The peripheral lymphocyte count as an aid in the clinical staging of lung cancer. The association between disease staging and peripheral lymphocyte count (PLC) was studied in 178 patients with histologically verified lung cancer. Mean PLC varied inversely with the stage of disease (p less than .01). The mean values were 2821, 2217, and 1594 for Stages I, II and III respectively. Thoretical normal distributions were generated about the mean peripheral lymphocyte count for each stage. Relative frequencies of lung cancer staging based on the PLC are estimated utilizing these curves.


Watery diarrhea syndrome. Two unusual cases and further evidence that VIP is a humoral mediator. Two unusual cases of the watery diarrhea syndrome are presented. In one patient an adrenal medullary tumor, a pheochromocytoma that produced vasoactive intestinal polypeptide (VIP) was excised with total relief of symptoms. The second patient a 65-year-old man with abrupt onset of massive watery diarrhea that led to acidosis and coma was symptomatically controlled for one year on 10 mg/day of prednisone. Elevated levels of VIP returned to normal after prednisone therapy was started. A benign islet cell tumor not localized by angiography was removed by distal pancreatic resection. Tissue levels of VIP were markedly elevated. VIP is a humoral mediator of the water diarrhea syndrome. Both benign and malignant pancreatic and extrapancreatic tumors may cause the watery diarrhea syndrome. Steroids may cause symptomatic relief of the diarrhea by lowering peptide levels to normal. The term watery diarrhea syndrome may be more accurate than the pancreatic cholera syndrome.


Reappraisal of solitary bronchiolar (alveolar cell) carcinoma of the lung. Twelve patients with solitary bronchiolar carcinoma had lobectomy and were followed for up to 16 years. The concept of a multicentric origin of bronchiolar carcinoma, maintained for more than eight decades, should be discarded. The neoplasm arises indolently and usually in an area of pulmonary fibrosis. After lobectomy patients can now expect to follow one of four courses: (1) to be alive and well without recurrence; (2) after several years to have pulmonary recurrence or a new carcinoma; (3) with minute spread at the time of lobectomy to have metastasis develop in a short period; or (4) to die of unrelated conditions. The overall 5-year survival with this tumor is about 75%. Late recurrence or the development of another primary tumor, however, prompts the need for prolonged follow-up. Immunologically, patients have circulating antibodies when well and demonstrable circulating antigens with recurrence. The survival rate of selected patients with solitary bronchiolar carcinoma (eliminating those patients with microscopic spread from the primary neoplasm at the time of resection and those dying of other causes) was 100% after 5 years and 75% after 10 years.


Influence of triethylcholine on autonomic transmission in vitro. The influence of triethylcholine (TEC) on isolated autonomic transmission has been investigated in some isolated preparations. The guinea-pig hypogastric nerve-vas deferens-urinary bladder preparation for studying both cholinergic and adrenergic transmission simultaneously is described. Incubation with TEC caused impairment of cholinergic transmission but failed to inhibit adrenergic transmission. Employing TEC as a pharmacological tool, no evidence for a cholinergic link could be obtained in adrenergic transmission in the isolated vas deferens and rabbit ileum preparations.


Endometrial carcinoma following estrogen therapy for breast cancer. Report of three cases. Three patients with metastatic breast cancer responded to diethylstilbestrol (DES) therapy, but later they developed endometrial carcinoma. Evidence is presented to support the hypothesis  that endometrial carcinoma can occur in breast cancer patients receiving diethylstilbestrol therapy.


[Gastritis with hypoproteinemia in children. Relation to Menetrier's disease and cytomegalic inclusion disease]. The case report concerns a 2 years old girl with hypoproteinemia associated with gastritis. The child recovered completely 8 weeks later. Cytomegalovirus was excreted in urine. The authors suggest that such a clinical picture should not be called Menetrier's disease. The role of cytomegalovirus is discussed.


[Adenomegalic cytophagic sinusal histiocytosis. Morphological and immunologic study of a case]. The case of a 14-year-old boy of North-African origin, presenting a phagocytic sinus histiocytosis is reported. The main features of this now classical disease are illustrated: its localization to the neck, its chronicity, the fact that it is well tolerated, the pseudotumoral appearance of the adenomegaly as well as the intensive phagocytic (essentially lymphocytophagic) activity of the sinus macrophages. The bringing to light in a picture of hyperimmunity of an elevated percentage of antibodies against the measles and EB viruses, associated with a temporary depression of the cellular immunity, suggest that this lymphophagocytosis could be controlled and facilitated by preferential opsonization of lymphoid cells carrying on their membrane the antigen of the virus or viruses initially responsible.


[Etiology and associations of pituitary dwarfism. Study of a series of 135 cases]. Among 135 consecutive cases of hypopituitary dwarfism, 42 related to detectable intracranial tumour or defect. In 13 cases the tumour had been previously operated and/or irradiated. In 29 others the defect was suggested by neurological abnormalities or headache, skull radiographs, ocular examination or by associated post-hypophyseal deficiency and was demonstrated by pneumoencephalography. When none of these associated abnormalities was found, pneumoencephalography failed to demonstrate any intracranial lesion. Among the 93 so-called idiopathic cases there was a large majority of males (60/93) with a history of birth difficulties (34/60) and especially of breech delivery (23/60). TSH, FSH/LH and ACTH deficiencies were associated to GH deficiency in 81% of patients with detectable intracranial lesions, 57% of male and 39% of female idiopathic cases. The number of patients with idiopathic isolated GH deficiency was similar in boys and girls, suggesting in them the hypothesis of a recessive autosomic genetic defect in spite of the scarcity of familial cases. Peculiar clinical associations may contribute to the diagnosis.


Necrotizing pancreatitis due to simian adenovirus type 31 in a rhesus monkey. Simian adenovirus type 31 was isolated from pancreatic tissue of an 8-year-old male rhesus monkey that died as the result of acute necrotizing pancreatitis. Histologically, the pancreas showed wide-spread necrosis, extensive infiltration of polymorphonuclear leukocytes, and nuclear inclusions in pancreatic acinar cells. Large numbers of adenovirus particles were demonstrable in the acinar cells by electron microscopy. This is the second case of adenovirus-associated pancreatitis we have found in rhesus monkeys and the first case in which we attempted to isolate the virus. Our data indicate that adenovirus pancreatitis is a distinct entity in rhesus monkeys and may occur with greater frequency than is generally believed.


Bile duct adenomas as liver nodules.

Bile duct adenomas as liver nodules. Bile duct adenomas can be difficult to differentiate at laparotomy from small metastatic tumor nodules in the liver. This can present a problem to the pathologist and surgeon relative to the advisability of attempted curative vs pallative surgery. We report two such cases. Review of our autopsy materials shows the incidence of bile duct adenoma to be higher than usually reported. Even though they all present as grayish-white, firm nodules, usually frozen section will reveal the true nature of the lesion. Occasionally, however, the lesions with active bile duct proliferation can cause problems in diagnosis, even after such examination. Accurate diagnosis by recognition of the entity and its histological characteristics may be very important in the surgical treatment of patients.


Mastectomy as an adjunct to combination chemotherapy. Three patients with metastatic breast carcinoma who had untreated locally advanced primary tumors were treated initially with combination chemotherapy followed by hygienic mastectomy. There was marked regression of the primary tumor in each case after chemotherapy, allowing for a technically simpler mastectomy without skin grafts. There were no serious postoperative complications, or delay in the resumption of systemic chemotherapy in any of them. The postoperative chemotherapy produced complete disappearance of the distant metastases and the patients remain clinically free of disease without local recurrence for 21, 10, and 7 months, respectively. One of these patients had inflammatory carcinoma and did well with this combined approach. These findings suggest a rationale for such an approach in patients with inflammatory carcinoma and may be applicable to patients with stage III breast cancer in whom the primary tumors are locally advanced and technically difficult to resect.


Amebic peritonitis following rupture of an amebic liver abscess. Successful treatment of two patients. We present two successfully treated cases of amebic peritonitis. Acute peritonitis secondary to intra-abdominal rupture of an amebic liver abscess is an infrequent but serious complication of invasive amebiasis. Its diagnosis should be considered in anyone with a suspected liver abscess, jaundice, or diarrhea in whom peritonitis develops. This diagnosis should be further suggested in the United States if the patient is a male and is of Mexican origin in areas where this racial group constitutes the majority of cases of amebic disease. Use of radioisotope liver scans and the demonstration of serum precipitins to Endamoeba histolytica may provide rapid evidence of invasive disease, although surgical intervention is often necessary to make a specific diagnosis. Emetine hydrochloride alone or followed by metronidazole combined with surgical drainage is the current treatment for amebic peritonitis.


Pseudolymphoma in renal allograft recipients. Five renal transplant recipients exhibited giant systemic lymphadenopathy shortly after transplantation. Biopsy specimens did not show Hodgkin's lymphoma. Immunosuppression was continued in all patients. In contrast to the rapidly fatal course of malignant lymphoma in transplant recipients, adenopathy in these five patients has uniformly resolved. Patients have been observed for 6 to 15 months with no evidence of residual disease. Interval biopsy specimens are not malignant. Each patient received antithymocyte globulin from a single lot for 10 to 21 days after transplantation. During administration, T cell lymphocytes were suppressed to 5% of control values. When lymphadenopathy occurred, T cell values rebounded to 371% of control values. Toxoplasmosis titers as well as viral cultures of lymph node biopsy specimens were negative. These data indicate a benign course of this histologically malignant disease and suggest a lymphoblastic rebound phenomenon to antithymocyte globulin.


Malignant histiocytomas.

Malignant histiocytomas. During the past ten years, the histiogenesis of malignant histiocytomas and a group of related benign and malignant lesions have been the source of speculation. Although of heterogeneous histological appearance, it is believed that there is a common cell of origin for these neoplasms--the histiocyte. From 1966 to 1974, 16 patients were encountered who had neoplasms that fell into the general group of malignant histiocytomas. These tumors were variously located in the extremities, head, chest wall, retroperitoneum, lung, spermatic cord, and lower abdomen. Surgical treatment included radical amputations, wide local excision, pulmonary lobectomy, and nephrectomy. Cobalt therapy and chemotherapy with vincristine sulfate, cyclophosphamide, doxorubicin hydrochloride, and chlorambucil were also used. Ten of 16 patients are alive after treatment, two are alive with metastatic disease, but four have died of malignant disease.


DNA repair processes in germ cells demonstrated in ejaculated sperms of rabbits treated with methyl methane sulfonate. Male rabbits were treated with a single i.v. injection of 22.5 mg/kg methyl methane sulfonate (MMS). 0--24 h later [3H]-thymidine was injected in both testicles. Incorporation of the isotope in germ cell DNA was demonstrated in ejaculated sperms. In controls labeled sperms were demonstrated first on day 40--43. These cells were in the preleptotene spermatocyte phase at the time of [3H]-thmidine injection. In rabbits treated with MMS significant radioactivity occurred in sperms collected from day 19 onwards. These cells were in late spermatocyte and early spermatid phase of maturation when [3H]-thymidine was injected. Incorporation of thymidine in these cell populations is interpreted as an expression of unscheduled DNA synthesis, a repair process initiated after chemical damage of germ cell DNA by MMS. The usefulness of the rabbit test system within the framework of conventional mutagenicity screening tests is discussed.


The effect of ascorbic acid on infection chick-embryo ciliated tracheal organ cultures by coronavirus. Chick embryo tracheal organ cultures showed increased resistance to infection by a coronavirus after exposure to ascorbate, while chick respiratory epithelium and allantois-on-shell preparations showed no increase in resistance to infection by an influenza virus or a paramyxovirus.


Size and secondary structure of avian myeloblastosis virus associated ribosomal RNA: comparison with cellular and precursor ribosomal RNA. Ribosomal RNA isolated from ribosomes present inside avian myeloblastosis virus (AMV) was characterized by electron microscopy using the formamide-urea spreading technique. The molecular weight and the secondary structures were compared with those of r-RNA and precursor r-NA isolated from host cells, the leukemic myeloblasts. The molecular weight of viral r-RNA (1.62 +/- 0.18 X 10(6) and 0.69 +/- 0.10 X 10(6)) and the molecular weight of cellular r-RNA (1.63 +/- 0.18 X 10(6) and 0.67 +/- 0.09 X 10(6)), the latter obtained from avian myeloblasts, were found to be identical and comparable with the molecular weight of chicken liver r-RNA. Likewise, the secondary structures of viral r-RNA were identical to those of cellular r-RNA. The postulated possible precursor character of viral r-RNA was excluded, since the molecules of viral r-RNA do not show any similarity to those of precursor r-RNA. Previously observed differences in behavior of viral and cellular (myeloblastic) r-RNA in sedimentation and electrophoretic mobility are discussed.


Effect of serum on the antiviral and anticellular activities of mouse interferon. Fetal bovine serum markedly decreased the ability of mouse L-929 interferon preparations to inhibit the formation of L-929 clones, but did not affect their ability to inhibit vesicular stomatitis virus (VSV) plaque formation in these cells. This dissociation of effects by interferon preparations indicates that: 1. the mechanism of action of interferon for its anticlonal antiviral activities is different; or 2. the molecule responsible for the anticlonal activity is a separate growth inhibitory factor.


Induction capacity and influence of dThdMP on thymidine kinase activity of type 1 and 2 strains of herpes simplex virus. The thymidine kinase inducing ability of 104 strains of herpes simplex virus was studied comparatively. A pronounced relationship was established between induction of the enzyme and the serotype of the strains. As a rule, the strains of serotype 2 are weaker inducer of dThd- and dCyd-kinase activity than serotype 1 strains. A certain parallelism exists between induction of both enzymes, however the activity of the thymidine kinase increases after infection with herpes simplex virus 4--5 times more than that of the dCyd-kinase. Adaptation of the strains to cell cultures only slightly modifies the inducing ability of the herpes simplex virus strains. The thymidine kinase activity induced by HSV-1 and HSV-2 differ from each other and are different from the cell enzyme with respect to their thermal stability at 40 degrees C. These differences are expressed more clearly in the presence of 480 muM dThdMP during inactivation. dThdMP stabilizes the type 1 but not the type 2 enzyme.


Neurovirulence in cynomolgus monkeys of enterovirus 71 isolated from a patient with hand, foot and mouth disease. Six cynomolgus monkeys were inoculated subcutaneously with enteroviurs 71 (E71), isolated from the stools of a patient with hand, foot and mouth disease (HFMD). Clinical symptoms were observed in three of the six monkeys. One monkey showed complete paralysis of the lower extremities and two animals showed weakness in the hind limbs 4 to 7 days after inoculation. Lesions were found in the central nervous system (CNS) of all monkeys. Mild to moderate vascular lesions, perivascular cuffings, degeneration and disappearance of the neurons and meningial lymphocytic infiltration were observed in the grey and/or white matter of the spinal cord, medulla oblongata, cerebral cortex and brain stem. No virus was recovered from the CNS or liver of any of the six monkeys. However, serum neutralizing antibody titers had risen in monkeys inoculated with E71.


[Reverse plaque formation by hog cholera virus inducing interference with VSV (author's transl)]. Infection of PK15 cells with various strains of Hog Cholera (HCV, togaviridae) induces a transient refractory state to VSV. The reverse plaque procedure is convenient for HCV titration of virulent, "chronic" and attenuated strains.


Spinal intramedullary glioblastoma with intracranial seeding. Report of a case. Approximately one year after partial removal of an intramedullary glioblastoma at T12, a patient was admitted to Wadsworth Veterans Administration Hospital with signs and symptoms of an intracranial tumor. We describe the unusual occurrence of intracranial seeding from spinal glioblastoma and review this rare condition and the possible mechanism involved.


[Oedematous optic neuropathy from Perhexilline maleate associated with peripheral polyneuropathy (author's transl)]. The authors report on a polyradiculoneuropathy related to long-term treatment with Perhexiline maleate (a drug used in France since 1973 for coronary insufficiency) whose presenting sign is a bilateral aedematous optic neuropathy. The mechanism of this effect on the optic nerves and on papilloedema is discussed; it seems not to be caused by a direct toxic action on the optic fibres but by the existence of chronic intracranial hypertension.


Effects of rapidly rotating shifts on sleep patterns and sleep structure. Six young adult males were assigned to a rapidly rotating shift work schedule (2 d 6 am-4 pm, 2 d 4 pm-10 pm, and 2 d 10 pm-6 am). They lived in the laboratory and completed two rotations. Their nonshift activities were ad libitum including sleep times. The patterns and structure of sleep were analyzed. Shift times systematically affected sleep patterns. Within shifts, Ss slept as late as possible prior to the morning shift, went to bed shortly after the night shift, and slept approximately midway between shifts on the afternoon shift. In transitioning to a new shift, the new shift time tended to determine sleep time. Sleep was significantly longer in transitions between shifts. Sleep structure was not markedly changed.


Spindle cell lipoma (report of 2 cases and differential diagnosis). "Spindle Cell Limpoma" is a benign tumor consisting of immature mesenchymal spindle cells with occasional collagen fibers, myxoid areas and lipocytes scattered throughout in varying numbers. Only focally it shows the typical appearance of a lipoma. Therefore it may be confused with liposarcoma, fibrosing lipoma, furthermore with areas of fibrous histiocytoma or nodular fasciitis showing fat infiltration.


Vitamin D metabolism and expression in rats fed on low-calcium and low-phosphorus diets. 1. Cholecalciferol, radioactively labelled with both (14)C and (3)H, was administered weekly for 7 weeks to rats that had been depleted of vitamin D for 4 weeks before repletion with the radioactive vitamin. This permitted measurement of the steady-state effect on vitamin D metabolism of low-calcium and low-phosphorus regimens, as compared with a normal mineral intake. These dietary manoeuvres were carried out during the last 3 weeks of repletion. Cholecalciferol, 25-hydroxycholecalciferol and 1,25-dihydroxycholecalciferol were determined in plasma, intestine, kidney and bone. Ca(2+)-binding-protein content was measured in intestine and kidneys of comparable animals. 2. In rats on the low-calcium diets, 1,25-dihydroxycholecalciferol concentration was elevated in plasma, bone, kidney and intestine, and intestinal Ca(2+)-binding protein was increased to over twice the concentration found in the control animals. 3. The low-phosphorus regimens led to a decrease in plasma phosphate and 1,25-dihydroxycholecalciferol in all tissues studied, for the latter to the point where it was undetectable in plasma and bone. Intestinal and renal concentrations of Ca(2+)-binding protein were unchanged in the low-phosphate-intake group and decreased in the very-low-phosphate-intake group. 4. It is concluded that in the rat, unlike in the chick, hypophosphataemia is not associated with a stimulation of the production of 1,25-dihydroxycholecalciferol or its expression in the synthesis of Ca(2+)-binding protein. Therefore the plasma phosphate concentration does not appear to be directly involved in the regulation of the functional metabolism of vitamin D.


Effects of pent-4-enoate on cellular redox state, glycolysis and fatty acid oxidation in isolated perfused rat heart. The metabolic effects of pent-4-enoate were studied in beating and potassium-arrested perfused rat hearts. The addition of 0.8mm-pent-4-enoate to the fluid used to perfuse a potassium-arrested heart resulted in a 70% increase in the O(2) consumption and a 66% decrease in the glycolytic flux as measured in terms of the de-tritiation of [3-(3)H]glucose, although the proportion of the O(2) consumption attributable to glucose oxidation decreased from an initial 30% to 10%. The pent-4-enoate-induced increase in O(2) consumption was only 15% in the beating heart. In the potassium-arrested heart, pent-4-enoate stimulated palmitate oxidation by more than 100% when measured in terms of the production of (14)CO(2) from [1-(14)C]palmitate, but in the beating heart palmitate oxidation was inhibited. Perfusion of the heart with pent-4-enoate had no effect on the proportion of pyruvate dehydrogenase found in the active form, in spite of large changes in the CoASH and acetyl-CoA concentrations and changes in their concentration ratios. The effects of pent-4-enoate on the cellular redox state were dependent on the ATP consumption of the heart. In the beating heart, pent-4-enoate caused a rapid mitochondrial NAD(+) reduction that subsequently faded out, so that the final state was more oxidized than the initial state. The arrested heart, however, remained in a more reduced state than initially, even after the partial re-oxidation that followed the initial rapid NAD(+) reduction. The ability of pent-4-enoate to increase or decrease fatty acid oxidation can be explained on the basis of the differential effects of pent-4-enoate on the concentration of citric acid-cycle intermediates under conditions of high or low ATP consumption of the myocardial cell. The proportion of the fatty acids in the fuel consumed by the heart is probably primarily determined by the regulatory mechanisms of glycolysis. When pent-4-enoate causes an increase in the citric acid-cycle intermediates, feedback inhibition of glycolysis results in an increase in the oxidation of fatty acids.


Quantitative electron-paramagnetic-resonance measurements of the electron-transfer components of the photosystem-I reaction centre. E.p.r. spectrometry was used to investigate the quantitative relationships between the oxidized chlorophyll free-radical signal I and the reduced iron-sulphur centre-A signal generated on illuminating Photosystem-I particles at cryogenic temperatures. In Photosystem-I particles prepared by using the French press or Triton X-100, at pH8.0 in the presence and absence of ascorbate and at pH 10.0 in the presence of ascorbate, the size of the light-induced signal I and iron-sulphur centre-A signals, corresponded to equal numbers of unpaired electron spins in each component. At 77K the spin-lattice relaxation time, T1, of the free radical signal I in samples of Photosystem-I particles prepared with Triton X-100 in the absence of ascorbate was 0.68 times the T1 value in the presence of ascorbate. Such changes in relaxation time can account for the different quantitative conclusions incorrectly arrived at from measurements made at saturating microwave powers [Bearden & Malkin (1976) Biochem. Biophys. Acta 430, 538-547; Malkin & Bearden (1976) FEBS Lett. 69, 216-220]. In the presence of benzoquinone and ferricyanide the ratio of free radical to centre A was 2.96:1, and at 77K the T1 was 0.50 times the T1 for ascorbate-treated samples. Here free radicals from bulk chlorophyll are generated in addition to those from the reaction-centre chlorophyll.


Quantitative electron-paramagnetic-resonance measurements of the electron-transfer components of the photosystem-I reaction centre. The reaction-centre chlorophyll (P700), the primary electron acceptor X and bound iron-sulphur centre A. An e.p.r. spectrum of the reduced form of the electron-transport component (X), thought to be the primary electron acceptor of Photosystem I, was obtained. By using line-shape simulations of this component and the free-radical e.p.r. signal I of the oxidized reaction-centre chlorophyll (P700), it was possible to determine the ratio of the number of electron spins to which these signals correspond in Photosystem-I particles under a variety of conditions. On illumination at cryogenic temperatures of Photosystem-I preparations, in which both bound iron-sulphur centres A and B were reduced, the measured ratio of free radical to component X varied between 1.04 and 2.23, with an average value of 1.54 +/- 0.18 where a Gaussian line-shape is assumed for the component-X signal in the simulation. The error in this measurement is estimated to be up to 50%. In a similar way component X and centre A of the bound iron-sulphur protein were quantified, the ratio between these two components varying between 1.26 and 0.61 with an average value of 0.75 +/- 0.06. These results indicate that the quantitative relationship, in terms of net electron spins, between centre A, component X and P700 is of the order to be expected if component X is indeed the primary electron acceptor in Photosystem I and a component of the photosynthetic electron-transport chain.


The oxidative activities of membrane vesicles from Bacillus caldolyticus. Energy-dependence of succinate oxidation. 1. The properties of membrane vesicles from the extreme thermophile Bacillus caldolyticus were investigated. 2. Vesicles prepared by exposure of spheroplasts to ultrasound contained cytochromes a, b and c, and at 50 degrees C they rapidly oxidized NADH and ascorbate in the presence of tetramethyl-p-phenylenediamine. Succinate and l-malate were oxidized more slowly, and dl-lactate, l-alanine and glycerol 1-phosphate were not oxidized. 3. In the absence of proton-conducting uncouplers the oxidation of NADH was accompanied by a net translocation of H(+) into the vesicles. Hydrolysis of ATP by a dicyclohexylcarbodi-imide-sensitive adenosine triphosphatase was accompanied by a similarly directed net translocation of H(+). 4. Uncouplers (carbonyl cyanide p-trifluoromethoxyphenylhydrazone or valinomycin plus NH(4) (+)) prevented net H(+) translocation but stimulated ATP hydrolysis, NADH oxidation and ascorbate oxidation. The last result suggested an energy-conserving site in the respiratory chain between cytochrome c and oxygen. 5. Under anaerobic conditions the reduction of cytochrome b by ascorbate (with tetramethyl-p-phenylenediamine) was stimulated by ATP hydrolysis, indicating an energy-conserving site between cytochrome b and cytochrome c. However, no reduction of NAD(+) supported by oxidation of succinate, malate or ascorbate occurred, neither did it with these substrates in the presence of ATP under anaerobic conditions, suggesting that there was no energy-conserving site between NADH and cytochrome b. 6. Succinate oxidation, in contrast with that of NADH and ascorbate, was strongly inhibited by uncouplers and stimulated by ATP hydrolysis. These effects were not observed when phenazine methosulphate, which transfers electrons from succinate dehydrogenase directly to oxygen, was present. It was concluded that in these vesicles the oxidation of succinate was energy-dependent and that the reoxidation of reduced succinate dehydrogenase was dependent on the outward movement of H(+) by the protonmotive force. 7. In support of the foregoing conclusion it was shown that the reduction of fumarate by NADH was an energy-conserving process. 8. If the activities of vesicles accurately represent those of the intact organism it appears that in B. caldolyticus the reduction of fumarate to succinate at the expense of reducing equivalents from NADH is energetically favoured over succinate oxidation even under aerobic conditions. This may be related to the need for an ample supply of succinate for haem synthesis in order to provide cytochromes for the organism.


Hyporesponsiveness of lymphocytes to virus antigens in rheumatoid arthritis. The immune response of peripheral blood lymphocytes to measles, rubella, parainfluenza types 1, 2, and 3 RNA virus antigens and to varicella-zoster and herpes virus type 1 DNA virus antigens was evaluated in 14 patients with rheumatoid arthritis (RA) and 14 matched controls by assessing 3H-thymidine incorporation. The results demonstrated hyporesponsivenss of lymphocytes from RA patients to virus antigens and phytohemagglutinin (PHA), which did not appear to be nonspecific because of a lack of correlation between response to virus antigens and response to mitogens. The patterns of decreased responsiveness was suggestive of a relative restriction of hyporesponsiveness to RNA virus antigens.


Immunopathologic studies of rheumatoid arthritis. I. Absence of complement-dependent cytotoxicity of rheumatoid sera for rheumatoid synovial cell cultures. A sensitive complement-dependent chromium release cytotoxicity assay was used to determine whether sera from rheumatoid arthritis (RA) patients contain antibody specific for an antigen on rheumatoid synovial cell cultures. Two hundred eight RA sera-RA synovial culture combinations were studied employing 21 sera and 16 synovial membranes; control combinations were derived from 5 normal sera and 10 degenerative joint disease synovial membranes. Anticomplementary activity of some rheumatoid sera was overcome using an increased complement concentration. The percent cytotoxicity of RA serum-RA culture combinations, both homologous and autologous, was not significantly greater than that of RA serum-control culture combinations. No correlation between duration of disease or duration of cell culture and percent cytotoxicity was found. Thus a unique antigen on cultured rheumatoid synovial cells was not recognized by rheumatoid serum antibody by use of this cytotoxicity assay.


[Enhanced bioavailability of digoxin from silica matrix formulations (author's transl)]. The bioavailability of digoxin from 3 silica matrix formulations was assessed in single-dose crossover studies in 12 healthy human volunteers: digoxin/silica matrix tablets (I, Digacin), digoxin/silica matrix in capsule form (II) and digoxin/silica matrix dragées, protected against gastrict juice by film coating (III). Urinary glycoside excretion for 6 days after 0.5 mg doses were measured by radioimmunoasay. Referring to an intravenous injection the bioavailability of digoxin from Digacin tablets is 82%, from the encapsulated matrix 69%, and from the dragées 54%. In comparison with corresponding results from other investigators Digacin tablets havet the same high bioavailability as digoxin solutions. In vitro liberations of digoxin from the silica matrix formulations (94% in 90 s) is significantly better than from conventional tablets produces from a digoxin-lactose trituration (61% in 90 s).


Tobramycin and neuromuscular transmission in the rat isolated phrenic nerve-diaphragm preparation. The effects of different concentrations of tobramycin, streptomycin, neomycin and gentamicin on the rat isolated phrenic nerve-diaphragm preparation are reported. Streptomycin, neomycin and gentamicin produced dose-dependent neuromuscular blockade. Tobramycin increased the muscle response at high concentrations (6.4 X 10(-4) - 2.6 X 10(-2) mol litre-1, but had no detectable effect when used in therapeutically recommended concentrations.


Factors affecting serum IgA antibody to Epstein-Barr viral capsid antigens in nasopharyngeal carcinoma. Irrespective of the ethnic origin of the patient, nasopharyngeal carcinoma (NPC), appears to stimulate the production of IgA antibodies against VCA. These antibodies are detected at high frequency and titres in sera from NPC patients but only rarely from control subjects. A majority of relapse-free survivors tested 1-12 years after radiotherapy (RT) sustain a detectable level of IgA anti-VCA. Serum titres of IgA anti-VCA remain relatively unchaged in individual NPC patients after RT, regardless of the disease evolution. These antibodies were detected in serum from one individual 9 months before NPC and the titre rose concomitantly with its clinical onset. Titres of IgA anti-VCA in multiple serum specimens from individual NPC patients, and in sera from different NPC patients, do not correlate with titres of IgG anti-VCA or with Serum IgA. It thus seems possible that the IgA anti-VCA in the sera of NPC patients might be largely derived near NPC. Apparently healthy individuals showing detectable IgA anti-VCA tend to aggregate in families of NPC patients. The distribution of siblings of these families who have the IgA anti-VCA reaction shows the binomial distribution expected for an autosomal recessive trait, implying the involvement of an autosomal recessive gene in the IgA anti-VCA response.


Cytotoxic effects of hexavalent and trivalent chromium on mammalian cells in vitro. The cytotoxic effects of hexavalent (k(2)Cr(2)O(7)) and trivalent (CrCl(3)) chromium compounds have been studied in cultured hamster fibroblasts (BHK line) and human epithelial-like cells (HEp line).K(2)Cr(2)O(7) stimulates the uptake of labelled thymidine into the soluble intracellular pool (the stimulation of nucleoside uptake represents a specific effect of Cr(6+)) while Cr(3+) always exerts an inhibitory action. DNA Synthesis is inhibited by treatment with both chromium compounds, but especially by K(2)Cr(2)O(7). Moreover, the effective CrCl(3) concentrations reduce the sensitivity of DNA and RNA to hydrolysis with perchloric acid. Treatments with k(2)Cr(2)O(7) in balanced salt solution, where Cr(6+) reduction is less marked, induce more pronounced cytotoxic effects than treatments in complete growth medium.HEp cells turned out to be more sensitive to K(2)Cr(2)O(7) than BHK fibroblasts: in the former line TdR uptake is less stimulated, DNA synthesis and cell survival are more affected. Survival of BHK cells to K(2)Cr(2)O(7) indicates a multi-hit mechanism of cell inactivation, the extrapolation number being about 10.On the basis of quantitative Cr determinations in the treatment solutions and in the treated cells, the cytotoxic effects of Cr are attributed to the action of Cr(6+) at the plasma membrane level on the mechanisms involved in nucleoside uptake, and to the interaction of Cr(3+) at the intracellular level with nucleophilic targets on the DNA molecule.


Binding of microorganisms to the macrophage plasma membrane; effects of enzymes and periodate. The nature of the binding of C. parvum organisms to the surface of glass-adherent mouse peritoneal exudate cells in vitro was studied using pretreatment of the cells with various enzymes and periodate. Trypsin, pronase, beta-galactosidase, phospholipases A, C and D and periodate all caused a decrease in binding to 40-60% of untreated control. Neuraminidase led to a 30% increase in binding. The binding ability returned to normal after 1 h at 37 degrees in culture medium following exposure to all the enzymes apart from pronase, which apparently could not be removed effectively by washing. The presence of EDTA in the medium inhibited recovery from treatment with trypsin and beta-galactosidase; return to normal after exposure to phospholipases A, C and D was slightly affected, whereas recovery from treatment with neuraminidase was unaffected. Cells that had been exposed to periodate did not regain normal binding ability after 1 h in tissue culture medium but the effect could be reversed chemically by treatment with borohydride. The role of different plasma membrane components in non-specific cellular recognition is discussed.


The relationship between cyclic adenosine 3', 5' - monophosphate and biochemical events in rat skin after the induction of epidermal hyperplasia using hexadecane. Sequential changes in skin metabolism have been studied in a model system of epidermal hyperplasia and hyperkeratinization induced by the application of n-hexadecane to shaved rat skin. The epidermal accumulation of glycogen typical of the hyperplastic response has been correlated with an increase in glycogenesis and a decrease in glycogenolysis. DNA synthesis was increased by 6 h after the start of hexadecane treatment and reached a maximum after one day. The concentration of skin cyclic AMP fell immediately after hexadecane application and subsequently rose to give a prolonged increase. Use of the combined topical application of hexadecane and the anti-inflammatory drugs triamcinolone acetonide, hydrocortisone and indomethacin showed that the hexadecane-induced changes in DNA synthesis and glycogen metabolism were linked to the initial fall in cyclic AMP concentration. The significance of the biphasic change in cyclic AMP levels is discussed as a possible system of control for the development and maintenance of hyperplasia.


Distribution of enzyme of nucleic acid metabolism in cow snout epidermis. Certain enzymes participating in the degradation of DNA have been measured in cow snout epidermis, liver, kidney and intestinal epithelium. High activities were found in epidermis compared to other tissues. The enzyme profile of the epidermis was investigated in relation to depth. Those enzymes directly concerned with DNA degradation occurred mainly in the upper (keratinizing) layer.


Some features of bone marrow macrophages in patients with homozygous beta-thalassaemia. The bone marrow macrophages of patients with homozygous beta-thalassaemia were frequently situated adjacent to collagen fibres and sometimes formed intrasinusoidal cytoplasmic protrusions. They also appeared to phagocytose processes of erythroblast cytoplasm (at times containing precipitated alpha-chains) which projected into them from neighbouring erythroblasts. The cytoplasm of the macrophages included large numbers of heavily-iron-loaded secondary lysosomes of various sizes and shapes in addition to phagocytosed erythroblasts, erythrocytes and extruded erythroblast nuclei. Numerous ferritin molecules were found in the cytoplasmic matrix but there were hardly any in the mitochondria, endoplasmic reticulum or golgi saccules. A small number of ferritin molecules were present within the nucleus. Electron microscope autoradiographs of marrow fragments which had been incubated with [3H]leucine for 1 h revealed the presence of newly-synthesized protein molecules in all types of secondary lysosomes. Light microscope autoradiographs showed the [3H]thymidine labelling index of the bone marrow macrophages was less than 1% and suggested that only a very small proportion of these cells were actively preparing for division.


Impairment of platelet thromboxane A2 generation and of the platelet release reaction in two patients with congenital deficiency of platelet cyclo-oxygenase. Two cases of thrombocytopathia with congenital deficiency of platelet cyclo-oxygenase were investigated. The platelet release reaction was impaired. There was a marked decrease of aggregation with collagen and with adrenalin and a total absence of aggregation with sodium arachidonate. The platelet response to labile aggregation stimulating substance (LASS, mostly thromboxane A2) was normal. There was no biosynthesis of prostaglandin cyclic endoperoxides or of thromboxane A2 from arachidonic acid. Basal levels of platelet PGE1 were lowered although plasma levels were normal. Thrombin decreased the cyclic AMP content of patients' platelets and also that of control platelets pretreated with aspirin. The patients platelets showed no ultrastructural difference when compared with control platelets, except for a slight decrease of granule volume, but, in contrast to control platelets, thrombin (0.02 U/ml) did not provoke contraction of the patients' platelets.


Intracellular lambda light chain inclusions in chronic lymphocytic leukaemia. A case of chronic lymphocytic leukaemia (CLL) with intracytoplasmic crystalline inclusions in peripheral blood lymphocytes is presented. The inclusions were identified as crystals of lambda (lambda) light chain and were present in bone marrow derived (B-cells) lymphocytes. The occurrence of light chain crystals in CLL may reflect a more primitive defect in immunoglobulin (Ig) synthesis, intracellular transport or secretion than has previously been reported.


The orientation of a heme of cytochrome c oxidase in submitochondrial particles. The electron paramagnetic resonance of the low spin signal from oxidized cytochrome c oxidase has been studied in oreinted multilayers of submitochondrial and electron transport particles. Measurements of the angular variation of the EPR spectra with the multilayer plane orientation allow the determination of the heme orientation in the multilayer. The heme normal lies in the membrane plane and the y-axis of the heme makes an angle of 30 degrees with the membrane normal. Analysis of the line shape reveals the presence of mosaic spread in the multilayer almost half of which is attributable to deviations of protein orientation within the membrane.


Theory of proton hyperfine interactions in bacterial photosynthetic systems. Bacteriopheophytin cation and anion. The electronic structures of the cation and anion of bacteriopheophytin alpha monomer are investigated by the self-consistent charge extended Hückel procedure including both pi and sigma electrons in the molecule. The calculated electron distributions are tested by comparison of the predicted hyperfine fields at proton sites with experimental data in both the ions and most important, by their ability to explain the observed trend in the hyperfine fields in going from the cation bacteriopheophytin+ alpha to the anion, a trend that is similar in many respects to the corresponding observed trend for the bacteriochlorophyll alpha cation and anion. Good agreement is obtained with experiment both for the absolute values of the observed proton hyperfine fields in both bacteriopheophytin a cation and anion as well as the ratio of the corresponding fields for the two systems. In particular, our calculated electron distributions in the two molecules lead, for the cation, to substantially different proton hyperfine fields for the two methyl groups attached to rings I and III, while for the anion, the corresponding fields are much closer to each other, a trend in good agreement with recent data. Also explained are the features of larger methine hyperfine constants in the anion as compared to the cation and the reverse trend for the protons in rings II and IV. Other features of the calculated electron distributions in the cation and anion are discussed and compared with each other. Possible additional measurements in the two systems that could provide further tests of the theoretically obtained electron distribution will be pointed out.


Resonance Raman spectra of cytochrome oxidase. Evidence for photoreduction by laser photons in resonance with the Soret band. Resonance Raman spectra of cytochrome oxidase solubilized in Tween 20 and sodium cholate, and excited at 413.1 nm have been recorded. Differences in the resonance Raman spectra of the two preparations are minimal indicating that the local environment of the hemes is similar in the two preparations. As in the work of Salmeen, et al. (1973) (Biochem. Biophys. Res. Commun. 52, 1100) the strongest band appears at 1358 cm-1. Some of the other bands differ slightly in their band shapes and frequencies when compared to their spectra; these differences can be accounted for by differences in resonance enhancement of the various bands wnen exciting at 441.6 and 413.1 nm. A study of the region from 1350 to 1380 cm-1 as a function of laser intensity (10--130 mW on sample) indicate that the doublet reported by Salmeen, et al. at 1358 and 1372 cm-1 is a result of photoreduction of the preparations. In samples to which potassium ferricyanide had been added, broad luminescence bands appear at 476 and 641 nm from which it is inferred that catalytic amounts of flavin in the preparations are photoreduced providing reducing equivalents to cytochrome oxidase.


Molecular motion and order in oriented lipid multibilayer membranes evaluated by simulations of spin label ESR spectra. Effects of temperature, cholesterol and magnetic field. A simulation method to interpret electron spin resonance (ESR) of spin labelled amphiphilic molecules in oriented phosphatidylcholine multibilayers in terms of a restricted motional model is presented. Order and motion of the cholestane spin label (3-spiro-doxyl-5alpha-cholestane) incorporated into egg yolk phosphatidylcholine, dipalmitoylphosphatidylcholine and dimyristoylphosphatidylcholine, pure and in mixture with cholesterol, were studied at various temperatures. With egg yolk phosphatidylcholine identical sets of motional parameters were obtained from simulations of ESR spectra obtained at three microwave frequencies (X-, K- and Q-band). With dipalmitoylphosphatidylcholine and dimyristoylphosphatidylcholine analyses of the spectra show that phase transitions occur in samples containing up to 30 mol % cholesterol. The activation energy for the motion of the spin label is about three times larger above than below the phase transition, indicating a more collective motion in the lipid crystalline state than in the gel state. In the liquid crystalline state the activation energy is larger in the pure phosphatidylcholines than with cholesterol added. Additions of cholesterol to egg phosphatidylcholine induces a higher molecular order but does not appreciably affect correlation times. This is in contrast to dipalmitoylphosphatidylcholine where both order and correlation times are affected by the presence of cholesterol. The activation energies follow the same order as the transition temperatures: dipalmitoylphosphatidylcholine greater than dimyristoylphosphatidylcholine greater than egg yokd phosphatidylcholine, suggesting a similar order of the cooperativity of the motion of the lipid molecules. Magnetic field-induced effects on egg phosphatidylcholine multibilayers were found at Q-band measurements above 40 degrees C. The cholestane spin label mimics order and motion of cholesterol molecule incorporated into the lipid bilayers. This reflects order and motion of the portions of the lipid molecules on the same depth of the bilayer as the rigid steroid portions of the intercalated molecules.


Chemically-induced cation permeability in red cell membrane vesicles. The sidedness of the response and the proteins involved. Cation fluxes were measured in right-side-out and inside-out vesicles obtained from human red cells. Rubidium, which is spontaneously released at very slow rates, can be rapidly released from both types of vesicle by addition  of valinomycin. P-Chloromercuriphenyl sulfonic acid (PCMBS) also increases the cation permeability of the vesicles with reversal to normal after addition of dithiothreitol. The effect of PCMBS is considerably larger and appears faster in the inside-out vesicles as compared to the right-side-out vesicles, the difference being greater at low temperatures. These data indicate that the SH groups responsible for the changes in cation permeability are more accessible from the inside face of the membrane. The response to PCMBS was not diminished after selective removal of extrinsic proteins by alkaline extraction, and/or after the membranes were exposed to proteolytic enzymes. The major polypeptide component remaining in vesicles after both treatments was a 17 000-dalton transmembrane fragment derived from band 3 which might, therefore, be responsible for the permeability response. Addition of Ca2+ to either right-side-out or inside-out vesicles, in the presence or absence of ionophore A23187, was without effect on monovalent cation permeability, indicating that the mechanism of Ca2+-induced K+ permeation was lost or inactivated during the preparation of the vesicles.


Plasma membrane protein kinase activity in normal and Rous sarcoma virus-transformed chick embryo fibroblasts. A preliminary study has been carried out to investigate the effect of Rous sarcoma virus transformation on plasma membrane protein kinase activity in chick embryo fibroblasts. Enzyme activity was measured using an in vitro phosphorylation method employing [gamma-32P]ATP with isolated plasma membranes serving as the source of both protein kinase and protein substrate. In general, the enzymatic properties observed were similar to those of other known protein kinases. However, for maximal activity a marked dependence on high Mg2+ concentrations was noted. Evidence was obtained which showed that cyclic nucleotide-dependent protein kinases were present in membranes from normal cells, but none could be measured in preparations from transformed cells. In addition, transformation appeared to result in a slight increase in basal plasma membrane protein kinase activity.


A calcium-activated polyphosphoinositide phosphodiesterase in the plasma membrane of human and rabbit erythrocytes. Haemoglobin-free human erythrocyte ghosts that were prepared in the presence of EDTA and were then exposed to Ca2+ showed a substantial loss of phosphatidylinositol phosphate and phosphatidylinositol diphosphate, measured either chemically or by loss of 32P from the lipids of prelabelled membranes. At the same time there was, as reported previously (Allan, D. and Michell, R.H., (1976) Biochim. Biophys. Acta 455, 824--830), and approximately equivalent rise in the diacylglycerol content of the membranes. Analysis of the 32P-labelled water-soluble material released during this process showed that the major products were inositol diphosphate and inositol triphosphate. No change was seen in the phosphatidylinositol or phosphatidate content of the membranes, and there was no Ca2+-activated loss of 32P from the phosphatidate of prelabelled membranes: this suggests that Ca2+ did not activate phosphoinositide phosphomonoesterases or phosphatidate phosphomonoesterase in human erythrocyte membranes. It is concluded that human erythrocyte membranes contain at their cytoplasmic surface a Ca2+-activated phosphodiesterase that is active against both phosphatidylinositol phosphate and phosphatidylinositol diphosphate. Rabbit erythrocytes also contained this enzyme, but in these cells there was also evidence for the presence of a Ca2+-activated phosphatidate phosphomonoesterase.


Optimum interaction of sterol side chains with phosphatidylcholine. The specificity of the interaction between the cholesterol side chain and egg phosphatidylcholine was precisely defined by examining the effect of three new analogues of cholesterol with modified side chains on the ordering of two steroid spin labels in liposomes. The complete side chain of cholesterol was shown to be required for maximum ordering. Sterols with side chains shorter or longer than cholesterol caused significantly less ordering.


Specific release of plasma membrane enzymes by a phosphatidylinositol-specific phospholipase C. The release of plasma membrane ecto-enzymes by a phosphatidylinositol-specific phospholipase C from Staphylococcus aureus was investigated. There was no effect on L-leucyl-beta-naphthylamidase, alkaline phosphodeisterase I and Ca2+- or MG2+-ATPase, but substantial proportions of the alkaline phosphatase and 5-nucleotidase were released. There was no simultaneous release of phospholipid and the solubilized enzymes were not exluded from Sepharose 6-B. It was therefore concluded that release was not a secondary consequence of membrane vesiculation but occurred as a result of the disruption of specific interactions involving the phosphatidylinositol molecule.


The measurement of phosphatidate phosphohydrolase in human amniotic fluid. Phosphatidate phosphohydrolase (EC 3.1.3.4) activity can be found in late gestational human amniotic fluid and is thought to originate in type II alveolar cells of the fetal lungs where it plays an important role in lung surfactant synthesis. In the present study, phosphatidate phosphohydrolase activity was detected and characterized in a 105 000 X g pellet of amniotic fluid using either [32P]phosphatidate or a water-soluble analog, 1-O-hexadecyl-rac-[2-(3)H]glycerol 3-phosphate as substrate. With either substrate, enzyme activity was optimal at pH 6.0. The soluble analog was hydrolyzed with a Km value of 163 micrometer and a V of 30 nmole/min per mg of protein, and offered several advantages over phosphatidate as a substrate for assaying phosphatidate phosphohydrolase in amniotic fluid. Using the synthetic analog, phosphatidate phosphohydrolase activity was measured in the 700 X g supernatant fraction of 30 human amniocentesis samples and compared with another index of fetal lung maturity, the phosphatidylcholine/sphingomyelin ratio. The results suggest that the new phosphohydrolase assay may be clinically useful in the assessment of fetal lung development.


The effect of corticotropin on phospholipid metabolism in isolated adrenocortical cells. Trypsin-dispersed cat adrenocortical cells were incubated at 37 degrees C in modified Eagle's medium containing [14C]arachidonic acid of sodium [14C]-acetate and then in non-radioactive medium. Radioactive incorporation was obtained in all phospholipids, with the greatest amount of radioactivity in phosphatidylcholine, followed by phosphatidylethanolamine, phosphatidyl-serine, and phosphatidylinositol. Concentrations of individual phospholipids generally paralleled the relative amounts of corresponding radiolabeled phospholipids, although the percentage of phosphatidylinositol was considerably lower than its radioactive counterpart, resulting in a high specific activity of this particular phospholipid. Although a potently steroidogenic concentration of corticotropin failed to enhance release of label from any particular phospholipid, analysis of specific activity showed that corticotropin stimulation was accompanied by an increased turnover of phosphatidylinositol and phosphatidic acid. These studies demonstrate that isolated cortical cells have the capacity to synthesize phospholipids from radioactive precursors. The finding that the acute effects of corticotropin are associated with changes in specific phospholipids, including phosphatidylinositol and phosphatidic acid, conforms to the general pattern observed in other secretory systems.


Effects of ruthenium red, A23187 and D-600 on steroidogenesis in Y-1 cells. The effects of the calcium antagonists ruthenium red and D-600 and the cation ionophore A23187 on steroidogenesis were investigated. Steroidogenesis triggered by corticotrophin and cyclic AMP was inhibited by each of the agents. Incubation of Y-1 cells with an excess of ethyleneglycol-bis-(beta-amino-ethylether)-N,N'-tetraacetic acid (EGTA) abolished the steroidogenic response to corticotrophin while the response to cyclic AMP was unaffected. The ability of ruthenium red and D-600 (1 . 10(-5) M), and A23187 (6 . 10(-6 M) to inhibit a response which does not require the presence of extracellular calcium (cyclic AMP induced steroidogenesis) suggests that they are altering intracellular calcium. Neither of the calcium antagonists nor the cation ionophore inhibited the steroidogenic response to exogenous pregnenolone, thereby suggesting that the cells were still viable. Only when A23187 was used in the presence of a 15-fold increase in extracellular calcium (4.8 mM) was the response to pregnenolone diminished. The data are interpreted as a further indication that, in intact cells, intracellular calcium plays a role in the steroidogenic pathway.


Regulation of cholesterol ester synthesis in cultured glial and neuronal cells. Relation to control of cholesterol synthesis. Regulation of cholesterol ester synthesis was studied in cultured C-6 glial and neuroblastoma cells. Particular emphasis was placed on the relation of this regulation to control of cholesterol synthesis. The effectors studied were low density lipoprotein (LDL) and desmosterol. Distinct differences in regulation were observed between the glial and neuronal cells. In the neuronal cells cholesterol ester synthesis (from [14C]oleate) was not affected by even high concentrations of LDL or desmosterol. In contrast, cholesterol ester synthesis was stimulated as much as 12-fold in the glial cells after just 5 h exposure to LDL or desmosterol. Cholesterol synthesis (from [14C]acetate) was inhibited in a simultaneous and quantitatively similar manner. Suitable experiments indicated that alterations in pool sizes of intermediates did not contribute to the genesis of the observed responses and suggested that LDL and desmosterol produced their effects by stimulating esterification of primarily endogenous cholesterol. The data may have major implications concerning the controlling metabolic events prior to and at the onset of myelination.


Suppression of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity and of incorporation of acetate into cholesterol in homozygous hypercholesterolemic fibroblasts by ferritin-low density lipoprotein conjugates. Conjugates of ferritin with low density lipoproteins (LDL) were prepared and separated by sucrose gradient centrifugation. These conjugates, at cholesterol concentration of 100--132 microgram/ml, caused a greater than 90% suppression of hydroxymethylglutaryl coenzyme A reductase activity and of acetate incorporation into cholesterol in cultured skin fibroblasts from a normal subject as well as from a subject with homozygous familial hypercholesterolemia. The half maximal inhibition concentration was approx. 10 microgram/ml cholesterol for LDL and ferritin . (LDL)2 and 5 microgram/ml for (ferritin)2 . LDL in both cell lines. In contrast, native low density lipoproteins have only a minimal inhibitory effect in homozygous cells. The ability of the conjugates to stimulate the incorporation of oleate into cholesteryl esters was also equal in the two cell lines, although the conjugates were only 10% as active as low density lipoproteins in the normal cells. LDL reduced the ferritin . (LDL)2-mediated suppression of hydroxymethylglutaryl-CoA reductase activity in homozygous cells while ferritin . (LDL)2 reduced the LDL-mediated stimulation of cholesteryl ester formation in normal cells.


Proteolysis of very low density lipoprotein in perfused lung. Perfusion of homologous 125I-labeled rat very low density lipoprotein through isolated rat lungs in the presence of heparin resulted in apoprotein proteolysis. At least the apoprotein C was degraded into two peptides smaller than 7500 daltons as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The lung uptake of radioactivity was small and due mainly to the presence of the larger of the two peptides. The lung protease was not active against an 125-I-labeled albumin substrate and was not released into the medium by heparin.


Prostaglandin biosynthesis and catabolism in the lamb ductus arteriosus, aorta and pulmonary artery. Homogenates of tissues from fetal and neonatal lamb ductus arteriosus, aorta and pulmonary artery have the capacity to convert arachidonic acid as well as the intermediate prostaglandin endoperoxide, prostaglandin H2, into three products: prostaglandins E2, F2alpha and a major product 6-ketoprostaglandin F1alpha. The three tissues also displayed prostaglandin 15-hydroxydehydrogenase and 13-reductase catabolic activities. The catabolishing system showed considerable substrate specificity: prostaglandin E1 was a good substrate whereas prostaglandins F1alpha and F2alpha were completely devoid of catabolism. The complete system was observed in immature as well as mature arterial vessels, in the fetus as well as the neonate (up to 7 days old). These experiments demonstrate the presence of several components of the prostaglandin system in these tissues and offer biochemical evidence for the implication of prostaglandins E2 and I2 in the maintenance of the ductus and neighboring vessels in a relaxed state in the fetus.


Induction of low density lipoprotein receptor synthesis by high density lipoprotein in cultures of human skin fibroblasts. Further studies have been made of the effects of high density lipoprotein (HDL) on the surface binding, internalization and degradation of 125I-labeled low density lipoprotein (125I-labeled LDL) by cultured normal human fibroblasts. In agreement with earlier studies, during short incubations HDL inhibited the surface binding of 125I-labeled LDL. In contrast, following prolonged incubations 125I-labeled LDL binding was consistently greater in the presence of HDL. The increment in 125I-labeled LDL binding induced by HDL was: (a) associated with a decrease in cell cholesterol content; (b) inhibited by the addition of cholesterol or cycloheximide to the incubation medium; and (c) accompanied by similar increments in 125I-labeled LDL internalization and degradation. It is concluded that HDL induces the synthesis of high affinity LDL receptors in human fibroblasts by promoting the efflux of cholesterol from the cells.


Enhanced degradation of trypsin-treated low density lipoprotein by fibroblasts from a patient with homozygous familial hypercholesterolemia. When 125I-labeled native low density lipoprotein was incubated with skin fibroblasts from a patient with homozygous familial hypercholesterolemia, the observed rate of degradation of the protein moiety was less than 5% the rate observed with normal fibroblasts, in agreement with previous studies. When the low density lipoprotein had been first treated with trypsin, with release of about 20% of the protein, its degradation by the patient's fibroblasts was markedly increased 8-20-fold. In contrast, the rate of degradation of the trypsin-treated lipoprotein by normal fibroblasts was, if anything, slightly reduced. In neither the normal cells nor the patient's cells was binding to the cell surface appreciably altered by trypsin treatment of the lipoprotein. Prior incubation with cholesterol and 7-ketocholesterol reduced binding of trypsin-treated low density lipoprotein to normal cells by 67% but did not affect its binding to the patient's cells. The results show that the structural modifications induced by trypsin do not interfere with binding of low density lipoprotein to its normal high affinity receptor nor its degradation by normal cells. However, the modified lipoprotein is much more readily internalized and degraded by cells from the patient with homozygous familial hypercholesterolemia.


The effects of dibutyryl cyclic adenosine 3':5'-monophosphate on concanavalin A-stimulated sterol and fatty acid synthesis in mouse spleen lymphocytes. Substantial increases in both 3beta-OH sterol and fatty acid synthesis were observed after concanavalin A addition to mouse spleen lymphocytes cultured in serum-free media. The rate of sterol synthesis increased linearly up to 60 h. The rate of fatty acid synthesis increased up to 20 h, reaching a plateau in synthetic activity which was the maintained. CO2 production from acetate was slightly stimulated by concanavalin A. In contrast to sterol and fatty acid synthesis, the rate of CO2 production in both mitogen-stimulated and resting cultures declined with time. Dibutyryl cyclic AMP had a strong inhibitory effect on concanavalin A-stimulated sterol and fatty acid synthesis from acetate, but only a slight effect on CO2 production. Delayed addition of dibutyryl cyclic AMP resulted in reduced inhibition. The data suggest a sequence of initiation for fatty acid and sterol synthesis prior to DNA synthesis and a possible regulatory role of cyclic AMP in this initiation. The results support the hypothesis that lymphocyte activation is sequential within the spleen cell population and is accompanied by fatty acid and sterol synthesis.


Solubilization and separation of delta5,3beta-hydroxysteroid dehydrogenase and 3-oxosteroid-delta4-delta5-isomerase from bovine adrenal cortex microsomes. A physical separation of delta5,3beta-hydroxysteroid dehydrogenase and 3-oxosteroid delta4-delta5-isomerase solubilized from bovine adrenocortical microsomes is described for the first time. The solubilization as well as the separation was carried out with a mixture of a detergent: a substituted betaine (Empigen BB/P) and sodium cholate. This latter detergent protects isomerase from complete inactivation by Empigen and is necessary for the recovery of a significant amount of soluble isomerase. Separation of dehydrogenase and isomerase was successfully accomplished by the use of a DEAE-Biogel A anion-exchanger. Dehydrogenase activity was eluted, while the isomerase was retained. Measurements of dehydrogenase activity with androst-5-en-3beta-ol-17-one, pregnen-3beta-ol-20-one and pregn-5-en-(3beta,17alpha)-diol-20-one and of isomerase activity with androst-5-en-(3,17)-dione and pregn-5-en-(3,20)-dione suggested that more than one isomerase and more than one dehydrogenase form were present.


Comparative study on glycolipid composition between two cell types of rat ascites hepatoma cells. A comparative study of the glycolipids was performed on two cell lines of rat ascites hepatomas, island-forming and free cell types, and several marked differences were found as follows: 1. Hexose content in glycolipids derived from AH 7974F (free cell type) was about 3.6-fold as much as that from AH 7974 (island-forming cell type) on the basis of dry cell weight. 2. The glycolipids in the cells of AH 7974 were tentatively identified as glucosylceramide, lactosylceramide, galactosylgalactosylglucosylceramide, globoside and hematoside (GM3) by both thin-layer and gas-liquid chromatography. 3. On the other hand, the glycolipids in AH 7974F cells were glucosylceramide, lactosylceramide and at least four unknown lipids. 4. Structure of one of these unknown lipids was shown to be asialo-GM2 by methylation and enzymatic degradation studies. Moreover, the presence of asialo-GM1 was suggested by immunoprecipitation test with anti-asialo-GM1 serum. 5. Glucosylceramide and lactosylceramide from AH 7974F cells were found to possess hydroxy fatty acids as major fatty acids components, which were rare in these glycolipids from AH 7974 cells.


A comparative spectroscopic study of two non-haem iron proteins lacking labile sulphide from Desulphovibrio gigas. The ultraviolet visible, and near infrared spectrum of a two-iron protein from Desulphovibrio gigas, a new type of non-haem iron protein lacking labile sulphide, is compared with that of D. gigas rubredoxin. The charge transfer band maxima of rubredoxin at 495 and 565 nm are less separated in the new protein implying a higher symmetry of the two iron centres. The existence of a spin-spin interaction between the two iron centres in the new protein is suggested by the magnetic susceptibility measurements of the oxidized and reduced states of both proteins, which gives a smaller value per iron centre for the new protein. The oxidized form of the two iron-protein has a complex EPR spectrum with signals at g values of 8.97, 7.72, 5.73, 4.94, and 1.84. An EPR titration gives a value of --35 +/- 15 mV for the two signals at g values of 7.72 and 5.73. Rubredoxin has the characteristic spectrum of rubredoxins with two signals at g values of 9.4 and 4.27.


Oxygenation and EPR spectral properties of Aplysia myoglobins containing cobaltous porphyrins. Cobalt myoglobins (Aplysia) have been reconstituted from apo-myoglobin (Aplysia) and proto-, meso-, and deutero-cobalt porphyrins. Each of them showed the 30--60 times lower oxygen affinity than those of the corresponding cobalt myoglobins (Sperm whale). Kinetic investigation of their oxygenation by the temperature-junp relaxation technique showed that the low oxygen affinity of cobalt myoglobin (Aplysia) is due to a large dissociation rate constant. the electron paramagnetic resonance (EPR) spectrum of oxy cobalt myoglobin (Aplysia) is affected by the replacement of H2O with D2O, suggesting a possible interaction between the bound oxygen and the neighboring hydrogen atom. A low temperature photodissociation study showed that the product of photolysis of oxy cobalt myoglobin (Aplysia) gives an EPR spectrum different from that of the deoxy-cobalt myoglobin (Aplysia) and from that of the photolysed form of oxy-cobalt myogloin (Sperm whale). These observations suggest that in oxy-cobalt myoglobin (Aplysia) the bound oxygen might interact with amino acid adjacent to it, but the interaction is weaker than that in oxy cobalt myoglobin (Sperm whale).


Assignment of the heme c resonances in the 360 MHz H NMR spectra of cytochrome c. In the 360 MHz 1H NMR spectra of horse heart ferrocytochrome c recorded after suitable digital resolution enhancement, the resonances of all the heme c protons with the exception of those of the propionic acid side chains were observed as well resolved lines. From spin decoupling and nuclear Overhauser effects in homonuclear double resonance experiments, all these resonances were assigned to their respective positions in heme c. With saturation transfer experiments in solutions of partially reduced cytochrome c, individual assignments were further obtained for the six heme c methyl resonances in ferricytochrome c. The present experiments add individual assignments to the earlier identifications of the heme c ring methyl and meso-proton resonances, and show that the earlier identifications of the thioether bridge methyl resonances must be revised. These data provide a basis for more detailed descriptions of the electronic structure of heme c and its possible relations with the pathway of the electron transfer in and out of the cytochrome c molecule. Furthermore, the pseudocontact shifts of the thioether bridge methyl resonances could be related to the electronic g-tensor measured by EPR in ferricytochrome c single crystals at low temperature. From this it will now be possible without chemical modification of the protein, to compare in detail the solution conformations near the heme c in reduced and oxidized cytochrome c and thus hopefully to obtain additional insights into the mechanism of the biological redox reaction of this protein.


Cleavage of native type III collagen in the collagenase susceptible region by thermolysin. Viscometric assays were used to demonstrate the activity of thermolysin (EC 3.4.24.4) on native type III collagen in solution. Analysis of the reaction products by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and electron microscopic visualisation of segment long spacing aggregates demonstrated localised cleavage of the collagen in the collagenase susceptible region.


An adenosine 3':5'-monophosphate-adenosine binding protein from mouse liver: some physicochemical properties. A number of physiochemical properties of the cyclic AMP-adenosine binding protein of mouse liver (Ueland, P.M. and Døskeland, S.O. (1977) J. Biol. Chem. 252, 677--686) have been studied. 1. The specific extinction coefficient, E1%280nm, was estimated to 13.0. 2. Amino acid and amide group analyses confirmed the acidic properties of the protein as determined by electrofocusing (pI = 5.7). Based on the estimated partial specific volume (v = 0.74 cm3/g) the minimum molecular weight of the native, tetrameric protein was recalculated to be 185 000 (s20,w = 8.8 . 10(-13) s and Stokes radius = 48 A). 3. No NH2-terminal amino acid was found by the dansyl method using [14C]-dansyl chloride, indicating that the NH2-terminal groups are blocked. 4. Amino acid analyses gave 6 half-cystine residues per subunit, and the same number of free sulfhydryl groups was found by titration of the denatured protein with 5,5'-dithiobis (2-nitrobenzoic acid). 5. The reactivity of the SH groups in the native protein with 5,5'-dithiobis (2-nitrobenzoic acid) revealed rapidly reacting (SHI), sluggishly reacting (SHII) and "masked" (SHIII) SH groups. ATP, adenosine, Mg2+ and KCl, factors known to affect the activation of cyclic AMP binding sites (Ueland, P.M. and Døskeland, S.O. (1978) Arch. Biochem. Biophys., in press) changed the reactivity of separate SH groups.


Removal of nonionic detergent from proteins fractionated by electrofocusing. The nonionic detergent Nonipol TD 12 (an alkyl polyoxyethylene alcohol) has been removed from solubilised proteins after their fractionation by electrofocusing. Following electrofocusing in nonionic detergent an anionic or cationic detergent was added to the focussing medium and the focusing was allowed to continue. The ionic detergent formed mixed micelles with the nonionic detergent. Thus charged, the mixed micelles migrated to either electrode, removing nonionic detergent from the fractionated proteins. Applying this technique to an adipose tissue preparation, detergent-inhibited activity of a lipolytic enzyme was restored and the binding of adenosine 3':5'-cyclic monophosphate (cyclic AMP) to a protein kinase was increased.


Multiple forms of cytosol and membrane-bound protein kinase activity in human erythrocytes. Both cytosol and membranes of human erythrocytes display protein kinase activity towards exogenous protein substrates such as casein, phosvitin and histones. The histone kinase activity, unlike casein kinase, of both cytosol and membranes is increased by cyclic AMP. The protein kinase forms removed from the membranes with 0.7 M NaCl, phosphorylate only serine residues of both casein and histones through a mechanism cyclic AMP-independent. The protein kinase activity located in the cytosol (hemolysate) is due also to enzyme forms phosphorylating both serine and threonine residues of casein, in addition to forms phosphorylating only serine residues of casein and histones. Also the cytosol kinase forms, once partially purified by Sepharose 6B filtration, appear to be cyclic AMP-independent.


Induction of cyclic AMP-binding proteins by dibutyryl cyclic AMP in mouse neuroblastoma cells. Mouse neuroblastoma cells grown in the presence of 1 mM N6,O2'-dibutyryl-cyclic AMP showed a 3-fold increase in cyclic AMP-binding proteins. The role of dibutyryl cyclic AMP in the introduction of cyclic AMP-binding proteins in these cells has been studied. Induced cyclic AMP-binding proteins were observed in the cytoplasm 15 h after dibutyryl cyclic AMP treatment. The increase in cyclic AMP-binding proteins required RNA and protein synthesis. It is suggested that the 15-h lag occurs at the post-transcriptional and/or translational level. Cyclic AMP-binding proteins are found in both soluble and particulate cell fractions. Dibutyryl cyclic AMP increased binding proteins in both fractions. The control and dibutyryl cyclic AMP-induced binding proteins showed similar affinity for cyclic AMP. The data indicate that dibutyryl cyclic AMP caused the following sequential events: a 12-fold increase in cyclic AMP levels; a 40% increase in phosphodiesterase activity; and a 300% (3-fold) increase in cyclic AMP-binding proteins. It is suggested that the differentiation of mouse neuroblastoma cells involves increased levels of cyclic AMP and cyclic AMP-binding proteins.


Synthesis and properties of a photoaffinity probe for the glucagon receptor in hepatocyte plasma membranes. The reaction of glucagon with 4-fluoro-3-nitrophenylazide has been shown to afford the photosensitive derivative, Nepsilon-4-azido-2-nitrophenyl-glucagon. The structure and properties of this derivative were established by amino acid analysis, absorption and fluorescence spectroscopy, deamination, Edman degradation and photolysis. This photoaffinity derivative of glucagon has been used to label specifically glucagon binding sites on hepatocyte plasma membranes.


beta-Adrenergic receptors in rat skeletal muscle. Effects of thyroidectomy. Thyroid hormones may participate in the regulation of beta-adrenergic receptors in skeletal muscle sarcolemmal membrane. Since skeletal muscles are not innervated by sympathetic nerve endings, the biochemical mechanism involved in the control of beta-adrenergic receptors by thyroid hormones appears to be mediated by thyroid-induced regulation of serum levels of catecholamines.


Inhibitory effects of glucocorticoids on collagen synthesis by mouse sponge granulomas and granuloma fibroblasts in culture. The basis for the glucocorticoid-mediated decrease in tissue collagen was studied in mouse granulomas and in primary granuloma fibroblast cultures. Injection of mice for 12 days with dexamethasone (0.35 mg/kg body weight) resulted in a 50--70% inhibition of collagen synthesis and accumulation in polyvinyl sponge-induced granulomas whereas total protein synthesis was inhibited by only about 25%. The decreased collagen content of the granuloma was accounted for by both a reduced fibroblast number and diminished synthesis per cell. Growth rates, total protein synthesis and collagen synthesis were the same in granuloma fibroblast cultures derived from control or steroid-treated mice. However, addition of 3.10(-7) M hydrocortisone to the culture medium caused a 30--50% inhibition of both collagen and non-collagen protein synthesis in firbroblasts from either source. These inhibitory effects were dose- and time-dependent with a lag time of 12--24 h. Prolyl hydroxylase activity was reduced both in sponge granulomas from glucocorticoid-treated mice and in hydrocortisone-treated fibroblast cultures. However, protein synthesis was inhibited to the same extent as the inhibition of prolyl hydroxylase activity and there was no effect on peptidyl prolyl hydroxylation. These results indicate that the glucocorticoid-induced reduction of collagen synthesis and accumulation observed in mouse granulomas and primary granuloma fibroblast cultures is not specific for this protein. Furthermore, glucocorticoid-induced inhibition of collagen synthesis cannot be attributed to underhydroxylation of collagen prolyl residues.


Endogenous substrates of protein kinase in rat liver cell sap under different dietary conditions. Liver cell sap from normally fed rats, rats fed with a high-carbohydrate diet and fasted rats was chromatographed on DEAE-cellulose (pH 7.0). The chromatogram from each diet group was analyzed for pyruvate kinase activity and endogenous substrates of cyclic AMP-stimulated protein kinase. The materials were pooled into five phosphorylatable fractions, in each of which phosphate incorporation at 0.1 mM and 1.0 mM [32P]ATP in the presence of cyclic AMP and protein kinase was determined. For characterization of the phosphorylatable components, thin-layer gel chromatography on Sephadex G-200 and polyacrylamide gel electrophoresis in detergent were used for determination of native and minimal molecular weights, respectively. Except for pyruvate kinase, eight components which incorporated at least 0.05 nmol of [32P]phosphate/g of liver were detected. The phosphorylation of four of them was stimulated by cyclic AMP. Their minimal molecular weights were 42000, 21000, 52000 and 49000. The component with a minimal molecular weight of 42000 seemed to have a native molecular weight of 160000. Both the 21000 and the 52000 component had a native molecular weight of about 110000-120000. The protein with a minimal molecular weight of 49000 could not be correlated with certainty to a native molecular weight. The proteins whose phosphorylation was not stimulated by cyclic AMP had minimal molecular weights of 54000, 39000, 34000 and 22000.


Cyclic AMP-mediated activation of hepatic glycogenolysis by fructose. Isolated livers from fed and fasted rats were perfused for 30 min with recirculating blood-buffer medium containing no added substrate and then switched to a flow-through perfusion using the same medium for an additional 5, 10 and 30 min. Continuous infusion of fructose for the final 5, 10 or 30 min resulted in activation of glycogen phosphorylase, an increase in the activity of protein kinase, elevated levels of tissue adenosine 3', 5'-monophosphate (cyclic AMP), and no consistent effect on glycogen synthase. Infusion of glucose under the same conditions resulted in activation of glycogen synthase, inactivation of glycogen phosphorylase, no change in protein kinase, and no consistent change in tissue cyclic AMP. These results demonstrate that while glucose promotes hepatic glycogen synthesis, fructose promotes activation of the enzymatic cascade responsible for glycogen breakdown.


Kinetics of cyclic AMP in toad bladder.

Kinetics of cyclic AMP in toad bladder. Tissue distributions of cyclic [3H]AMP and [14C]inulin in toad bladder were determined and their kinetics analyzed. We found that both the epithelial and the other cells of the toad bladder handle cyclic AMP similarly. Moreover, we found that the distribution of cyclic AMP did not differ from that of inulin, an extracellular marker. Kinetic analysis suggests that the rate coefficient of cyclic AMP metabolism is much larger than the exchange rate coefficient, which explains why distribution of both cyclic AMP and inulin are similar.


Electron spin resonance studies of the solution structure of vanadyl amino acid complexes and mixed ligand complexes of oxalate. Esr and electronic spectra of complexes of the general composition VO(AA)2 and VO(ox)(AA) have been characterized; AA = gly, his, cys, pro, val, met, asp amino acids. Spectra of the formulation VO(ox)(LL) (with LL = imidazole plus monodentate oxalate, histamine plus monodentate oxalate, histidine, cysteine, 4-imidazolepropionic acid, mercaptopropionic acid, ethylenediamine and ethanolamine) have been used to deduce a self-consistent assignment of AL, a ligand donor additivity constant contribution to the observed hyperfine splitting, Aiso. Values of AL are sensitive to inductive effects in the ligand structure. The solution structures and likely coordination geometries of VO(his)2, and VO(cys)22-- are discussed. The role of the imidazole moiety as a sigma donor and sulfhydryl sulfur as a pi acceptor is observed in VO(AA)2 and VO(ox)(AA) complexes.


Ontogeny of adenylate cyclase and phosphodiesterase activities in swine tissues. The ontogenic patterns of the basal- and fluoride-stimulated adenylate cyclase and the cAMP phosphodiesterase activites as well as the tissue levels of cAMP were assessed in swine adipose tissue, heart, skeletal muscle, and liver. The developmental patterns were complex. However, there was a general tendency in all tissues toward lower enzyme activities in the oldest animals (150 days) for both enzymes. In adipose tissue, the decreased enzyme levels could partially be attributed to an increase in adipocyte size.


P-chloromercuribenzene sulfonate blocks and reverses the effect of amiloride on sodium transport across rabbit colon in vitro. The addition of 10(-3) M p-chloromercuribenzene sulfonate (PCMBS) to the solution bathing the mucosal surface of rabbit colon has no effect on the rate of active Na transport but blocks or reverses the inhibitory action of amiloride. The tissue must be exposed to PCMBS for 20-30 min for a complete blocking effect, and removal of PCMBS from the mucosal solution after this period of exposure does not restore the sensitivity of the tissue to amiloride. The slow time-courses of the blocking and reversal effects suggest that PCMBS does not irreversibly interact with groups directly involved in the binding of amiloride.


Surface proteolytic activity in cell interactions: activation of human peripheral lymphocytes in contact with mitogen-treated glass-adherent leukocytes. The transformation of lymphocytes by mitogens or by periodate treatment requires the presence of glass-adherent leukocytes. We have demonstrated that contact with periodate-treated adherent cells transforms human peripheral T lymphocytes. Using a new method to estimate proteolytic activity released by cells or available at cell surfaces, we found marked stimulation of cell surface-associated enzyme activity on glass-adherent cells treated by NaIO4 or by Con A. The expression of this enzymatic stimulation required contact with substrate, just as the transformation of untreated T cells required contact with treated adherent cells. Therefore, we propose that cell-cell interaction leads to an efficient mode of stimulation, which may be mediated by or initiated by surface-associated proteases of adherent cells.


Specific alpha-adrenergic receptor desensitization in vascular smooth muscle. Specific desensitization of vascular tissue to norepinephrine was studied using an open-ring-type strip preparation of rabbit aorta. Aortae were taken from untreated, reserpine-pretreated (0.1 mg/kg/day; 3, 5, or 7 days) and 6-hydroxydropamine-pretreated (50 mg/kg; 3 or 7 days before sacrifice) rabbits. Aortic strips from the pretreated rabbits were supersensitive to norepinephrine. Subsequent to obtaining the first dose-response curve for norepinephrine, aortic strips were less sensitive to norepinephrine. It was found that desensitization to norepinephrine occurred following 10 min preincubation with 10(-6)m norepinephrine, and 20 min preincubation with 10(-7)m norepinephrine. The desensitization of supersensitive strips was significantly greater than in controls. Dopamine and isoproterenol desensitized both untreated and pretreated strips to norepinephrine. Phenylephrine desensitized strips to norepinephrine from 6-hydroxydopamine-pretreated rabbits. Acetylcholine and potassium did not desensitize strips to norepinephrine; however, acetylcholine desensitized strips to acetylcholine. It is concluded from these studies that norepinephrine desensitization in a specific alpha-adrenergic-related event.


Beta-adrenoceptor mediated inhibition by terbutaline of histamine effects on vascular permeability. 1 In guinea-pigs, previously given Evans blue dye intravenously, 13 to 18 intradermal injections of histamine, with or without other drugs, were made into the depilated trunk skin. Dye was then quantitatively extracted from each skin area and the measured absorbance values were used as a measure of vascular leakage of macromolecules. 2 Histamine (0.5 to 12 nmol) produced dose-related increased in vascular leakage. These were reduced by terbutaline (1 and 10 nmol) which produced a significant shift in the histamine dose-response lines to lower absorbance values. The effect of 0.1 nmol of terbutaline was significant only against doses of histamine of less than 2 nmol. 3 Propranolol (1 nmol and 10 nmol) antagonized the effects of terbutaline. Propranolol, at a dose of 10 nmol but not 1 nmol, itself reduced the responses to 1.5 nmol histamine. 4 We conclude that the inhibition by terbutaline of histamine-induced dye leakage in guinea-pig skin is mediated by stimulation of beta-adrenoceptors and its is suggested that this effect of terbutaline occurs directly on beta-receptors at the vascular leakage site.


The effects of a toxin isolated from Australian tiger snake (Notechis scutatus scutatus) venom on autonomic neuromuscular transmission. 1 The effects on mammalian autonomic neuromuscular transmission of a purified toxin from the crude venom of the Australian tiger snake, Notechis scutatus, have been investigated. 2 The toxin had no measurable effect on transmission in either the rat anococcygeus, the rat vas deferens, or the longitudinal muscle of the guinea-pig ileum. 3 The toxin induced a contraction of longitudinal smooth muscle of the ileum. The tissue relaxed in spite of the continued presence of the toxin, and remained insensitive to further doses. The contractile response was not mediated by either cholinergic or histaminergic mechanisms. 4 The toxin caused an inhibition of the response to field stimulation of the guinea-pig vas deferens and the guinea-pig seminal vesicle. The inhibition was spontaneously reversible, and the preparations remained insensitive to further doses fo the toxin. The effects of the toxin were not mediated by either prostaglandins or by noradrenaline. 5 The inhibitory effects of a variety of compounds known to act at presynaptic sites were also blocked by exposure to the toxin; inhibition caused by postsynaptic activity was unaffected by the toxin. 6 It is suggested that the toxin inhibits transmission in the guinea-pig vas deferens and seminal vesicle by a presynaptic mechanism. 7 The possibility that the mechanism involves phospholipase A2 activity is discussed.


Vasoactive intestinal polypeptide promotes cyclic adenosine 3',5'-monophosphate accumulation in guinea-pig trachea. Cyclic adenosine 3',5'-monophosphage (cyclic AMP) levels in guinea-pig tracheal rings increased on incubation with the vasoactive intestinal peptide (VIP). The effect was potentiated by the addition of theophylline. The results suggest that the tracheo-bronchial relaxant action of VIP may be mediated by stimulation of tracheal adenylyl cyclase.


An alternative to group homes.

An alternative to group homes. This paper describes a supported lodgings scheme as an alternative to group homes. It is pointed out that the County Council has a statutory duty to finance supported lodgings and that schizophrenics are ideally suited to such a scheme. Some short-stay, the majority of the 'new' non-demented long-stay, and a large number of the 'old' long-stay patients have been discharged by this means. After-care facilities were important, as nearly half attended the day centre and over one third were regularly visited in their lodgings by the community psychiatric nurses and social workers.


Cyclic AMP, adrenaline and noradrenaline in plasma during surgery. Plasma concentrations of cyclic AMP, adrenaline and noradrenaline were measured in 6 patients undergoing hysterectomy from before induction of anaesthesia to 6 h after skin incision. Noradrenaline did not vary significantly during the observation period, whereas cyclic AMP and adrenaline increased after skin incision. A significant correlation was found between plasma concentrations of cyclic AMP and adrenaline (r = 0.84, P less than 0.01), suggesting that the latter is responsible for the increase in plasma cyclic AMP which is observed in relation to surgical procedures. Peak concentrations of cyclic AMP and adrenaline were seen in the early postoperative phase. This indicates that the most pronounced acute endocrine stress response to surgery of moderate severity occurs after termination of anaesthesia.


Clinically silent gross hypergastrinaemia from a multiple hormone-secreting pancreatic apudoma. A patient is described who had a malignant pancreatic islet cell apudoma secreting corticotrophin (ACTH) and melanocyte-stimulating hormone (MSH), both of which were clinically active, and very large quantities of immunoreactive gastrins, which were biologically active but clinically silent (normal gastric acid secretion and no peptic ulceration). The presence of parietal cell antibodies, with no increase in the plasma concentrations of hormones which can inhibit gastric acid secretion (secretin, GIP and VIP), suggests that many of the of the parietal cells may have been blocked by the autoantibodies.


Reactions after pertussis vaccine: a manufacturer's experiences and difficulties since 1964. Pertussis vaccines vary in quality, safety, and efficacy according to the production strains of Bordetella pertussis, the method of manufacture, and quality control procedures. It is therefore not justifiable to combine information on the incidence, nature, and severity of reactions after all manufacturers' pertussis vaccines as if they were a single product. Attempts were made to collect information on all suspected cases of severe reactions that occurred after administration of about 15 million doses of Wellcome pertussis vaccines in the United Kingdom and Northern Ireland from 1964 to mid-1977. Altogether six deaths, six neurological reactions with sequelae, and 17 convulsions without sequelae were reported, but some were clearly not attributable to the vaccine, while, in other cases, the available information was inadequate for assessing the role of vaccination. Neurological disorders, similar to those reported in a few children after pertussis vaccination, occur unexpectedly in apparently healthy infants at the recommended age for immunisation, so chance association between vaccination and these events can be expected in some children. The Joint Committee on Vaccination and Immunisation has made several recommendations aimed at reducing severe reactions after pertussis vaccination. These include replacing plain vaccine with aluminium-adsorbed vaccine, but there is no clear evidence that the aluminium-adsorbed vaccine produces fewer reactions than the plain.There are difficulties enough in deciding the cause of events that occur after vaccination, since these reactions often occur naturally in children of vaccination age. The task is made even harder by the assumption that various manufacturers' vaccines are the same and the lack of information available to manufacturers about cases in which their vaccine has been implicated. Information on vaccines administered is entered on immunisation records cards; it should be used and referred to if reactions occur.


High-density and low-density lipoproteins and prevalence of vascular disease in diabetes mellitus. The prevalence of vascular disease among 154 diabetic patients was analysed in relation to the serum concentrations of individual lipoproteins. Overal the presence of vascular disease (59 cases) was positively associated with serum cholesterol and low-density-lipoprotein cholesterol but negatively associated with high-density-lipoprotein (HDL) cholesterol. The negative relation between HDL and vascular disease was not observed in all subgroups of diabetics. We conclude that there may be no overriding association between HDL and vascular disease in diabetics as proposed for some non-diabetic populations.


Effects of some conformationally restricted GABA analogues on GABA membrane binding and nerve ending transport. By using a series of aminocyclopentane- and aminocyclohexanecarboxylic acids, as well as some naturally occurring amino acids, it was possible to determine some aspects of the spatial topography of the GABA membrane binding and transport sites. The NA-independent GABA binding site was found to have a different spatial topography than the Na-dependent binding site in that trans-3-aminocyclopentanecarboxylic acid (trans-3-ACPC) was 7 times more potent than cis-3-ACPC to inhibit Na-independent binding, but only 1.6 times more potent to inhibit Na-dependent binding. The nerve ending GABA transport site was found to be similar to the Na-dependent GABA binding site in that it will accommodate both cis- and trans-3-ACPC. However, the transport site differs from the binding site in that cis-3-aminocyclohexanecarboxylic acid (cis-3-ACHC) is a potent inhibitor of transport but a weak inhibitor of binding. In addition to the differences in spatial characteristics, differences in the subcellular distribution of Na-independent and Na-dependent binding sites were observed. The former were found primarily in the nerve ending-mitochondrial fraction, while the latter were primarily found in the microsomal fraction.


Receptors for gamma-aminobutyric acid (GABA) on Aplysia neurons. Aplysia neurons show 5 different types of response (three excitatory and two inhibitory) to iontophoretic application of gamma-aminobutyric acid (GABA). Four of these are associated with a membrane conductance increase, but one is associated with a conductance decrease. The most common response is a fast hyperpolarization which reverses at about--58 mV and is sensitive to manipulation of external Cl- concentration, and thus is due to a specific increase in Cl- conductance. There is an infrequent, slower hyperpolarizing response which does not reverse above about--80 mV and is insensitive to external Cl-. This response appears to result from a conductance increase to K+. Two types of depolarizing responses are associated with conductance increases. These responses differ in their latency, duration and sensitivity to curare. The more frequent is relatively rapid (peak at 1-2 sec) and is depressed by curare at high concentrations. In other neurons, GABA causes a slower response, peaking at 6-10 sec, which is not curare-sensitive. Usually for both types of response, the voltage and conductance changes are completely abolished by perfusion with Na+-free seawater, and the responses cannot be reversed with depolarization. In other neurons such as L11, the response can be reversed with depolarization, and appears to result from a conductance increase to both Na+ and Cl-. In neuron R15, GABA causes a slow depolarizing response (peak at about 9 sec) which is associated with a decreased membrane conductance, probably to K+. The classical GABA antagonists, picrotoxin and bicuculline, block Cl- responses but no others, while the fast Na+ and Cl- responses are depressed by curare. Strychnine does not affect any GABA response. The multiplicity of GABA responses, the specificity of their organization and the fact that only some neurons have receptors for GABA, argue that GABA may have a role as a neurotransmitter in Aplysia. Furthermore, the existence of several types of excitatory GABA response suggests that GABA may function both as an inhibitory and excitatory neurotransmitter.