Effects of aprotinin on renal function and urinary prostaglandin excretion in conscious rats after acute salt loading.

1. Aprotinin, a potent kallikrein inhibitor, was given to conscious rats with and without expansion of the extracellular fluid volume with isotonic saline. 2. In non-expanded rats aprotinin had no effect on arterial pressure, glomerular filtration rate (GFR), hippuran clearance, urinary flow rate, absolute sodium and potassium excretion or free-water clearance. 3. In volume-expanded rats aprotinin significantly reduced GFR, hippuran clearance, urine volume (V) UNaV, UKV and Cwater/GFR without effect on systemic arterial pressure. 4. Urinary immunoreactive prostaglandin E2 excretion significantly increased during the expansion phase but returned to below the control range during stable extracellular fluid volume expansion. 5. Aprotinin significantly suppressed urinary immunoreactive prostaglandin E2 excretion in non-expanded rats and in volume-expand rats during the expansion phase, but not during stable expansion. 6. The results suggest that the kallikrein-kinin system may contribute to changes in renal function during extracellular volume expansion. This action may not necessarily be associated with changes in renal prostaglandin E2 activity.


Lecithin-cholesterol acyltransferase and the lipoprotein abnormalities of parenchymal liver disease.

1. Detailed studies have been made of the plasma lipoprotein abnormalities in parenchymal liver disease to test the hypothesis that the abnormalities would correlate with plasma lecithin-cholesterol acyltransferase (LCAT) activity. 2. When LCAT was high, very-low-density-lipoproteins (VLDL) were normal in composition and had a normal pre-beta electrophoretic mobility. When LCAT was low, VLDL concentrations were greatly reduced. 3. With high LCAT low-density lipoproteins (LDL) were normal. The LDL particles found with low LCAT activity were of normal size but of abnormal composition, being triglyceride rich and cholesteryl ester poor. Regardless of LCAT activity LDL were present in normal amounts. 4. High-density lipoproteins (HDL) were normal in composition and electron-microscopic appearance when LCAT activity was high. When LCAT activity was low HDL were abnormal in composition and 'stacked discs' were seen on electron microscopy. 5. These results suggest that low LCAT activity may be the cause of at least some of the lipoprotein changes of parenchymal liver disease.


Brain angiotensin II binding and central [Sar1,Ala8]angiotensin responses in normal rats and the New Zealand strain of genetically hypertensive rats.

1. Specific angiotensin II (ANGII) receptor binding was measured in regions of the brains of the New Zealand gentically hypertensive and normal rats. 2. ANGII receptor binding was consistently lower in the septum, midbrain, thalamus and posterior medulla of the genetically hypertensive rats than in normal rats. 3. Blood pressure responses to intraventricular injections of ANGII and an ANGII antagonist [Sar1,Ala8]angiotensin were studied in conscious and pentobarbitone-anaesthetized genetically hypertensive and normal rats. In conscious rats no significant difference between the two strains of rat was detected. 4. In pentobarbitone-anaesthetized rats intraventricular injection of 40 microgram of [Sar1,Ala8]angiotensin had a hypotensive effect which was three times greater in the genetically hypertensive rats than that observed in normal rats. The latency of this hypotensive effect was longer than the latency of the hypertensive effect of ANGII. 5. The drinking responses to intraventricular injections of ANGII were similar in genetically hypertensive and normal rats. 6. The physiological role of the ANGII system is discussed and it is concluded that an abnormality of this system in the brain may well be responsible for the hypertension found in the genetically hypertensive rat.


DNA-dependent ATPases from Escherichia coli K12.

Four DNA-dependent ATPases have been isolated from E. coli extracts. ATPases I and III, both sensitive to NEM, require denatured DNA but differ in their heat sensitivity, elution from DEAE-cellulose, and sedimentation coefficient. ATPases II and IV are both resistant to NEM. ATPase II requires partially denatured DNA, whereas ATPase IV can be stimulated by SS DNA. ATPase I is a DNA-unwinding enzyme; ATPase II may be involved in recombination.


DNA helicases.

In summary, we postulate that DNA unwinding and ATP dephosphorylation are coupled in different ways, depending on whether the fibrous ATPase or one of the globular ATPases provides the catalytic agent. Unanswered is the question of whether there is stoichiometry of ATP utilization during the unwinding of a duplex, and unsolved is the role of the individual enzyme in the cell.


Changes in adrenocorticotropic hormone (ACTH) and cortisol levels in drug addicts treated by a new and rapid detoxification procedure using acupuncture and naloxone.

Plasma ACTH, cortisol, and cyclic-AMP levels of eleven heroin addicts were dertermined before and after treatment with a fast detoxification procedure using acupuncture and electrical stimulation (AES) together with the administration of limited doses of naloxone. At the end of the treatment period, the average plasma ACTH, cortisol, and cyclic-AMP level rose 130,83, and 24 percent, respectively. In view of the appearance of mild withdrawal signs during this method of treatment, the observed increases in ACTH and cortisol levels probably reflect the inability of AES to suppress withdrawal symptoms induced by naloxone completely. The mechanism underlying this new method of treatment is discussed in relation to AES's ability to stimulate the secretion of endorphins.


Prevalence of neutralizing antibody to the calf rotavirus in New York cattle.

In March 1973, a modified live virus vaccine was released for sale in the United States for the protection of neonatal calves from infection with calf rotavirus. At that time no published evidence existed that this agent was present in New York or the New England states. Serum neutralizing antibodies for the calf rotavirus (reovirus-like agent of neonatal calf diarrhea) were detected in serum from 108 of 110 dairy cattle in New York State representing 78 different herds. To exclude the possibility that the demonstrated serologic response may have been stimulated by vaccine virus, 36 samples were included that had been collected prior to the date of vaccine release. Neutralizing antibodies were demonstrated in 108 of the 110 sera ranging from titers of 4 to greater than 1024, thereby offering indirect evidence of the ubiquitous nature of calf rotavirus in New York.


Short-term immobilization of healthy men: right ventricular function and metabolism during graded exercise.

Five healthy male volunteers were subjected to graded exercise tests on a bicycle ergometer (600 kpm/min for 8 minutes) with simultaneous catheterization of the right cardiac ventricle and radial artery, first before immobilization and them after five-day bed rest in the head-down posture (the foot bed end was elevated 4.5 degrees). After immobilization, during exercise the stroke index did not increase, the tachycardia was more marked, the indicators of right ventricular contractility increased [+ max dp/dt, mas (dp/dt) /P, --mas dp/dt], the arterial pressure decreased, and the lactic acid level in mixed venous blood rose. Possible causes of haemodynamic and metabolic alterations, occurring in healthy volunteers during graded exercise following short-term hypokinesis, are discussed.


Simultaneous infection with Treponema pallidum and herpes simplex virus.

Two cases are presented of simultaneous infection with Treponema pallidum and herpes simplex virus. The occasional concurrence of these diseases suggests that all cases of herpes genitalis, including those with negative darkfield examinations and initial nonreactive serologic tests for syphilis, should be followed in two to four weeks by a repeat serologic test for syphilis.


Aniridia-Wilms' tumor association: evidence for specific deletion of 11p13.

A 7-year-old boy with aniridia, Wilms' tumor, and mental retardation, previously reported as having an interstitial deletion of the short arm of chromosome 8 resulting from a t(8p+;11q-) translocation (Ladda et al., 1974), has been restudied using high-resolution trypsin-Giemsa banding of prometaphase chromsomes. The results revealed a complex rearrangement with four break points in 8p, 11p, and 11q, leading to a net loss of an interstitial segment of 11p (region p1407 yields p1304) but not of 8p. His red blood cells contained normal activities of glutathione reductase (gene on 8p) and lactate dehydrogeanse A (gene on 11p12), indicating a gene dosage consistent with the chromosomal findings. The revised interpretation of this case agrees with seven others reported as having aniridia and interstitial 11p deletions in establishing the distal half of band 11p13 as the site of gene(s) which lead to aniridia and predispose to Wilms' tumor if present in a hemizygous state. Possible relationships between heterozygous deletion of specific chromosomal bands 11p13 and 13q14 and the autosomal dominant disorders aniridia, Wilms' tumor, and retinoblastoma, respectively, are discussed.


Clinical significance of pulmonary function tests. Alterations in pulmonary function following respiratory viral infection.

Respiratory viral illness is a major cause of morbidity in both adults and children. This report focuses on both the acute and chronic effects on respiratory function of these ubiquitous infections. Infant airways are particularly vulnerable due to the relatively low conductance in immature peripheral airways. Bronchiolitis, caused predominantly by respiratory syncytial virus, is the most important of these viral illnesses and is emerging as a major risk factor for the subsequent development of obstructive airway diseases in adults, possibly by interference with normal alveolar proliferation. The basic pathogenic mechanism involved in adult respiratory viral infection is bronchial hyperreactivity, presumably secondary to epithelial damage and resultant sensitization of rapidly adapting airway receptors. In addition, there may be virus-related alterations in the autonomic and humoral regulation of airway tone. Viral infections may alter the effects of common air pollutants on respiratory function.


Bronchial angiofibromata in a suspected case of tuberous sclerosis.

Widespread hamartomatous proliferations are often seen with tuberous sclerosis, yet pulmonary involvement is rare. We describe a patient with a probable forme fruste of tuberous sclerosis in whom the diagnosis of bronchial angiofibromata was made bronchoscopically. This is the first description of this pulmonary manifestation.


Attempts to standardize the screening for antiviral drugs by in vitro tests.

Plaque reduction (PR) and persistent infection cell culture (PICC) tests were used as a standard procedure for the selection of effective and well-tolerated antiviral compounds. Antiviral activity and cytotoxicity as well as other characteristics of the test compound, i.e., the development of drug resistance or the potential for activating the multiplication of other viruses can be examined by this method. In the described system analogues of 2'-deoxyuridine were tested against herpes simplex virus.


9-(2-hydroxyethoxymethyl)guanine as an inhibitor of herpes simplex virus replication.

The drug 9-(2-hydroxyethoxymethyl)guanine effectively inhibits herpes simplex virus replication. It is selectively phosphorylated by the virus-induced thymidine kinase but not by normal cellular thymidine kinase.


Therapeutic activity of pretazettine, a narcissus alkaloid, on spontaneous AKR leukemia.

A narcissus alkaloid, pretazettine hydrochloride (PTZ) has been shown to be active against spontaneous AKR leukemia. The long-term treatment with PTZ begining at 5--7 months of age of a group of AKR mice containing 10--20% of advanced leukemic mice significantly prolonged the life span of the group. The therapeutic effectiveness of PTZ has been compared with several standard antileukemic drugs. PTZ decreased the AKR virus titer in the circulating blood of mice and its antiviral activity in AKR virus infected NIH/3T3 cells has been confirmed by XC assay.


[New aspects in the concept of adjuvant and after-care therapy for gastric carcinoma?].

Basis and current possibilities of adjuvant therapy in gastric cancer are discussed. The response rate with 5-FU und nitrosoureas for advanced gastric cancer is significantly superior to all other regimens. Based on Japanese experiences a higher survival rate is observed in the curatively resected patients treated with postoperative prophylactic chemotherapy. The drawing of reliable conclusions must await the results of further studies. Efforts for individual chemotherapy by measurement of cell production of recurrences and the necessity of aftercare in gastric cancer patients are explained.


Dioxygen and the vitamin K-dependent synthesis of prothrombin.

It has been shown that bovine erythrocyte superoxide dismutase inhibits the gamma-carboxylation of glutamyl residues in the precursor of prothrombin and in a related synthetic peptide by a vitamin K-dependent rat liver microsomal carboxylase. In the same conditions a simple copper(II) tyrosinyl complex also inhibits the carobxylation. The formation of the vitamin K epoxide by the same systems is inhibited by both superoxide dismutase and catalase. It is suggested that the formation of the epoxide is a process distinct from carboxylation, representing perhaps a protective mechanism against the superoxide ion, or species derived therefrom, generated by the reaction of reduced vitamin K with dioxygen. Furthermore, the possibility that the carboxylating species is formed by the reaction of the superoxide ion, or species derived therefrom, with carbon dioxide, is proposed.


Oxygen consumption by stimulated human neutrophils.

Oxygen consumption by stimulated human neutrophils has been studied. An initial lag after stimulation of about 20 s is followed by a linear phase of oxygen consumption which lasts about 60 s and then declines exponentially. The duration of the respiratory  burst is thus much shorter than has been generally recognized. The cessation of the linear phase of oxygen consumption occurs at a constant time after stimulation, is not due to depletion of the substrate of the oxidase enzyme (since linear oxygen consumption can be re-initiated), and is due to termination or saturation of particle uptake. The curtailment of oxygen consumption probably marks the end of the oxygen-dependent microbial role of the phagocytic vacuole, preparing it for the secondary and independent function of digestion. A novel cytochrome b becomes associated with the phagosomes and probably forms a component of a complex electron-transport chain. Superoxide may be an intermediate in this system, but it is unlikely to be released free from the cell or to form the final product of the system.


Defects in the oxidative killing of microorganisms by phagocytic leukocytes.

One of the most important mechanisms of phagocytic killing of ingested microorganisms by leukocytes is the generation of toxic oxygen products. During phagocytosis, neutrophils, as well as monocytes and macrophages, display a strongly increased cell respiration. Quantitatively the most important product of this reaction is hydrogen peroxide. Superoxide is also generated in large amounts, probably as an intermediate in the formation of hydrogen peroxide. Indications exist that singlet oxygen and hydroxyl radicals are also formed in this process. Some of these oxygen products have microbicidal properties by themselves. The effect of hydrogen peroxide is greatly enhanced by the enzyme myeloperoxidase. Several dysfunctions of this sytem are known. In chronic granulomatous disease the enzyme system that produces superoxide is not operative. Thus, no superoxide or hydrogen peroxide is generated, leading to a severely decreased bacterial killing capacity. The exact molecular defects in the X-linked and the autosomal form are as yet undefined. Two variants are also known: lipochrome histiocytosis, with different clinical and histological manifestations, and a 'triggering defect' where only strongly opsonized particles trigger the respiratory burst. Myeloperoxidase deficiency leads to slightly decreased killing capacity, especially for yeasts. In glucose-6-phosphate dehydrogenase deficiency no oxygen radicals or hydrogen peroxide are produced because no equivalents for oxygen reduction can be generated in the hexose-monophosphate shunt. Deficiencies in the glutathione redox system also result in impaired phagocyte function, probably because the cells have to be protected against their own toxic oxygen products.


The role of myeloperoxidase in the microbicidal activity of polymorphonuclear leukocytes.

Myeloperoxidase (MPO), H2O2 and a halide form a powerful antimicrobial system effective against bacteria, fungi, viruses and mammalian cells. After phagocytosis, MPO is released into the phagosome from adjacent granules where it interacts with H2O2 generated either by leukocytic or microbial metabolism and a halide such as chloride or iodide to form agents toxic to the ingested organisms. Evidence for H2O2 and MPO participation in the microbicidal activity of polymorphonuclear leukocytes (PMNs) has been obtained from patients with neutrophil dysfunction. In chronic granulomatous disease, PMNs have a microbicidal defect associated with the absence of the respiratory burst. The importance of H2O2 deficiency in the PMN dysfunction is emphasized by its reversal by H2O2. PMNs which lack MPO also have a major fungicidal and bactericidal defect. Bactericidal activity is particularly low during the early postphagocytic period, after which the organisms are killed. Although emphasizing the importance of MPO-mediated antimicrobial systems particularly during the early postphagocytic period, these findings also indicate the presence of MPO-independent systems which develop slowly but are ultimately effective. The MPO-independent antimicrobial systems may be oxygen-dependent or oxygen-independent. The acetaldehyde-xanthine oxidase system has been used as a model of the MPO-independent, oxygen-dependent antimicrobial systems of the PMN. A microbicidal effect by this system was observed which was inhibited by superoxide dismutase, catalase and scavengers of hydroxyl radicals (OH') and singlet oxygen (1O2). The microbicidal activity of acetaldehyde and xanthine oxidase is increased considerably by MPO and chloride. The formation of ethylene from methional or 2-oxo-4-methylthiobutyric acid by PMNs has been regarded as evidence for OH' formation. We have found ethylene formation to be largely dependent on MPO and evidence for the initiation of ethylene formation by 1O2 has been obtained. Both the xanthine oxidase system and the MPO-H2O2-halide system convert diphenylfuran into cis-dibenzoylethylene, an effect which is compatible with, although not proof of, the formation of 1O2 by these systems.


The lactoperoxidase-thiocyanate-hydrogen peroxide antibacterium system.

Lactoperoxidase present in various secretions oxidizes thiocyanate (SCN-) in the presence of H2O2 to an unstable oxidation product--hypothiocyanite (OSCN-), which is bactericidal for enteric pathogens including multiple antibiotic resistant strains of E. coli. The system damages the inner membrane causing leakage and cessation of uptake of nutrient, leading eventually to death of the organisms and lysis. The possible involvement of O2- and 1O2 is discussed.


Phagocyte-produced free radicals: roles in cytotoxicity and inflammation.

The production of superoxide free radical, O2-, by metabolically activated phagocytes results in damage to the phagocyte which is manifested by the premature death of the cell in vitro. The cytotoxic agent appears to be formed by the reaction of superoxide with hydrogen peroxide, and is thought to be hydroxyl radical or a secondary radical thereof. In vivo two animal models of induced inflammation also appear to be largely dependent on superoxide production by phagocytes for the development of tissue damage manifested as oedema. Intravenously administered superoxide dismutase shows anti-inflammatory activity in these models, but only when so derivatized that it can remain in the circulation for longer periods of time. Catalase, or a catalase derivative, on the other hand, shows no anti-inflammatory activity in vivo.


The chemistry of dioxygen and its reduction products.

The electronic properties of dioxygen are discussed with reference to its reactions in biological systems. In particular, the importance of its reduction to the superoxide ion is stressed. The physical and chemical properties of the superoxide ion are described and their relevance to its function in biochemical processes is emphasized. The relationship to other intermediates such as the hydroxyl radical is discussed.


Superoxide dismutases: defence against endogenous superoxide radical.

Attempts to measure the rate of O2- production, in whole cells or in intact subcellular organelles, are frustrated by the endogenous superoxide dismutase (SOD). Streptococcus faecalis contains a single manganese-SOD which was isolated and used as an antigen in the rabbit. A precipitating and inhibiting antibody was obtained and used to suppress the SOD in crude lysates of S. faecalis. It allowed the demonstration that 17% of the total oxygen uptake by such lysates, in the presence of NADH, was associated with O2- production. O2- attacks unsaturated lipids and breaches the integrity of membranes. When the membranes are free of lipid hydroperoxides, then both O2- and H2O2 are required and singlet oxygen appears to be the proximal attacking species. When the membrane contains some lipid hydroperoxide, then O2- is itself sufficient and seems to generate an alkoxy radical, by reacting with the lipid hydroperoxide. It appears likely that attack on membranes is one of the reasons for the cytotoxicity of O2-. In Escherichia coli the manganese-SOD is derepressed by O2-. This enzyme is not made in the absence of oxygen and in aerobic conditions any change which results in enhanced production of O2- calls forth an increased synthesis of this enzyme. Increased levels of SOD, however achieved, correlate with greater resistance towards oxygen toxicity. It is generally true that respiring cells contain more SOD than non-respiring cells. Among obligate anaerobes there is a correlation between SOD-content and tolerance towards oxygen. It is not known whether the SOD in obligate anaerobes is a retained primitive characteristic or one recently acquired by plasmid transfer. There is an exception to the rule that copper-zinc-SOD is found in eukaryotes but not in prokaryotes, and that is the symbiotic bacterium Photobacterium leiognathi. This symbiont may have obtained the Cu-ZnSOD gene from the host fish.


The role of viral transformation and cytogenetic changes in viral oncogenesis.

A wide variety of DNA viruses and a more restricted family of RNA viruses can transform normal cells in vitro. Transformation means either immortalization and/or the appearance of certain phenotypic changes. Although it has been often inferred that in vitro transformation can be essentially equated with malignant transformation, increasing evidence indicates that the latter, reflected by tumorigenicity in vivo, requires additional cytogenetic changes. The evidence will be reviewed for EB virus-associated human malignancy (Burkitt's lymphoma) and the role of the 14q + translocation marker in human B-cell neoplasia. These findings point to an initiating role of viral transformation, reflected by in vitro immortalization, followed by a cytogenetic evolution where chromosome 14-associated changes are essential for the liberation of B lymphocytes from super-imposed controls. The contrast of tissue-associated, specific chromosomal changes that bring about malignant transformation after the initiating impact of different agents will be illustrated experimentally for murine T-cell lymphoma. Here, X-ray, DMBA and different virus (RadLV, Gross virus)-induced T lymphomas show the same chromosomal change: trisomy 15. It may be questioned whether viral transformation can ever lead to neoplasia in the absence of subsequent cytogenetic changes.


[Therapeutic and diagnostic problems of generalized acute suppurative periodontitis marginalis profunda].

Generalized periodontal abscesses with considerable increase in the volume and sensitivity of the gingiva, tooth loosening, and malaise accompanied by fever and weight loss were observed in 22 of 35 patients studied. The symptoms developed after antibiotic treatment for another disease. Local therapeutic measures alone were successful. Success was obtained only after institution of directed antibiotic therapy.


[Reversible alterations of peripheral nerve function in thyroid dysfunction (author's transl)].

In 7 patients with hyperthyroidism and 9 patients with hypothyroidism both thve before treatment and after a period of normal thyroid function of at least one year duration. As far as conduction velocity and relative refractory period are concerned there was no difference between hyperthyroid patients and normal persons, whereas in cases of hypothyroidism before treatment the conduction velocity was shown to be significantly reduced; moreover a prolonged relative refractory period and a decline in amplitude of the nerve action potential were found. These alterations of peripheral nerve function, interpreted as evidence of neuropathy, proved to be reversible when the thyroid function returned to normal.


[Electroencephalographic assessment of dosage and sleep-profile in a hypnotic Triazolothienodiazepine (author's transl)].

We 941, a Triazolodiazepine, has satisfying hypnotic properties. The optimum dosage lies nearly at 0.3 mg. In this dosage the sleep-begin is shortened, the number of nightly wakeness is decreased and the total sleep-time is extended, while the sleep-profile remains unchanged. A normal cycling can be taken as a sign of qualitative good sleep. The REM-percentage is diminished insignificantly and increases again under the continuated therapy. The percentage of deep sleep increases by 0.3 mg. The subjective presentations confirm the melioration of sleep. In higher dosages troubles occur as decrease of deep-sleep stage and deteriorations of feeling. Remarkable is the withdrawal effect after 1.0 mg with an initial and a dissociated REM. However, such strong disturbances couldn't be observed in the carried on therapy with 0.3 mg.


Stimulation of guanosine 3',5'-monophosphate-phosphodiesterase activity by adrenocorticotropic hormone-activated increase of guanosine 3',5'-monophosphate in isolated adrenocortical carcinoma cells.

The decline of the ACTH-stimulated cGMP to the basal level in isolated adrenocortical carcinoma cells was inhibited by cyclic phosphodiesterase inhibitors. Time-course experiments showed that the ACTH-induced level of cGMP preceded the activation of phosphodiesterase. Cells incubated with increasing concentrations of exogenous cGMP responded with a corresponding increase in the cGMP-phosphodiesterase activity. cAMP was 100-fold less effective than cGMP in the activation of cGMP-phosphodiesterase, indicating the nucleotide specificity of enzyme activation. The activation of cGMP-phosphodiesterase by ACTH-induced cGMP or exogenous cGMP was rapid. In contrast to these observations, there was no activation of cAMP-phosphodiesterase by exogenous cAMP or cGMP. These results indicate that one mechanism by which isolated adrenocortical carcinoma cells regulate the ACTH-induced increase in cGMP is the cyclic nucleotide control of the activity of its phosphodiesterase, thereby regulating cGMP degradation.


Acute self-suppression of corticosteroidogenesis in isolated adrenocortical cells.

The relation between steroidogenesis induced by ACTH and that induced by exogenous concentrations of glucocorticoids was studied in isolated adrenocortical cells. Exogenous corticosterone and cortisol, in concentrations within the production capacity of the adrenal gland, suppressed steroidogenesis induced by ACTH in rat and beef cells, respectively. The precursors pregnenolone and progesterone enhanced steroidogenesis in both rat and beef cells. Aldosterone in rat cells and 17 beta-estradiol in rat and beef cells had little if any effect on steroidogenesis. Either suppression or stimulation by exogenous steroids was acute, that is, after 2-h incubation for rat cells and 1-h incubation for beef cells. A direct suppressive action of end product glucocorticoids is indicated. This observed self-suppression of adrenocortical cells suggests the existence of a mechanism for the find adjustment of steroidogenesis that operates in addition to the classical control exerted by the anterior pituitary.


Effect of calcium administration on renal responsiveness to parathyroid hormone in pseudohypoparathyroidism type I and II -- in comparison with normals, idiopathic and surgical hypoparathyroidism.

A 31-year-old man and a 12-year-old girl were diagnosed as pseudohypoparathyroidism (PHP) Type I because of a failure to respond to the administration of parathyroid hormone (PTH) with increased urinary excretion of phosphate and cyclic adenosine-3', 5'-monophosphate (cAMP). A 22-year-old woman was diagnosed as PHP Type II because there was no increase in the urinary excretion of phosphate despite of a marked increase in urinary cAMP excretion. With the combined calcium-PTH infusion or PTH infusion after vitamin D therapy, renal response was improved in these patients. Also dibutyryl adenosine-3'-5'-cyclic monophosphate (dbcAMP) infusion evoked an increased urinary phosphate excretion in all of the patients. The metabolic defect of our patients with PHP Type I may be caused not by a lack or defective form of PTH-sensitive receptor adenylate cyclase complex but rather by an abnormal conformation in the plasma membrane-associated receptor adenylate cyclase enzyme complex in kidney. In the patient with PHP Type II, as cAMP generation is intact, the metabolic defect might be related to a defect of calcium mobilization in renal tubular cells in response to PTH.


Effects of ACTH and calcium on cyclic AMP production and steroid output by the zona glomerulosa of the adrenal cortex.

Effects of ACTH and calcium on cyclic AMP production and steroid output by the zona glomerulosa (the capsular fraction) from the rat adrenal cortex have been studied. Although high concentrations of extracellular calcium potentiated the stimulatory action of ACTH on cyclic AMP and aldosterone output, tetracaine or verapamil inhibited aldosterone output but not cyclic AMP production during ACTH-stimulation. Lanthanum reduced both aldosterone and cyclic AMP accumulation induced by ACTH. These results suggest that an extracellular calcium would be essential in stimulating the capsular steroidogenesis without involvement of the cyclic AMP system.


Calcium ion as "second messenger" in corticoidogenic action of ACTH.

The correlation between corticoidogenesis and Ca2+-influx in the cell was investigated using isolated rat or bovine adrenocortical cells. 1) ACTH-induced corticoidogenesis in both rat and bovine adrenocortical cells was enhanced in parallel with increasing concentration of external Ca2+. In the bovine cells but not in the rat cells, moreover, the marked stimulatory effect of external Ca2+ on the corticoidogenesis was observed despite the absence of ACTH. 2) Ca2+-influx and corticoidogenesis always occurred unitedly. 3) Verapamil markedly inhibited either corticoidogenesis or Ca2+-influx in response to ACTH. 4) The stimulatory effect of Ca2+ on corticoidogenesis was completely blocked by cycloheximide. It was therefore suggested that Ca2+ could regulate corticoidogenesis as a primary "second messenger" of ACTH through biosynthesis of so-called steroidogenic protein.


Effects of vasopressin and cyclic AMP on water transport at arachnoid villi of cats.

The effects of vasopressin and cyclic AMP on water transport at arachnoid villi into the superior sagittal sinus were examined using the isolated meninges preparations of cats. The meninges preparation, the superior sagittal sinus of which was opened at the midline of the outer surface, was held between two polyethylene tubes. The tubes were fixed vertically in the way that the opened surface of the sinus was directed downward and arachnoid surface upward. Water transport was determined by measuring the tritiated water dripping through the membrane preparation. Vasopressin from less than 50 to 500 muU/ml accelerated the water transport and this effect was dose-dependent. Cyclic AMP from 0.5 to 10 mM was proved to manifest the same effect as vasopressin. This effect of cyclic AMP appeared rapidly in comparison with that of vasopressin, suggesting that the effect of vasopressin may be manifested through cyclic AMP. From these findings a physiological role of vasopressin in cerebrospinal fluid was discussed regarding the regulation of intracranial pressure.


Inactivation of human thyroid stimulator by human thyroid extract.

To determine whether the thyroid stimulating activity of IGG of patients with Graves' disease is associated with the reaction with a putative human thyroid antigen, the inactivation of the property of IgG to stimulate cAMP generation in human thyroid slices incubated in vitro was studied by pretreating the IgG with human thyroidal particulate fraction. In the preliminary experiment, it was demonstrated that to cause cAMP generation stimulation, on incubation period of 120 min is required to allow the IgG to penetrate the tissue. When human thyroid slices were incubated with normal IgG without or with pretreatment by human thyroid particulate fraction obtained from 100 mg tissue, cAMP content in the slices was 142 +/- 25 or 138 +/- 26 f moles/mg, respectively, indicating that basal thyroidal cAMP levels were not influenced at all by normal IgG even after pretreatment with thyroid particulate fraction. When the slices were incubated with gG of Graves' disease without or with the similar pretreatment, cAMP content was 320 +/- 31 or 140 +/- 25 f moles/mg, respectively, demonstrating an almost complete inhibition of the activity of the IgG to cause cAMP generation stimulation.


A case of histiocytosis X associated with panhypopituitarism and hyperimmunoglobulinemia G and E.

A 43 year old man with diabetes insipidus who showed panhypopituitarism and marked hypergammaglobulinemia due to histiocytosis X is reported. His low basal plasma adrenocorticotropin (ACTH) and growth hormone (GH) failed to respond to insulin-induced hypoglycemia. His basal serum thyroid hormone level was below normal and normal basal plasma thyrotropin (TSH) showed a delayed response with normal peak value to TSH-releasing hormone (TRH). Normal basal plasma pituitary gonadotropin also showed a delayed response with normal peak value to luteinizing hormone-releasing hormone (LH-RH). Suppression of plasma prolactin (PRL) by levodopa (l-dopa) was impaired and elevation of basal plasma PRL was noted at the second admission. These results, combined with diabetes insipidus, suggested that the panhypopituitarism in these patients was hypothalamic in origin. The polyclonal hypergammaglobulinemia was characterized by elevated serum IgG and IgE levels which returned to normal after corticosteroid treatment with concomitant clinical improvement. Elevated serum IgE levels, tissue and peripheral eosinophilia, and the effectiveness of corticosteroid therapy support the hypothesis that some allergic mechanism may be involved in the pathogenesis of this disease.


C-peptide, insulin and proinsulinlike components in diabetic and nondiabetic human pancreas.

The contents of insulin and C-peptide extractable with acid alcohol from the tail of the pancreas and insulinoma were investigated, using gel filtration in seven nondiabetics including two patients with insulinoma and eight diabetics. The gel filtration patterns of both C-peptide and insulin in pancreatic extract were fairly stable even after the pancreas had been left for 14 hrs in the room temperature. In nondiabetics except cases of insulinoma the content of insulin in pancreas ranged from 1.42 to 4.56 U per gram and that of C-peptide from 8.76 to 25.63 microgram per gram wet pancreas. The proportion of proinsulinlike components (PLC) ranged from 0.01 to 2.04% of insulin plus PLC. In diabetics insulin content was low and ranged from 0 to 1.68 U per gram and that of C-peptide from 0 to 14.48 microgram per gram wet pancreas. In insulinoma, both insulin and C-peptide increased and PLC occupied 5.48 and 5.96%, respectively.


Insulin release and metabolism of calcium, adenine and adenosine 3',5'-cyclic monophosphate.

In order to assess further the mechanisms involved in insulin release, we prelabeled rat pancreatic islets of Langerhans by incubating either 45Ca or [2-3H]adenine. When prelabeled islets were perfused with a glucose-free medium (the experiment with 45Ca) and a medium containing 2.8 mM glucose (the experiment with [2-3H]adenine) respectively, a constant rate of efflux of the radioactivity was established by 30 min in each case. D-Glucose at 16.7 mM concentration elicited a rapid efflux of 45Ca and [2-3H]adenine derivatives ([3H]Ad) within 4 to 6 min after commencing the step-wise stimulation by glucose, concomitantly with insulin release. However, L-glucose and D-galactose littel stimulated both 45Ca and [3H]Ad release. Lanthanum chloride caused a burst peak of 45Ca release in the absence of glucose. A rapid efflux of 45Ca was caused by beta-D-glucose and D-glyceraldehyde to much lesser extent than by alpha-D-glucose. The slowly rising concentration of glucose at 0.1 mM/min of gradient level failed to elicit any rapid efflux of 45Ca or [3H]Ad, although insulin release occurred in accordance with an increase in glucose concentration. Even when the gradient of glucose concentration was raised to 0.7 mM/min, glucose failed to stimulate an efflux of [3H]Ad but the subsequent stimulation by 16.7 mM glucose caused a rapid efflux of [3H]Ad concomitantly with the release of insulin. No rapid efflux of 45Ca was observed under a slow-rise glucose stimulation until the gradient level of the glucose concentration was raised to 6.7 mM. Analysis of distribution of the radioactive adenine derivatives after incubation showed that the adenosine fraction had the highest radioactivity in the medium followed by the ATP, adenine and cAMP fraction in that order, and the ATP fraction had the highest radioactivity in the islet. The ratio of radioactivity in the cAMP fraction in the medium to the total count was the highest among all. On the basis of these results, it was suggested that the discharge of [3H]Ad and 45Ca might occur with the alteration of the membrane permeability induced by a rapid change of the glucose concentration, and that their discharge might perhaps link to the glucoreceptor mechanism directly controlling insulin release.


The hormonal control of glycogen metabolism. Phosphorylation of protein phosphatase inhibitor-1 in vivo in response to adrenaline.

Inhibitor-1 has been shown to be phosphorylated in skeletal muscle in vivo. In normal fed animals the degree of phosphorylation was 31 +/- 7% and this value increases to 70 +/- 12% following an intravenous injection of adrenaline. The results imply that the phosphorylation of inhibitor-1 may be equally as important as the phosphorylation of phosphorylase kinase in elevating the levels of phosphorylase a. The role of inhibitor-1 in metabolism is discussed.


Nucleotide sequence of three isoaccepting lysine tRNAs from rabbit liver and SV40-transformed mouse fibroblasts.

The lysine isoacceptor tRNAs differ in two aspects from the majority of the other mammalian tRNA species: they do not contain ribosylthymine (T) in loop IV, and a 'new' lysine tRNA, which is practically absent in non-dividing tissue, appears at elevated levels in proliferating cells. We have therefore purified the three major isoaccepting lysine tRNAs from rabbit liver and the 'new' lysine tRNA isolated from SV40-transformed mouse fibroblasts, and determined their nucleotide sequences. Our basic findings are as follows. a) The three major lysine tRNAs (species 1, 2 and 3) from rabbit liver contain 2'-O-methylribosylthymine (Tm) in place of T. tRNA1Lys and tRNA2Lys differ only by a single base pair in the middle of the anticodon stem; the anticodon sequence C-U-U is followed by N-threonyl-adenosine (t6A). TRNA3Lys has the anticodon S-U-U and contains two highly modified thionucleosides, S (shown to be 2-thio-5-carboxymethyl-uridine methyl ester) and a further modified derivative of t6 A (2-methyl-thio-N6-threonyl-adenosine) on the 3' side of the anticodon. tRNA3Lys differs in 14 and 16 positions, respectively, from the other two isoacceptors. b) Protein synthesis in vitro, using synthetic polynucleotides of defined sequence, showed that tRNA2Lys with anticodon C-U-U recognized A-A-G only, whereas tRNA3Lys, which contains thio-nucleotides in and next to the anticodon, decodes both lysine codons A-A-G and A-A-A, but with a preference for A-A-A. In a globin-mRNA-translating cell-free system from ascites cells, both lysine tRNAs donated lysine into globin. The rate and extent of lysine incorporation, however, was higher with tRNA2Lys than with tRNA3Lys, in agreement with the fact that alpha-globin and beta-globin mRNAs contain more A-A-G than A-A-A- codons for lysine. c) A comparison of the nucleotide sequences of lysine tRNA species 1, 2 and 3 from rabbit liver, with that of the 'new' tRNA4Lys from transformed and rapidly dividing cells showed that this tRNA is not the product of a new gene or group of genes, but is an undermodified tRNA derived exclusively from tRNA2Lys. Of the two dihydrouridines present in tRNA2Lys, one is found as U in tRNA4Lys; the purine next to the anticodon is as yet unidentified but is known not be t6 A. In addition we have found U, T and psi besides Tm as the first nucleoside in loop IV.


Characterization of an adenosine triphosphatase from myeloblasts infected with the avian myeloblastosis virus.

An adenosine triphosphatase (ATPase EC 3.6.1.3) was partially purified from myeloblasts of chicken infected with the avian myeloblastosis virus and some of its molecular, catalytic and immunological properties were compared with that of the ATPase purified from the virus. Both the enzymes possessed almost same electrophoretic mobility, molecular weight, S20,w value, substrate specificity, metal-ion requirement, apparent Km value and sensitivity to inhibitors and activator. Evidence also indicated immunological identity of the two enzymes. The insensitivity of this enzyme to rutamycin or ouabain and extreme sensitivity to most of the detergents, trypsin and mercurials are the remarkable properties of this enzyme.


Cyclic 3',5'-adenosine monophosphate (cAMP) in a 42-month-old child with clinical evidence of multiple hormonal disturbances.

In a boy aged 42 months, small stature, retarded psychomotor development, dry skin, excessive thirst, polyuria, cryptorchidism, and rickets were signs of multihormonal disturbances. Contrary to the clinical manifestations, laboratory investigations showed normal or raised levels of hormones (hGH, insulin, T3RU, T4, TSH, PTH). The cAMP level in the plasma was low and its urinary excretion was reduced. After administration of hGH, adrenaline, T3, T4, pitressin, vitamin D3 and aminophylline there was no rise in the cAMP concentration in plasma and urine. In the light of these results it may be assumed that deficient function of the adenyl cyclase system led to development of a clinical syndrome of tissue insensitivity to multiple hormonal factors in this case.


Reduced high density lipoprotein in stroke: relationship with elevated triglyceride and hypertension.

Lipids and lipoproteins were analysed in forty-one survivors of stroke, aged less than 65 years, and the same number of age and sex matched controls without vascular disease. The stroke subjects had no evidence of coronary artery or peripheral vascular disease. High density lipoprotein cholesterol was significantly lower (1.19 +/- 0.06 mmol/l) in the stroke subjects than the controls (1.47 +/- 0.07 mmol/l). Triglyceride was also elevated in the stroke subjects, but this was confined to those who were taking antihypertensive treatment which included beta-blockers and/or thiazides. The low levels of high density lipoprotein in stroke were independent of hypertension or its treatment. Thus low levels of high density lipoprotein appear to be associated with cerebrovascular disease, while elevated triglyceride is a complication of anti-hypertensive therapy.


Selective labeling of apomorphine receptors by 3H-LSD.

There are at least two types of dopamine receptors: the 3H-dopamine or 3H-apomorphine receptor (with high or nM affinity for dopamine), and the 3H-neuroleptic receptor (with low or microM affinity for dopamine). While 3H-LSD can label the 3H-neuroleptic receptor, this study was done in order to label the 3H-apomorphine/dopamine receptor site. In the presence of excess phentolamine, serotonin and spiperone (to preclude binding to alpha-adrenergic, serotonergic and neuroleptic receptors, respectively) similar concentrations of dopaminergic drugs inhibited the binding (to calf caudate) of 3H-LSD and 3H-apomorphine. This is compatible with the concept that the 3H-apomorphine/dopamine receptor and the 3H-neuroleptic/dopamine receptor are separate.


Effect of chronic ethanol treatment on specific [3H]ouabain binding to (Na+ + K+)-ATPase in different areas of cat brain.

Chronic ethanol administration to cats increased specific [3H]ouabain binding by 63% in cerebral cortex, 47% in cerebellum, 84% in amygdala, and 100% in hippocampus when the binding assays were performed in the presence of 160 nM [3H]ouabain. There was no significant change in specific [3H]ouabain binding in hypothalamus, thalamus, corpus striatum, and brain stem following chronic ethanol ingestion. Scatchard analysis revealed that enhancement of specific [3H]ouabain binding following chronic ethanol treatment in some areas of cat brain is primarily due to changes in densities of ouabain binding sites. Since ouabain is a specific inhibitor of (Na+ + K+)-ATPase the present observations suggest that the molecular mechanism for the enhancement of (Na+ + K+)-ATPase activity after chronic ethanol ingestion may be due to increased net rate of synthesis of (Na+ + K+)-ATPase molecules or exposure of non-functional enzyme system following conformational change of plasma membrane.


GABA receptor-mediated modulation of 3H-diazepam binding in rat cortex.

Three membrane preparations of rat cortex were used to examine the effects of GABA, bicuculline and bicucine on specific 3H-diazepam binding. In the crude synaptosomal fraction, GABA had no effect on either the maximal binding capacity (Bmax) or dissociation constant (KD) of 3H-diazepam binding. Bicuculline and bicucine both decreased binding affinity. This was antagonized by adding GABA. In the repeatedly washed membrane preparation, and in the washed, frozen and thawed preparation, GABA increased binding affinity and, at high concentrations, increased Bmax. Increased binding affinity was observed with as little as 10(-8) M GABA in the washed, frozen and thawed preparation. Bicuculline inhibited the effect of GABA on 3H-diazepam binding. It was found that about 3 X 10(-5) M GABA was present in the assay medium containing crude synaptosomal fraction. These results suggest endogenous GABA is present in, and influences the results of 3H-diazepam binding assays. Furthermore, it appears that GABA and bicuculline affect 3H-diazepam binding through their binding to the specific GABA binding site.


Effect of serotonin on cyclic AMP level in rat hypothalamus slices during development.

The effect of serotonin (5-HT) on cyclic AMP levels in rat hypothalamic slices was age-dependent. The sensitivity of the cyclic AMP system to 5 X 10(-5) M 5-HT already existed in the foetus at 21 days of pregnancy. It reached a maximum on the 7th postnatal day and then decreased with age. In adult tissue the response was still present and was antagonized by two serotonergic antagonists (methiothepin and metergoline). When compared to the progressive increase of 5-HT level in the hypothalamus the data suggest a different evolution of serotonergic innervation and of 5-HT receptors.


Inhibition of fundic strips from guinea-pig stomach: the effect of theophylline on the membrane potential, muscle contraction and ion fluxes.

The effect of theophylline on the smooth muscle cells of the fundic part of the stomach of the guinea pig was investigated. Theophylline hyperpolarized the membrane, inhibited spike discharges and slow waves and produced relaxation of the muscle cells. Furthermore, the theophylline-induced relaxation was not affected in low sodium solution or during inhibition of the sodium pump. Partial inhibition of the theophylline relaxation was seen in muscle depolarized by a high potassium solution and also when ATP was the relaxant. The potassium efflux was enhanced by ATP, but was not markedly changed by theophylline. A change in calcium efflux in the presence of theophylline could not be measured in quiescent preparations. The calcium influx was not changed by theophylline in Krebs, by low Ca, or by high K solution. Furthermore, the tissue content of cyclic AMP was increased by 59.6% in the presence of theophylline (2 x 10(-3) M). The conclusion is reached that the most likely mode of action of theophylline is a suppression of spontaneous activity and an extensive calcium binding to internal sites.


[Seromonitoring of laboratory mouse and rat colonies for common murine pathogens (author's transl)].

During a period from 1973 to 1978, 392 and 225 lots including 12,232 mouse and 8,044 rat individual sera, respectively, were examined for antibodies to murine hepatitis virus, Sendai virus, Bordetella bronchiseptica, Mycoplasma pulmonis, Tyzzer agents, Salmonella typhimurium and Corynebacterium kutscheri. Of mouse lots 94.5% and 39.3% from breeder and user colonies, respectively, were negative for all antibodies examined as well as 31.6% and 17.2% of rat breeder and user colonies, respectively. Among positive lots from mouse users, high positivity rates were seen with Senai virus (47.6%), M. pulmonis (19.0%), and murine hepatitis virus (JHM : 18.2%, MHV : 31.0%), while the rates were high in rat user lots with Sendai virus (24.4%), B. bronchiseptica (39.3%) M. pulmonis (12.5%), murine coronaviruses (JHM : 19.0%, MHV-2 : 28.0%) and tyzzer agents (MSK : 19.6%, RT : 17.9%). These pathogenes with high positivities should be monitored indispensably as a quality control of laboratory mice and rats.


Factors promoting colony stimulating activity (CSA) production in macrophages and epithelial cells.

The regulation of colony stimulating activity (CSA) release from CSA producing cells is a poorly understood process. Using freshly isolated mouse peritoneal cells and a continuous line of mouse thymic epithelial cells a precise and reproducible method of short term culture was developed to study this phenomenon. Serum, endotoxin, lithium, and cyclic GMP stimulated CSA release from both cell types. Particles such as zymosan, inulin, latex, and iron filings stimulated CSA release from mouse peritoneal cells but not from thymic epthelial cells. Theophylline and cyclic AMP inhibited CSA release from both cell types. Trypsin activated guinea pig C3 and C5 did not stimulate CSA release. Cycloheximide, an inhibitor of protein synthesis, completely inhibited CSA release. We believe these findings may reflect mechanisms of in vivo regulation of CSA release.


Cyclic AMP mediated activity stimulation by metronidazole in rat liver.

In the homogenate of the liver biochemically examined 45 hrs after a single 250 mg dose of metronidazole administration, a 45% increase of c-AMP level was found as compared with controls. A marked proliferation of hepatic smooth endoplasmic reticulum was observed. In the light microscope numerous mitotic figures could be seen. No mitotic figures were present in the liver tissue of either treated or untreated animals examined 24, 72 and 120 hrs post administration of metronidazole. Two points seem of interest in the biologic properties of metronidazole: one is the participation of hepatic smooth endoplasmic reticulum in its metabolism, the other--the formation of active metabolites which may posses carcinogenous properties.


Rat endo- and mesothelium lack Fc receptors.

The present study is concerned with the interaction of rat endo- and mesothelium with homologous erythrocytes under various conditions of pretreatment and incubation. Its findings show rat endo- and mesothelial cells a) nonadhesive to native or pretreated erythrocytes irrespective of the presence of gamma-globulin in the medium, b) devoid of primary or cryptic receptors sensitive to the Fc segment of the IgG molecule, and c) provided with binding sites at the oxidized glycocalyx which together with receptor groups of modified erythrocytes share the same class of IgG.


Effect of sodium butyrate in combination with X-irradiation, chemotherapeutic and cyclic AMP stimulating agents on neuroblastoma cells in culture.

Sodium butyrate, X-irradiation, chemotherapeutic agents and cyclic AMP-stimulating agents cuased reduction in the cell number (due to cell death and reduction in cell division) when added individually to mouse neuroblastoma cells in culture. However, the combination of sodium butyrate with X-irradiation, chemotherapeutic and cyclic AMP-stimulating agents produced a greater reduction in the cell number than that produced by the individual agents.


Effects of cadmium on the immune system of mice.

Chronic oral exposure of mice to Cd++ inhibits cell-mediated immunity of delayed type hypersensitivity induced by sheep red blood cells (SRBC). No effect was detected on humoral immune response to SRBC. Spleen cells derived from mice exposed to Cd++ showed in vitro enhanced response to T and B cell mitogens. These results demonstrate that Cd++ exposure alters the immune system of mice.


Regional and strain variation in brain 3':5' cyclic adenosine monophosphate of inbred mice.

Regional variations were found in cAMP levels in flash frozen mouse brains with the pons and cerebellum having higher levels than the cerebrum. There were also strain variations with CBA/J and BALB/cJ having higher levels than C57B1/6J in the pons and cerebellum.


Possible mechanism of rubidium-induced hyperactivity in the rat.

This study suggests that replacement of intracellular potassium by rubidium ions might lower the resting membrane potential. Thus rubidium-treated rats were more responsive to depolarizing influences and generated more cyclic AMP in the brainstem and consequently the behavioral changes.


[Homoanalogues of pyrimidine nucleosides. XII. 3,6-anhydro-1,2-bis-desoxy-1-(pyrimidin- and 5-halopyrimidin-1-yl)-D-arabinohexitols; 3,6-anhydro-1-desoxy-1(5-halopyrimidin-1-yl)-D-glucitols].

Lithium aluminium hydride reduction of 3,6-anhydro-1-o-benzoyl-4,5-o-isopropylidene-2-o-p-tolylsulfonyl-D-mannitol (II) gives with 74% yield 3,6-anhydro-2-deoxy-4,5-o-isopropylidene-D-arabino-hexitol (III). This compound is used as its corresponding 1-o-p-toluene-sulfonate (IV) for the N-1 alkylation of uracil, 5-fluorouracil, thymine and N-acetylcytosine. Acidic cleavage of the acetal protecting group gives the expected bis-homoanalogues of the pyrimidine- and halopyrimidine-nucleosides (V-VIII) which are characterized mainly through their mass spectra and U.V. absorption. 3,6-Anhydro-1,2-dideoxy-1-(5-bromo- and 5-iodouracil-1-yl)-D-arabino-hexitols (XIII, XIV), 3,6-anhydro-1-deoxy-1-(5-bromo- and 5-iodouracil-1-yl)-D-glucitols (XV, XVI) are similarly prepared by direct halogenation of the corresponding uracil derivatives. Results of cytotoxic, cytostatic and antiviral tests are described.


[Effect of preparations that alter the cyclic nucleotide level on acetylation processes].

The action of stimulants and imitators of the adenylate cyclase function (novodrin, sodium fluoride, cAMP), imitators of the guanylcyclase function (cGMP), inhibitors of adenylate cyclase (propranolol), izanylate cyclase (atropin) or phosphodiesterase (euphyllin, papaverin) on the body acetylating capacity measured from acetyl-PABA release, and CoA content in the liver was studied in experiments on male rats. The substances that mainly produce an increase in cAMP in the tissues augment the CoA endogenic pool and, as a rule, stimulate the acetylating capacity of the body. As compared to cAMP, the eczogenic cGMP raises, but to a significantly less measure, the CoA content and does not change the rate of arylamine acetylation. Paraverin is an exception to the rule since it seems to exert a direct action on the enzymatic system of acetylation. When administered to rats cGMP elicits a cAMP-like but a weaker action on the CoA pool. It is suggested that combined use of test substances prolonging cAMP effects and drugs belonging to a group of arylamines is not advisable because of active inactivation of the latter compounds.


Trials with xanthine derivatives in systemic treatment of psoriasis.

The psoriatic epidermis is more active metabolically and its rate of proliferation is faster, showing incomplete differentiation as a result of the imbalance between the low level of cyclic AMP and the high level of cyclic GMP. The low cyclic AMP levels are due to hyperactivity of the phosphodiesterase which hydrolyzes cyclic AMP. Xanthine derivatives are found to be potent phosphodiesterase inhibitors. 15 psoriatic patients were treated with tablets of xanthine derivatives (dyphylline, aminophylline). The obtained results are discussed.


[On neurogenic granular cells and their relationships to the so-called granular cell myoblastoma (author's transl)].

A report on granular cells found in biopsy material is presented. Morphologically identical patterns were seen in a pigmentated granular 'rankenneuroma' and in a patient with a morphea of the tongue. The independent development of granular cells in tumorous and nontumorous tissue supports the idea that the so-called granular cell myoblastoma originates from degenerative alterations of Schwann cells.


[Plasma cortisol response to intravenous beta 1-24 corticotropin as related to the fat mass and its topographic localisation (author's transl)].

Plasma cortisol response induced by 2.5 microgram/kg beta 1-24 corticotropin (1 hour infusion) in obese females was not related to fat mass. The response was increasingly pronounced as fat predominated in the upper body segment, i.e. android obesity. This phenomenon may be interpreted as indicating an increased adrenal capacity to secrete cortisol in such obese patients. Low level but longstanding excess cortisol secretion is probably the cause of the android features of obesity.


[Mechanisms of spontaneous hypoglycaemia in the adult (author's transl)].

Hypoglycaemia increases hepatic glucose output; insulin release is suppressed and the secretion of counter regulatory hormones enhanced. Catecholamines and glucagon seem to play a major role. The brain energy content is initially preserved, but the neuronal activity exhibits a 40-60 % decrease. Neither cerebral blood flow, nor oxygen consumption are altered. In addition to glucose, other substrates are metabolized. Cerebral edema may occur. An insulin-storage defect seems to be the main abnormality in insulinoma beta cell function. The most accurate biological tests are the insulin/glucose ratio, stimulation tests and suppression tests such as fasting and insulin-induced hypoglycaemia. Ectopic release of ACTH, HCG, HLP, glucagon or gastrin, is observed in some malignant insulinomas. When inconclusive, classic localising procedures may be effected by selective venous-blood sampling. Hypoglycaemia of extra-pancreatic tumors results from glucose hyperconsumption and decreases in glucose hepatic output, lipolysis and ketogenesis, related to secretion of insulin-like peptides NSILAs or NSILAp. Rare cases of hypoglycaemia related to insulin auto-antibodies of unknown origin have been reported. Alcoholic hypoglycemia results from diminished hepatic glycogen content, alcohol dehydrogenase pathway blockade, reduction of gluconeogenesis defect in the alcohol catabolic catalase pathway and enhancement of peripheral glucose consumption.


Low density lipoprotein receptor activity in fibroblasts cultured from diabetic donors.

Low density lipoprotein (LDL) receptor activity was evaluated in cultured skin fibroblasts from diabetics and nondiabetic controls to evaluate whether intrinsic abnormalities of the LDL pathway exist, which might account for the premature atherosclerosis associated with diabetes mellitus. LDL receptors did not differ between cells grown from 16 diabetics (7 insulin-dependent, 9 non-insulin-dependent ) or from 16 nondiabetic controls. An inverse relationship between LDL receptor activity and cell density was observed (y = 1.35x-1.22, r = 0.90, P less than 0.001), which appeared the same for diabetic and nondiabetic cells. Normalized values for LDL degradation by diabetic and nondiabetic cell strains were 1.52 +/- 0.42% of added LDL/10(6) cells and 1.34 +/- 0.28, respectively (P = NS). The kinetics of the LDL receptor also appeared to be the same in cells derived from a diabetic and a nondiabetic donor. LDL receptor activity in diabetic cells increased appropriately in response to physiologic concentrations of insulin in the incubation mediu. Thus, LDL receptor activity appears to be normal in diabetic cell strains. Therefore, these results do not support the possibility that alterations in the LDL pathway contribute to the accelerated atherosclerosis associated with diabetes.


The prestalk-prespore pattern in cellular slime molds.

In cellular slime molds the slugs become divided into two regions with different properties, and anterior prestalk-zone and a posterior prespore zone. Although the cells in these zones are normally destined to form the stalk cells and spores of the fruiting body, respectively, they are not irreversibly committed to one sort of differentiation or the other during the slug stage. The volume ratio of the two zones remains almost constant over a wide range of slug sizes. If the prestalk-prespore pattern is distrubed by removing tissue from the slug, conversion of tissue from prestalk to prespore or vice versa occurs as necessary to restore a normal pattern with normal proportions. Conversions also occur in both directions during normal development. The initial formation of the prestalk-prespore pattern may well involve sorting-out, but other mechanisms must be invoked to account for regulation. We describe three different models of the generation of the prestalk-prespore pattern, the'cell-contact model' of McMahon, in which pattern is created by interactions of cells with their immediate neighbors, the 'positional-information model' of various authors, in which pattern formation involves an overall gradient and a gradient-reading mechanism, and the 'activator-inhibitor model' of Gierer and Meinhardt, in which the prestalk-prespore pattern is formed by a system of diffusible substances that affect one another's production. The activator-inhibitor model is the most successful of the models at describing the known features of the prestalk-prespore pattern. The various models lead to a number of distinctive predictions. According to the cell-contact model, small transplants may cause gross changes in the prestalk-prespore pattern, and mutants may exist which severely disrupt pattern formation even if diluted with a large excess of wild-type cells. Positional-information models predict the existence of 'gradient-reading mutants'; slugs that are a mixture of such mutants and wild-type cells would show two prestalk-prespore boundaries, one at the mutant and one at the normal position. Both the activator-inhibitor model and some versions of the positional-information model predict that small transplants will sometimes induce accessory prestalk or prespore zones; the quantitative characteristics of these effects may allow one to make a case in favor of one or other of the two models. Finally, the activator-inhibitor model leads one to expect that mutants may be isolated which normally show accessory prestalk or prespore zones. A search for these phenomena may help determine whether the activator-inhibitor model will continue to enjoy its present preeminent position.


Immunohistochemical localization of cyclic GMP in aggregating Polysphondylium violaceum.

Using indirect immunofluorescence technique, it has been possible to localize cyclic GMP in Polysphondylium violaceum cells. The bound cyclic nucleotide is localized throughout the cell during early stages, however, this staining increases and there is marked localization of cyclic GMP in the nuclear areas of the cells when aggregation is in full swing. Over 90% of the cells exhibited intense nuclear staining by 6 h and this decreased to less than 10% by 10 h.


Persistence of phage lambda DNA in genomes of mouse cells transformed by lambda-carrying SV40 vectors.

To test the suitability of simian virus 40 (SV40) DNA as a vector for inserting DNA segments into the chromosomes of mammalian cells, an EcoRI-A fragment of bacteriophage lambda DNA was covalently joined to a fragment of SV40 DNA and used to transform mouse cells in culture. Three independent, morphologically transformed clones were obtained that were positive for SV40 T-antigen by immunofluorescence staining. DNA from each transformant was examined by restriction enzyme analysis and found to contain both lambda and SV40 sequences. Co-migration of some fragments containing lambda and SV40 sequences following digestion of transformed cell DNA by each of four different restriction enzymes indicated that part of the retained lambda and SV40 DNA was linked in two of the three lines. In the third line, however, none of the restriction fragments had both lambda and SV40 sequences. Although the presence of non-integrated lambda DNA was not excluded, at least some of the lambda DNA appeared to be linked to host cell DNA. Results of digestion by EcoRI suggested that in some cases the transforming linear molecule had probably circularized prior to integration.


An autopsy case of progressive dementia showing special intraplasmic inclusions in diffuse cerebral cortex.

A 29-year-old male characterized by progressive dementia has been described. Histopathologically, the changes obtained were as follows: 1) Some atrophic nerve cells contained homogeneous round or oval intraplasmic inclusions which could be stained with periodic acid-Schiff and coupled tetrazonium. From these examinations the inclusions consisted of neutral mucopolysaccharide including protein. 2) Mild neuronal loss was diffusely present. 3) Most of the remaining nerve cells were atrophic. The above-described findings were observed diffusely in the whole cortex of the brain. To our knowledge, the present case does not belong to the category of any well-known pathological diseases.


Percutaneous transhepatic portography in bile duct carcinoma. Correlation with percutaneous transhepatic cholangiography and angiography.

In 16 patients with carcinoma of the extrahepatic bile ducts, percutaneous transhepatic portography (PTP) and cholangiography (PTC) were performed. Fourteen of these patients also had angiography, which failed to show the tumor in 7 cases. Findings indicating non-extirpability of the tumor were demonstrated by angiography in 3 patients. At PTP non-extirpability was confirmed in these cases. PTP further indicated non-extirpability in one patient and gave hints of infiltration of the liver by the tumor in 5 patients because of intra or extrahepatic invasion of the portal vein.


[The radiologic diagnosis of inguinal hernias. Experience with the use of intraperitoneal iodinated contrast media (author's transl)].

By injecting an iodine contrast medium into the abdominal cavity and by placing the patient in a position with the table somewhat elevated, it is easy to visualize the inguinal region. The examination is performed with the X-ray tube angulated caudally, and exposures are made while the patient is straining. The method is well adapted for patients with obscure pains in the groins, for patients operated for hernias but not cured of pain and for patients in whom symptoms recur postoperatively without clinical evidence of hernial relapse. The value of the methode appears from four short case reports.


Plasma lipids and high density lipoproteins during oral contraception with different combinations of ethinyl estradiol and levonorgestrel.

Seventy-five menstruating women seeking contraceptive advice were randomly allocated to treatment with combined oral contraceptives containing either ethinyl estradiol 50 micrograms + levonorgestrel 250 micrograms (50/250), ethinyl estradiol 30 micrograms + levonorgestrel 150 micrograms (30/150) or ethinyl estradiol 50 micrograms + levonorgestrel 125 micrograms (50/125). The concentrations of cholesterol, triglycerides, phospholipids, high density lipoprotein (HDL)-cholesterol and HDL-phospholipids were determined after one, three and six months and compared to the mean of two determinations of the same parameters before medication. Triglycerides increased by 18--42 per cent after 1--6 months of treatment with 50/125. The HDL-cholesterol and HDL-phospholipids were reduced by 10 per cent during 50/250 treatment. No other parameters showed any consistent alteration in any of the treatment groups. Raised triglyceride concentration and/or decreased HDL concentration increases the risk for cardiovascular disease. It is therefore suggested that in order not to alter the HDL concentration a combined oral contraceptive agent should not contain more gestagen-androgen than corresponding to 125--150 micrograms of levonorgestrel. To avoid a rise of the triglyceride level the weight relation between levonorgestrel and ethinyl estradiol should be about 5:1.


The effects of growth hormone, thyroxine and insulin on the activities of reduced nicotinamide adenine dinucleotide phosphate dehydrogenase, glucose-6-phosphatase and glycogen phosphorylase in fetal rat liver.

Growth hormone (GH), thyroxine (T4) and insulin were injected, in utero into 20.5 day-old rat fetuses to study the effects of these hormones on the activities of liver NADPH dehydrogenase, glucose-6-phosphatase and glycogen phosphorylase. It was found that at 21.5 days of gestation, GH increases the fetal liver glucose-6-phosphatase activity and decreases the liver glycogen phosphorylase activity. T4 treatment augments the activity of NADPH dehydrogenase even at 0.3% of the dose shown previously to produce premature elevation of activity. Prior to this experiment T4 in large doses has been shown to be capable of elevating glucose-6-phosphatase. However, at the lower T4 dose used, no treatment effect was observed. The fetal rat liver is responsive to insulin at 21.5 days and insulin was able to depress glucose-6-phosphatase activity. Thereby, showing that the influence of insulin on this enzyme begins prior to birth instead of just subsequent to birth.


Congenital hypoaldosteronism. Thirteen year follow-up in identical twins.

Identical male twins suffering from congenital hypoaldosteronism due to a rare adrenal enzyme deficiency between corticosterone and aldosterone were followed-upfrom birth till their present age of 13 years. The symptoms of salt loss disappeared and normal growth rate resumed following treatment with DOCA and salt supplementation. Discontinuation of mineralocorticoid administration at the age of 7 years resulted during a 5-year period in a marked decline in their growth rate. Labororatory data revealed a persistent, albeit less pronounced, metabolic impairment. Mineralocorticoid administration was resumed and the twins entered normal puberty and increased their growth rate, emphasizing their need for continued mineralocorticoid administration to maintain adequate growth rate and development.


Reduced response of plasma aldosterone to acute ACTH stimulation during long-term treatment with spironolactone in essential hypertension.

The response of plasma aldosterone (PA) to ACTH administration (250 micrograms alpha 1-24 ACTH i.m.) before and during treatment with spironolactone (Sp, 75--100 mg/day) for at least 8 months was studied in 11 patients with essential hypertension. These responses were compared with those before and during prolonged treatment with hydrochlorothiazide (Th, 50--75 mg/day), with or without potassium supplement, in 14 hypertensives. PA and plasma cortisol (PC) were determined by radioimmunoassay in which Sp showed minimal cross-reactivity. Both Sp and Th treatments caused similar increases in plasma renin activity accompanied by nearly identical decreases in blood pressure and body weight. PA was also increased by both treatments, but to a significantly greater extent in the Sp-treated group. Serum potassium concentration was increased only by Sp treatment. The response of PA, but not of PC, to acute ACTH stimulation was blunted in the Sp-treated group. That is, the maximal increment of PA above the baseline level was significantly lower during Sp treatment than either before Sp treatment of during Th treatment. These results demonstrate that long-term treatment with Sp can inhibit aldosterone production by acute ACTH stimulation in patients with essential hypertension.


Studies on the mode of action of gonadotrophin-inhibiting material (anti-LH) isolated from human urine.

Gonadotrophin-inhibiting material (GIM) was able to inhibit the binding of 125I-hCG to rat Leydig cells, suggesting that its inhibiting properties are due to its ability to complete for the hCG binding sites on Leydig cells. Binding of fluorescein isothiocyanate-tagged GIM to Leydig cells was also seen. The effect of GIM on hCG-induced adenylate cyclase activation and testosterone production was also studied. It was found that there was a dose-dependent inhibition of both these effects.


Characterization of cells cultured from early lactation milks.

Two major types of cells can be cultured from early lactation human milks: a colony-forming epithelial cell and an adherent nondividing cell referred to as a foam cell. The epithelial cells show a positive reaction with a specific antiserum reactive against membrane components of the milk fat globule, whereas the foam cells do not. The nondividing foam cells are phagocytic and can be killed by silica particles; they produce lysozyme, are resistant to trypsinization, and have Fc receptors. These properties, together with the lack of reaction with antiserum to the milk fat globule membrane, suggest that the foam cells are not terminally differentiated epithelial cells, but tissue macrophages.


Interferon production by mammalian cells grown in a serum-free medium.

Interferon, produced by rabbit heart cells grown in a serum-free medium, failed to protect rabbit heart serum-free cells, but protected rabbit heart serum-containing-medium cells against vaccinia and vesicular stomatitis virus. Interferon produced in serum-free cells had a greater species specificity than that produced in serum-containing media. The difference in activity was shown to be due to lack of adsorption by serum-free-medium cells.


Collagen synthesis in normal BHK cells and temperature-sensitive chemically transformed BHK cells.

Collagen synthesis in normal BHK 21/cl 13 and chemically transformed temperature sensitive BHK 21/cl 13 cells (Me2N4) was assessed by examination of hydroxyproline formation and collagenase-susceptible protein. The Me2N4 cells lost their ability to synthesize collagen at both permissive and nonpermissive temperatures for transformation. These conclusions were confirmed by polyacrylamide-gel eletrophoresis and CM-cellulose chromatography. Prolyl hydroxylase activity was present in both normal and transformed cells even when no collagen could be demonstrated. The production of noncollagen protein, although decreased in the transformed cell, did not change as drastically as the collagen synthesis.


Regulation of bradykinin-induced cyclic amp response by quinacrine and prostaglandin E2 and F2 alpha in human synovial fibroblasts.

Bradykinin induces an increment in intracellular cyclic AMP concentrations of human synovial fibroblasts and evokes the release of [3H]arachidonic acid and [3H]-E prostaglandins from human synovial fibroblasts pre-labeled in their phospholipids. Both these bradykinin-induced reactions are inhibited by quinacrine, an inhibitor of phospholipase A activity. The cyclic AMP response of human synovial fibroblasts to bradykinin is potentiated by prostaglandin E2 and inhibited by prostaglandin F2 alpha. These data emphasize the critical role of the prostaglandin system in reactions induced by bradykinin and suggest mechansims by which inflammatory reactions due to bradykinin may be modulated.


Unique characteristics of superoxide production by human eosinophils in eosinophilic states.

Eosinophils from patients with peripheral blood eosinophilia and human neutrophils from normal subjects and patients with neutrophilia produced superoxide anion (O2-) in vitro at similar rates in the absence of stimulation and exhibited comparably increased rates of O2- production during the initial 1 h of incubation with opsonized zymosan. In the presence of opsonized zymosan, the rate of O2- production by eosinophils was constantly high for 3 h, whereas the rate of production by neutrophils fell by more than 65% after 1 h. Consequently, the amount of superoxide produced by phagocytizing leukocytes was twofold higher for eosinophils than for neutrophils at 3 h. O2- production by cell-free sonicates of zymosan-stimulated eosinophils and neutrophils exhibited the same preference for NADPH over NADH. One mM sodium azide significantly decreased the generation of O2- by phagocytizing eosinophils, but lacked an effect on neutrophils. the prolonged release of O2- by eosinophils engaged in phagocytosis may contribute both to their unique microbicidal profile and to the capacity of eosinophils to injure host tissues in some eosinophilic syndromes.


Tobacco smoke. Effects on pulmonary host defense.

Tobacco smoke affected both the metabolism and function of pulmonary alveolar macrophages (PAM). Phagocytosis of viable Staphylococcus aureus and inert starch particles was minimally but consistently depressed in PAM from rats exposed to tobacco smoke for six months. Oxygen consumption, superoxide and hydrogen peroxide release, and hexose monophosphate shunt activity were elevated in cells from smokers. Oxidation of glucose, labelled in the carbon-six position, remained unchanged. All observed effects of tobacco smoke on oxygen metabolism occurred during phagocytosis and did not affect the basal metabolism of the nonstimulated cell.


Effect of estrogens on the myeloperoxidase-mediated antimicrobial system.

Estradiol 17 beta prevented the fall in the microbicidal activity of the myeloperoxidase-H2O2-halide system induced by high H2O2 concentrations. In contrast, when the H2O2 (and halide) concentrations were low the myeloperoxidase-H2O2-halide antimicrobial system was inhibited by estradiol. These properties of estradiol 17 beta were shared by estradiol 17 alpha, estrone, estriol, ethinyl estradiol, and phenol, but not by estradiol-3-benzoate, testosterone, progesterone, hydroxyprogesterone, cortisone, hydrocortisone, or deoxycorticosterone.


Cell-mediated immunity to varicella-zoster virus measured by virus inactivation: mechanism and blocking of the reaction by specific antibody.

The process whereby varicella-zoster (V-Z) virus is inactivated in vitro by immune human peripheral blood leukocytes stimulated with V-Z antigen was examined. It was found that stimulation of leukocytes by V-Z antigen, but not by other viral antigens, was required for inactivation of V-Z virus to occur. Viral inactivation could be blocked by addition of V-Z antiserum to either the stimulation phase of the reaction or the inactivation phase, further demonstrating the specificity of the reaction. In addition these blocking experiments suggested that modulation of V-Z membrane antigen by antiserum occurred with an accompanying loss of immunological recognition of virus-infected cells. Inactivation of V-Z virus in vitro in this study appeared not to be dependent upon the secretion of interferon or upon antibody-dependent cellular cytotoxicity. The specific cells required for V-Z inactivation were T lymphocytes and monocytes (macrophage precursors).


Cellular and humoral immune responses to varicella-zoster virus in immunocompromised patients during and after varicella-zoster infections.

Humoral and cell-mediated immune responses to varicella-zoster (V-Z) virus were assessed in patients during and after V-Z infections. Ongoing V-Z infections was associated with minimal cellular immunity but not necessarily with poor humoral immunity. Recovery from V-Z infection was associated with a vigorous cellular immune response. Cell-mediated immunity to V-Z virus was demonstrable for years after varicella, but responses were lower in immunocompromised patients than in normal individuals.


Inhibition by cholera toxin of rat polymorphonuclear leukocyte chemotaxis demonstrated in vitro and in vivo.

The effect of cholera toxin on the chemotaxis of rat polymorphonuclear leukocytes (PMN) was studied using a technique in which the movement of the cells towards a laser-lysed erythrocyte is followed under a phase-contrast microscrope. In vitro studies indicated that the intact toxin was capable of inhibiting PMN chemotaxis in a dose-dependent manner at doses ranging from 1 to 100 ng/ml. Subunits A and B of the toxin were without inhibitory activity when used alone, but after recombination their ability to inhibit chemotaxis was similar to that of the intact toxin, suggesting that the toxin is acting intracellularly. Cholera toxin has been reported to act in other systems via stimulation of adenyl cyclase with consequent elevation of intracellular cyclic adenosine 5'-monophosphate (cAMP) levels. It appears that this mechanism may also account for its ability to inhibit chemotaxis since there was a correlation, at all doses tested, between inhibition of chemotaxis and increased intracellular cAMP levels. Cholera toxin was also found to be active in vivo in that, after intrapleural injection of the toxin, the chemotaxis of cells subsequently recovered from the pleural cavity was markedly reduced. These results support previous findings which suggest that modification of leukocyte cAMP levels can influence the chemotactic responsiveness of these cells.


Antibody responses and interferon titers in the respiratory tracts of mice after aerosolized exposure to influenza virus.

We studied the temporal appearance of immunoglobulins (immunoglobulins G1, G2, M, and A) and interferon in lung lavage fluids of mice after aerosol exposure to influenza virus in six animal groups in which mortality rates ranged from 0 to 24%. Immunoglobulin levels in the lung lavage fluids were markedly higher in mouse groups with higher mortality rates (16, 20, and 24%) than in those with low mortality rates (0, 2.5, and 7.5%). Analysis of serum albumin in the respiratory secretions as an index of edema indicated that increased immunoglobulin G levels during the early phase of infection were due to increased vascular permeability. The detection of virus-neutralizing antibodies and antibodies reactive with influenza virus antigens in the lavage fluids at 6 to 8 days postinfection suggested local immunoglobulin synthesis as a result of antigenic stimulation. Both systemic and local antibody productions contributed to immunoglobulin levels in the respiratory secretions after aerosolized influenza virus infection. Peak levels of interferon in the lavage fluids were reached before detection of significant levels of virus-neutralizing antibody in the serum or the lung lavage.


Replication of vesicular stomatitis virus facilitated by Shope fibroma virus in vivo.

Previous studies show that Shope fibroma virus facilitates replication of vesicular stomatitis virus (VSV) in some rabbit cells grown in vitro. In the present investigation, the possibility that these two viruses can also interact in vivo was determined. Rabbits inoculated intradermally with both viruses together, or each separately, were examined for the formation of lesions or tumors and for the production of infectious virus. The presence of VSV interfered with tumorigenesis by Shope fibroma virus. In tumors already formed, production of infectious VSV was greater than in normal skin. Hence, each virus affected the other. Sera and tissues of normal rabbits were found to contain a substance which inhibits VSV; this may act to limit replication of VSV in rabbit skin. In addition, cultured rabbit skin cells appeared to adsorb VSV inefficiently. When persistently infected by Shope fibroma virus, however, adsorption of VSV was markedly improved. Our results suggest that in vivo Shope fibroma virus may facilitate adsorption of VSV to reduce the effect of a natural inhibitor and consequently enhance production of infectious virus.


Metabolic and functional characteristics of alveolar macrophages recovered from rats exposed to marijuana smoke.

Pulmonary alveolar macrophages were obtained by bronchopulmonary lavage from male rats after 30 consecutive days of in vivo exposure to marijuana and tobacco smoke. No significant differences were found between either group of experimental animals and controls in the number of cells recovered, the protein content per 10(6) cells, or the percentage of cells that adhered to plastic surfaces. The ability of macrophages to phagocytize viable bacteria was not affected by exposure to either marijuana or tobacco smoke in that both treatment groups ingested Staphylococcus aureus over a 60-min period as well as did control cells. Differences were found between the groups, however, with respect to cellular metabolism. Marijuana smoke inhalation caused a small decrease in the amount of oxygen consumed by macrophages during phagocytosis, as compared with control cells. This may have been reflected in the even greater decrease in superoxide formation observed during particle engulfment by these treated cells. Tobacco smoke, on the other hand, increased oxygen consumption and was without effect on superoxide release. Neither tobacco nor marijuana smoke treatment had an effect on the direct oxidation of glucose via the hexose monophosphate shunt. Our results indicate that, despite several metabolic alterations in response to marijuana and tobacco smoke, alveolar macrophages were not compromised with respect to their ability to ingest a particulate challenge.


Cytokine production by blue tongue virus-infected fetal sheep cells.

The migration inhibition of guinea pig peritoneal macrophages by a factor(s) from media obtained from blue tongue virus-infected monolayer cultures was studied. Medium from blue tongue virus-infected sheep fetal cell cultures inhibited migration of guinea pig macrophages from agarose droplets. Medium from control cultures and stock virus did not inhibit macrophage migration. Medium containing migration inhibiting factor(s) in vitro induced an inflammatory reaction in the skin of a newborn sheep. The inflammatory reaction was observed 20 h after intradermal inoculation. The skin reaction consisted of infiltrates of mononuclear leukocytes in the superficial dermis. Control medium and stock virus caused no skin reaction.


Enhancement of granulocyte chemiluminescence with hydroxyl radical scavengers.

Addition of hydroxyl radical scavengers (benzoate, ethanol, and mannitol) to granulocyte chemiluminescent reactions significantly enhanced light generation without altering granulocyte oxygen consumption or superoxide production.


Induction of virus-neutralizing antibody by glycoproteins isolated from chicken cells infected with a herpesvirus of turkeys.

The production of virus-induced proteins in chicken embryo fibroblast cells infected with a herpesvirus of turkeys was studied. It was found that glycoproteins isolated from membrane-rich fractions of infected cells by affinity chromatography using concanavalin A induced neutralizing and precipitating antibody in rabbits and chickens. After analytical electrophoresis, such isolates were found to contain three polypeptide bands of between 100 x 10(3) and 120 x 10(3) molecular weight not present in glycoprotein extracts of uninfected cells, and these polypeptides were further purified by preparative polyacrylamide gel electrophoresis. Inoculation of chickens with purified material also resulted in the production of precipitating and neutralizing antibody, showing that these high-molecular-weight polypeptides play a role in the humoral immunity to Marek's disease. Challenge of these chickens with virulent Marek's disease virus revealed that a partial protection was afforded by the inoculated glycoproteins.


Serological response of the bovine fetus to bovine viral diarrhea virus.

Susceptible pregnant heifers were inoculated with bovine viral diarrhea virus at 150 days of gestation and earlier. Fetuses were surgically collected at selected times after inoculation. Serum immunoglobulins were quantitated, and the presence of specific antibodies was determined. In fetuses from heifers inoculated at 150 days, immunoglobulin M (IgM) appeared approximately 2 weeks after inoculation and was followed in 7 days by IgG1. Later IgG2 was detected in the sera of three fetuses. Serum-neutralizing and complement-fixing antibodies were first detected in a fetus taken at 206 days of gestation. Fetuses taken at later times also had specific serum antibodies. Possible explanations for the appearance of serum immunoglobulin substantially before specific bovine viral diarrhea antibodies include the viral alteration of host tissues rendering them antigenic viral activation of polyclonal B cells, and viral modulation of virus-specific lymphocytes causing specific interference with the appearance of antiviral antibodies. In one of the fetuses having IgG2, the serum also contained IgA. Placental leakage of material immunoglobulins was thought to be responsible for the presence of IgA and IgG2 in this fetus. Small quantities of IgM were found in the serum of two fetuses taken from heifers inoculated between 65 and 95 days of gestation, but specific antibodies were found in none.


Elevated titer of antibodies to Simian sarcoma virus envelope antigen (gp70) and normal response to influenza virus in untreated Danish Hodgkin's patients.

Untreated Danish Hodgkin's disease (HD) patients and paired age- and sex-matched controls were tested for serum antibodies to Epstein-Barr virus (EBV), six strains of influenza virus and Simian sarcoma virus/Simian sarcoma-associated virus [SSV(SSAV)] antigens. HD sera showed significantly elevated titers against EBV and both increased incidence and mean titer of antibodies to the envelope glycoprotein of SSV(SSAV), whereas testing against the influenza viruses revealed no differences between HD and controls. A focus reduction assay demonstrated a low incidence in HD and controls of sera with neutralizing effects against SSV(SSAV) and the "baboon" C-type virus component of the human HL-23 virus complex, which seemed coupled to HL-A type W19.


Growth of diploid, Epstein-Barr virus-carrying human lymphoblastoid cell lines heterotransplanted into nude mice under immunologically privileged conditions.

Human Epstein-Barr virus-carrying lymphoid cell lines which have been classified on the basis of studies on clonality and morphological, chromosomal and functional parameters as lymphoblastoid cell lines (LCL) of presumed non-neoplastic origin were inoculated intracerebrally into nude mice. All eighteen of them grew, killing the host mice within 7 to 25 days, except for 2 which grew more slowly. At autopsy, the brain of the nudes was found to be invaded by infiltrating lymphomas. Sixteen of these lymphomas, when recultured in vitro, gave rise to cell lines with growth properties and morphology indistinguishable from those of the inoculated LCL. Chromosomal examinations showed that 3/7 cell lines injected, which grew as lymphomas in the brain, were still normal diploid on reexplantation whereas the remaining four had become aneuploid. Four lines derived from intracerebral lymphomas (2 diploid, 1 aneuploid and 1 untested) were inoculated subcutaneously into adult nude mice. None of them grew. When the corresponding four original LCL lines were inoculated subcutaneously into newborn nude mice, they grew rapidly, but failed to do so in newborn normal mice or intracerebrally in adult normal mice. One such line, U-1450, was treated with anti-lymphocyte serum (ALS). Small nodules developed at the site of inoculation. From one nodule a cell line was cultured, 1450 ALSAD. It was morphologically indistinguishable from the line of origin. The lines obtained from nude mice inoculated with polyclonal LCL seem to have a restricted clonal representation, but were not monoclonal, as evidenced by analyses of their pattern of immunoglobulin synthesis.


Vascular basophilia in ocular and orbital tumors.

The occurrence of vascular basophilia in ocular tumors has been a selective histologic feature of retinoblastomas. We recently observed a metastatic oat-cell carcinoma to the choroid which also demonstrated such a vascular hematoxyphilia. Histologic review of a variety of ocular and orbital metastatic carcinomas failed to yield a similar basophilic pattern. Examination of 100 consecutive retinoblastomas for vascular basophilia revealed an incidence of 6.0%. Similar material was not seen in any of 125 melanomas, including 10 with areas of necrosis. Histochemical studies showed the basophilic material to be DNA, and electron microscopy revealed the nuclear debris of pyknotic tumor cells to be continuous with identical material surrounding the adjacent blood vessels. The pathogenesis of vascular deposition of DNA in these two ocular tumors remains unclear. This finding most likely represents a form of tumor activity requiring comparatively healthy blood vessels to adequately precipitate liberated nucleic acids being filtered from the necrotic and degenerating tumor tissue.


Adenovirus types 8 and 19 infection of rabbit corneal organ cultures.

Adenovirus types 8 and 19 are responsible for a large number of cases of acute follicular conjunctivitis, mainly in the form of epidemic keratoconjunctivitis (EKC). This study successfully demonstrates the establishment of adenovirus infection in rabbit corneal organ cultures. Normal rabbit corneas were maintained in culture without significant morphologic alterations for up to 7 weeks. Adenovirus types 8 and 19 replication in rabbit corneas was demonstrated by viral growth curves and subpassage studies. Intracellular localization of adenoviral antigens was confirmed with immunofluorescent techniques. The characteristic cytopathic effects of adenoviral infection were observed with the use of various histochemical stains. Thus an in vitro corneal organ culture model was established for the study of the pathogenesis of corneal lesions in adenovirus EKC.


Scanning immunoelectron microscopy markers.

Ultrastructural analysis of the cell surface and of membrane components such as antigens or receptor sites by scanning immunoelectron microscopy (immuno-SEM) has been the subject of extensive investigation during the past few years. We review the various immunologic and cytochemical techniques applied to SEM, which have employed latex particles, viruses, bacteriophages, protein A, ferritin, gold granules, hemocyanin and peroxidase as markers, and the advantages and disadvantages of these techniques. From current data, it is clear that immuno-SEM has much to offer in determining the distribution of specific cell surface sites and in positive and unambiguous identification of cell types in heterogeneous cell populations.


Tantalum oxide, silica and latex: effects on alveolar macrophage viability and lysozyme release.

Tantalum an experimental bronchographic material, may be retained in the lungs for a prolonged period following bronchography. The alveolar macrophage (AM) is a cell with potential for clearing tantalum particles from the airways. We studied the in vitro effects of tantalum oxide and two other particles, silca and latex, on rabbit AM viability and lysozyme release over 30 hours. Results indicate: 1) tantalum oxide, silica, and latex particles are ingested by rabbit AM in culture; 2) tantalum oxide and silica are both toxic to AM in vitro; and 3) tantalum oxide exerts its toxic effects less rapidly on AM than does silica. On the basis of these in vitro culture results we conclude that tantalum oxide may be toxic to alveolar macrophages in vivo. Delayed lung clearance of tantalum oxide particles may be due in part to their toxic effects on alveolar macrophages.


[Papillomatous lactic-duct adenoma (pseudoPaget's disease of the mamilla)].

Papillomatosis and adenomatosis of the nipple ducts can be interpreted clinically as Paget's disease. Three patients with subareolar papillomatosis and adenomatosis are described. The name papillary adenoma of the nipple ducts (pseudo-Paget's disease has been proposed. The differentiation between the papillary adenoma of the nipple ducts and the genuine Paget's disease needs histological examination. The differential diagnosis is very important, because local excision of the diseased ducts represents the treatment of choice of the papillary adenoma of the nipple ducts. Radical mastectomy is always necessary in genuine Paget's disease since the adjacent lactiferous ducts are involved by the carcinoma.