal" environment. The mice were tested for the specific cellular composition of the two parental cell types and were found to possess a complete range in the relative proportion of the two cell types. This report demonstrates that regardless of the mixture of cell types present in the allophenic mice, none of them were responders to GLphi. Thus no complementation of the alpha- and beta-genes is seen when the two genes are present in different cells.


Transplacental influence of the thymus.

Transplacental influence of the thymus. Peripheral blood lymphocyte levels attained in adult rats that had been thymectomized at birth were markedly reduced if the two or three immediately preceding generations of rats from which they were bred had been thymectomized as neonates. The capacity of thymectomized animals to survive to adult life was also drastically reduced if they were bred from thymectomized stock. Pregnancy as a result of mating with syngeneic or allogeneic males produced an ephemeral increase in peripheral blood lymphocyte levels of third and fourth generation thymectomized females. These observations are most readily explicable on the basis of a thymus-derived humoral influence acting directly or indirectly to influence the circulating lymphocyte population.


Cell surface antigens of human malignant melanoma. III. Recognition of autoantibodies with unusual characteristics. The sera of three patients with malignant melanoma showing reactivity with surface antigens of cultured autologous melanoma cells were analyzed by mixed hemadsorption and immune adherence assays in conjunction with absorption tests. In contrast to the melanoma-specific antigens demonstrated previously, the surface antigens detected by these sera occurred on a broad range of nucleated cells, both normal and malignant, from human, monkey, mouse, and chicken sources. Each serum had a characteristic pattern of reactivity in absorption tests, indicating the detection of distinct antigenic systems. Two sera showed auto-, allo-, and xenoreactivity, as well as the capacity to distinguish different cell populations in the same individual. The other serum reacted with an antigen apparently universally present on nucleated cells from a variety of species, but absent on erythrocytes. As these patients had been treated with chemotherapy, this may have played a role in the emergence of these broadly reactive autoantibodies.


Joint recognition by cytotoxic T cells of inactivated Sendai virus and products of the major histocompatibility complex. Cytotoxic T cells specific for Sendai virus were generated by culturing murine spleen cells in vitro together with UV-inactivated Sendai virus. In vivo immunization of donor mice with UV-inactivated Sendai virus resulted in an in vitro secondary response of increased magnitude. Cytotoxic activity was demonstrated in a short-term 51Cr-release assay, using syngeneic tumor cells which had been coated with inactivated Sendai virus by incubation at 4 degrees C for 30 min. The lysis of Sendai virus-coated target cells was restricted by the H-2 haplotype of the target cells, suggesting that the H-2 genes of the target cell contributed to the specificity of the lysis. Kinetic experiments showed that susceptibility to lysis by cytotoxic T cells specific for Sendai virus appeared within 30 min after coating target cells with inactivated virus. Furthermore, there was no detectable synthesis of new proteins in cells treated with UV-inactivated Sendai virus. For these reasons, we suggest that neither viral replication nor the synthesis of new proteins are necessary for the production of the antigen recognized by cytotoxic cells specific for Sendai virus. We infer that the virus-specific component on the target cells is probably a preformed virion antigen adsorbed onto or integrated into the cell membrane. These results imply that, if the cytotoxic T cell recognizes a single antigenic determinant specified both by viral and H-2 genes, this determinant is formed by the physical association of H-2 and Sendai virus antigens rather than by their alteration during the processes of synthesis.