Proximal gastric vagotomy: interim results of a randomized controlled trial. In a randomized controlled trial 50 patients with duodenal ulcer treated by proximal gastric vagotomy (P.G.V.) without drainage were compared with 50 who underwent selective vagotomy and gastrojejunostomy. The clinical results were assessed in 99 patients one to four years after operation. Patients who had undergone P.G.V. had significantly less dumping, nausea, and bile vomiting and fared better in their overall clinical grading. The postoperative Visick grading of the 50 patients with P.G.V. was similar to that of 56 controls with no known gastrointestinal disease who had not undergone operation. The results obtained in the patients who had had P.G.V. without drainage were compared with those of a further group of 24 patients subjected of P.G.V. with gastrojejunostomy, and the better results obtained in the former group were thought to be due to elimination of the drainage procedure. The average follow-up period of the trial was just over two years, but there were no indications that the recurrent ulceration rate after P.G.V. would be any higher than after other types of vagotomy and drainage.


Report on injuries sustained by patients treated at the Birmingham General Hospital following the recent bomb explosions. As a result of recent bomb explosions a total of 82 patients were treated at the Birmingham General Hospital, 61 with minor injuries. Bomb injuries may be divided into three main groups due to the blast effect (such as blast lung and ruptured tympanic membranes), the flash (such as burns to the exposed part of the body), and shrapnel (which may cause a wide variety of injuries). The amount of warning of such explosions is usually minimal, and so the prepared accident schemes of most hospitals are inappropriate. If the disaster occurs outside normal working hours much responsibility initially falls on the resident staff. This report gives some idea of the type of injuries they are likely to see.


Myocardial infarction in young women with special reference to oral contraceptive practice. Sixty-three women discharged from hospital with a diagnosis of myocardial infarction and 189 control patients were studied. All were under 45 years of age at the time of admission. Current oral contraceptive use, heavy cigarette smoking, treated hypertension and diabetes, pre-eclamptic toxaemia, and obesity were all reported by, and type II hyperlipoproteinaemia was found more often in, patients with myocardial infarction than their controls. The relationship between myocardial infarction and oral contraceptives could not be explained in terms of an association between the use of these preparations and the other factors. The combined effect of the risk factors was clearly synergistic.


Oral contraceptives and death from myocardial infarction. We investigated 219 deaths from myocardial infarction in women under the age of 50. Their histories were compared with those of living age-matched controls selected from the same general practices. The frequency of use of oral contraceptives during the month before death was significantly greater in the group with infarction than during the corresponding month in the control group and the average duration of use was longer. No information of cigarette smoking was available but the proportion of women being treated for hypertension or diabetes was greater among those who died than among the controls. This did not alter the overall conclusion that the risk of fatal myocardial infarction was greater in the women using oral contraceptives, particularly in the older age groups.


Diarrhoeae in thyroid medullary carcinoma: role of prostaglandins and therapeutic effect of nutmeg. In a patient with medullary carcinoma of the thyroid with pulmonary metastases who presented with diarrhoea and steatorrhoea large amounts of prostaglandin-like material were present in peripheral blood, and some was extracted from the tumour. The diarrhoea which persisted after thyroidectomy responded to treatment with nutmeg.


Midbrain structures controlling vocalization in the domestic chick. Bilateral lesions confined to the anterior intercollicular area (ICo) produce muting in domestic chicks. Calling is lost both in open field test, and to novel stimuli which evoke pecking. Lesions of the medial edge of the ICo also mute, perhaps by cutting connections with the central mesencephatic grey. Partial destruction of the ICo allows full intensity calling but with greatly increased latency. If an area ventral to the ICo is also destroyed, one type of calling reappears: it is still not readily evoked by particular stimuli which are effective in normal chicks but it often occurs during locomotion. Crowing survives ICo lesions and presumably depends on a different route to lower centres. Totally deaf birds show quite normal calling, so that hearing deficits are probably not involved in the muting. Finally, mammalian homologues of the ICo and its connections are identified.


Changes in visual responsiveness following intercollicular lesions and their effects on avoidance and attack. In the normal chick, conspicuous visual stimuli induce targetting and pecking together, with vocalization. All three are abolished by lesion of the intercollicular area (ICo) or of connections passing through its medial margin. After such lesions, chicks also cease to treat significant visual stimuli as if they were startling and exciting, and may delay response as a result. However, they are still able to recognise, orient accurately to, and respond appropriately to, a variety of complex visual stimuli (e.g. food grains, copulation object). In addition, they are little affected by strange surroundings. Lesion evidence suggests the mammalian subcollicular area to have similar functions to the ICo and to be homologous with it. A route (present in bird), which is well-known in mammals for its association with threat, defense and escape evoked by strange and frightening objects (amygdala-diencephalic periventricular system-central mesencephalic grey, A-DPS-CMG) is stimuli via the 2 ICo (subcollicular area). Two different mechanisms may be involved caudal to the ICo. One consists of tectal afferents which might modulate the evocation of targetting, pecking and other responses via the tectum. The other is the predorsal system of tectal efferents which may mediate such responses. Classical syndromes of tameness and unresponsiveness produced by various interruptions of the A-DPS-CMG route may depend on interruption of connections to these midbrain mechanisms. Attack is depressed by ICo lesions as one aspect of reduced responsiveness to conspicuous and startling visual stimuli. Avoidance, which is apparently mediated by a separate system, much as in Anura, is facilitated.


Macromolecular complexes produced by 1,3-propanesultone. 1,3-Propanesultone produces in vitro complexes between DNA and DNA, DNA and RNA, RNA, and RNA, DNA and proteins, and possibly RNA and proteins. All these interactions were detected with a variety of independent analytical techniques. Increased attachment of the 1,3-propanesultone-treated DNA to the microsomal membrane and enhanced adsorption to and penetration into Ehrlich ascites tumor cells were also observed. 1,3-Propanesultone was the second small-ring alkylating carcinogen studied in this laboratory that was found to produce such macromolecular complexes. The possibility that these highly diverse structures may be involved in chemical mutagenesis and tumorigenesis is considered.


Factors influencing the survival of rat brain tumor cells after in vitro treatment with 1,3-bis(2-chloroethyl)-1-nitrosourea. The shape of dose-response curves obtained for asynchronous, exponentially growing 9L rat brain tumor cells treated in vitro with 1,3-bis(2-chloroethyl)-1-nitrosourea changed as a function of the drug exposure time. For short treatment times (less than 1 hr), the dose-response curves had shoulders, indicating that the cells may accumulate sublethal damage; however, after longer treatments (greater than 1 hr), little if any shoulder was apparent. The slope of the exponential portion of the dose-response curve increased progressively with treatment periods from 15 min to 2 hr. Longer exposure times (up to 24 hr) produced no further changes in the cell-kill kinetics. Cell survival was directly related to the BCNU exposure dose [o integral t Co(t)dt] and to the amount of bound BCNU per cell. Extrapolation of the curves for these two variables indicated that some BCNU damage accumulates before death occurs. The amount of serum and cell products available in the medium to bind BCNU affected the level of survival; however, there was no evidence that extracellular spontaneous breakdown products or chemical transformation products were involved in the cell-killing mechanism.


Stem-cell survival and tumor control in the Lewis lung carcinoma. The stem-cell response of the Lewis lung carcinoma to single doses of cyclophosphamide has been studied by three assay techniques: in vitro colony formation, lung colony formation, and the end-point dilution assay. These three techniques have given comparable results, and the end-point dilution results showed that 50 percent takes could be achieved with as few as 1 to 3 cells. Studies have been made of the growth of small i.m. implants and of the time following implantation at which they could be eradicated by cyclophosphamide. The results were compared with the curability that would be expected on the basis of cell survival studies. It was found that older (and therefore larger) implants were cured than might have been expected. It seems unlikely that this discrepancy was due to additional cell kill caused by an immune response. An alternative explanation, that the surviving fraction following a dose of cyclophosphamide was lower in small implants than in larger i.m. tumors, was supported by studies of cell survival in dissectable lung colonies.


Binding of daunomycin to DNA and the inhibition of RNA and DNA synthesis. With synchronized tissue culture cells (L929), daunomycin had the greatest inhibitory effect on cell growth when the drug was administered during the later stages of cell division (late S, G2, and M). The level of binding of daunomycin to DNA was not found to be influenced by the phase of the cell cycle. The highest level of radioactivity from [eH]-daunomycin was bound to DNA of the heterochromatin fraction. Both RNA and DNA syntheses were inhibited in isolated enzyme systems when daunomycin-treated DNA, from which the unbound drug was removed by passage through Sephadex column, was used. DNA polymerase was reduced to one-fifth of the control activity, while that of RNA polymerase was reduced to one-half. Similar experiments with daunomycin-treated RNA and DNA polymerase preparations showed that the drug had no effect on the activities of the enzymes per se. Hence, the reduction of RNA and DNA polymerase activities could be accounted for by the loss of template activity of the drug-treated DNA. Daunomycin caused by a marked drop in the formation of a complex between RNA polymerase and DNA, indicating that the binding of daunomycin to DNA may give rise to steric hindrance effects that interfere with the association of the template to RNA polymerase enzyme. Sedimentation profile in alkaline sucrose density gradient of DNA that had been treated with daunomycin showed that no change in the molecular weight could be demonstrated.


The role of cell division in the malignant transformation of mouse cells treated with 3-methylcholanthrene. The requirement for cell division in the malignant transformation of A31-714 cells, a subclone derived from BALB/3T3, by 3-methylcholanthrene was investigated using the property of the high susceptibility of this clone to density-dependent inhibition of cell growth. Treatment with 3-methylcholanthrene did not induce transformation in a nongrowing population. However, the cells treated with the  arcinogen in a nongrowing state showed a high transformation frequency near maximum level when they were returned to the growing state soon after treatment. About four cell generations were found to be necessary for the development of cell transformation after treatment with 3-methylcholanthrene. Cells that were kept in a nongrowing state after carcinogen treatment rapidly lost their ability to express transformation even when they were subsequently returned to a growing state. On the other hand, the cells that were allowed one cell division soon after carcinogen treatment retained their ability to produce transformed foci even after being kept in the nongrowing state thereafter. These results suggest that one cell generation is required for the fixation of transformation and that several additional cell generations are required for the expression of the transformed state.


Investigation of specific antigens in prostatic cancer. Rabbits immunized with normal human prostatic homogenates have been shown to develop antibodies specific for one form of prostatic acid phosphatase and a protein antigen which does not appear to possess enzymatic activity. Antibodies were also developed against several other proteins, esterase, two aminopeptidases, and a second form of acid phosphatase. Extensive absorption of the antisera with gluteraldehydepolymerized serum and kidney homogenate removed all antibodies except those which react with the specific acid phosphatase and the specific protein. These two antigens have been separated from each other by ion-exchange chromatography. The partially purified acid phosphatase has been used to produce antiserum employed for the detection of prostatic cancer in bone marrow aspirates.


Identity, release, and binding of mitochondrial-bound hexokinases in mammary glands and adenocarcinomas of lactating mice. The mitochondrial-bound hexokinases (adenosine triphosphate:D-hexose 6-phosphotransferase) of mammary adenocarcinoma and of normal gland were compared in lactating C3H mice. Treatment of mitochondria isolated from both the normal and neoplastic tissue with 0.5 m NaCl or 0.1 mM glucose 0-phosphate effected the release of about 50% of the bound hexokinase. In the presence of magnesium ion, enzyme from either source attached to mitochondria from either tissue and in all combinations to the same extent. Identification of the isoenzyme complement in the mitochondrial extract by diethylaminoethylcellulose chromatography revealed only types I and II. In the tumor, the hexokinase activity in both the cytosol and the fraction solubilized from mitochondria was predominantly in the form of type I ( 60%). In contrast, the activity released from mitochondria isolated from normal gland was predominately type II, while the cytosol contained almost equivalent amounts of types I and II. While this difference does not explain differences in glucose utilization between the normal and neoplastic tissue, it may provide a means of distinguishing between the two.


A simple radioimmunoassay of human placental alkaline phosphatase (Regan isoenzyme) using specific antibody polymers. Rabbit antiserum against highly purified high-molecular-weight B-variant of human placental alkaline phosphatase (M.W. 200,000) was rendered monospecific by absorption with polymerized pooled male serum proteins; the absorbed antiserum was then polymerized with ethyl chloroformate and used in radioimmunoassay as a stable solid-phase immunoabsorbent. Homogeneous preparation of the enzyme, with a specific activity of 477 mumoles phenol per mg per min, was also obtained by absorbing the chromatographically purified enzyme with polymerized rabbit antiserum directed to whole human serum proteins; the pure enzyme was then labeled with 125-I as the tracer retaining at least 80% of its antigenicity. Only a minute quantity of the polymerized antibody particles is required for each assay in admixture with the labeled and unlabeled enzyme. By adding a small amount of starch-gel particles before low-speed centrifugation, complete phase separation was achieved. The radioimmunoassay could detect 0.4 to 0.8 ng enzyme protein per tube, which is comparable to the sensitivity achieved by enzymic assays. However, radioimmunoassay is advantageous over the enzymic assay in being direct, specific (no interference by the nonplacental-type alkaline posphatase), and capable of detecting both catalytically active and inactive forms of the enzyme. Native variants of placental-type alkaline phosphatase including Regan isoenzyme and Nagao isoenzyme (D-phenotype of normal placental alkaline phosphatase), could thus be directly determined by this procedure in the clinical specimens.


Cellular differentiation of rat uterine adenocarcinoma cells by progesterone in vitro. Transplantable cloned HTP/Cl culture was a stable line derived from a rat uterine adenocarcinoma that was induced by 7,12-dimethylbenz(a)anthracene in vivo and did not display density-dependent inhibition of growth. This HTP/Cl culture easily adapted to grow in a culture medium containing progesterone, 8 mug/ml. As compared with HTP/Cl culture, HTP/Cl/P8 culture grown in the presence of progesterone was contact inhibited, and a cellular differentiation was observed in the tumor tissues that developed after inoculation of cells. These actions of progesterone on uterine adenocarcinoma cells were completely reversible on removal of the hormone in vitro. These results appeared to indicate that progesterone was involved in the regulation of both cellular proliferation and differentiation. The possible mechanisms of the regulation of rat uterine adenocarcinoma cells by progesterone are discussed in relation to cellular levels of cyclic adenosine 3':5'-monophosphate in vitro.


The synthesis of Forssman glycolipid in clones of nil 2 hamsters fibroblasts grown in monolayer or spinner culture. The amount of Forssman glycolipid (GL-5) was investigated in two clones of Nil hamster cells grown either in monolayer or in spinner culture. GL-5 assayed by the complement-fixation inhibition test increased with increasing cell density in the monolayer. However, cells grown in spinner cultures failed to show the density-dependent response in both clones examined. Cells transformed by hamster sarcoma virus did not show the density-dependent increase of GL-5, even in cells grown in monolayer. The effect of transfer from confluent to sparse cultures on the amount and the synthesis of GL-5 was also examined. It is suggested that the GL-5 that accumulates in cells during confluency is diluted into the daughter cells and that the decrease of the Forssman lipid does not precede cell division.


Further evidence for a basis of selective activity and relative responsiveness during antifolate therapy of murine tumors. A greater persistence of unbound (exchangeable) drug in tumor cells versus drug-limiting normal tissue (proliferating epithelium of small intestine) correlates with the therapeutic effects of various antifolates against a group of murine tumors. After approsimate equimolar doses (3 mg/kg i.p.) of methotrexate (MTX) methasquin (MQ), aminopterin, and N-([2,4-diamino-5-chloro-6-quinazolinyl) methyl]-amino)benzol)-L-glutamate (5-Cl-deaza-AM), total accumulation in small intestine was vie- to eight-fold greater than the dihydrofolate reductase content. Free drug persisted for less than 4 hr (MTX), 16 hr (MQ), 30 hr (aminopterin),and 48 hr (5-Cl-deaza-AM). Overall drug accumulation in L121O cells was greater (12- to 40-fold enzyme level), and drug persistence above enzyme level was more prolonged in the case of MTX (24 hr),MQ (32 hr), and 5-Cl-deaza-AM (greater than 48 hr). Persistence of aminopterin was similar to that seen in small intestine. After the same dose of each drug s.c., the results were similar in small intestine. In L121o cells, however, the total drug accumulation was much lower, but the relative persistence of each was similar to that seen after an i.p. dose. After a single optimal therapeutic dose (3, 0.75, 0.3, and 0.1 mg/kg i.p. for MTX, MQ, aminopterin, and 5-Cl-deaza-AM every other day), accumulation of each drug in for no more than 4 hr. In L121O cells, maximal accumulation of each drug also varied, but persistence differed in accordance with the relative therapeutic effectiveness of each (9 hr for 5-Cl-deaza-AM, 12 hr for aminopterin, and more than 20 hr for MTX amd MQ).


Occurrence of hemangiosarcomas in beagles with internally deposited radionuclides. In a series of related experiments to evaluate the relative toxicity of inhaled radionuclides, beagles were exposed to aerosols containing relatively soluble (chloride) or relatively insoluble (fused clay) forms of 144-Ce and 90Sr. With the solubled 144-CeCl3, significant radiation doses were delivered to the lungs, liver, and skeleton whereas, after 90-SrCl2 exposure, the radiation dose was delivered predominantly to the skeleton. In dogs exposed to 144-Ce and 90-Sr in fused clay particles, radiation doses were delivered mostly to the lungs and tracheobronchial lymph nodes. In most dogs dying within 2 years after exposure, deaths were attributable to nonneoplastic radiation-induced lesions in the target organ systems. At later times after exposure, neoplasms were the major cause of death, again occurring mostly in target organs or the adjacent tissues. Lung liver, and bone-related neoplasms, including five hepatic hemangiosarcomas, developed after 144-CeCl3 exposure. Among the bone-related sarcomas seen in dogs exposed to 144-CeC3 or 90-SrC2, the incidence of hemangiosarcomas was over 40%. Among the 20 dogs dying with pulmonary neoplasms after exposure to 144-Ce or 90Sr in fused clay particles, all had hemangiosarcomas and several also had other neoplasms. This high after exposure and differs from results in other laboratories where beagles have been exposed to both alpha and beta-emitting radionuclides.


Absence of specificity in inhibition of DNA repair replication by DNA-binding agents, cocarcinogens, and steroids in human cells. Although many chemicals, including cocarcinogens, DNA-binding agents, and steroids, inhibit repair replication of ultraviolet-induced damage to DNA in human lymphocytes and proliferating cells in culture, none of these chemicals is specific. Our results show that all the chemicals than they inhibit repair replication. There is thus no cocarcinogens are specific inhibitors of DNA repair or that any of the chemicals studied might be useful adjuncts to tumor therapy merely because of specific inhibition of radiation repair mechanisms.


Alteration of the syncytium-forming property of XC cells by productive Moloney leukemia virus infection. The effect of productive murine leukemia virus (MuLV) infection of the syncytium-forming property of XC cells was studied MuLV-Moloney-infected XC cells, designated XC(M), initially went through a period of spontaneous syncytium formation. The syncytia then disappeared, and XC(M) cells continued to propagate and produce both infectious MuLV and MuLV group-specific antigens. However, XC(M) cells became refractory to syncytium formation that was induced by Kirsten, Friend, Rauscher, and Moloney strains of murine oncornaviruses. These data suggest that XC(M) cells lost their syncytium-forming ability as a result of productive MuLV infection.


Localization of the beta chain of human chorionic gonadotropin on human tumor cells and placental cells. With the use of peroxidase-labeled antibody to the beta chain of human chorionic gonadotropin, sections of ten human malignant tumors were found to react with this antibody. It is postulated that both selective host immunosuppression by tumors and selective maternal immunosuppression by fetal tissues may be mediated by human chorionic gonadotropin.


Growth of rhabdomyosarcoma colonies from pleural fluid. Pleural fluid from a child previously treated for rhabdomyosarcoma produced colonies in vitro. Cells from these colonies appeared to have the light and electron microscopic appearance of rhabdomyosarcoma cells. In this case, the malignant nature of the effusion had been suspected because of the patient's previous history; however, this technique may prove useful in the diagnosis of effusion of unknown etiology.


On the water-soluble heterogalactan from the fruit bodies of Lentinus edodes. The chemical structure of a heterogalactan isolated from the trichloracetic acid extract of the fruit bodies of Lentinus edodes is reported. It consists of a main chain of (1 yields 6)-linked alpha-D-galactopyranose residues, part of which are substituted in the 2-position either with single L-fucopyranose or D-mannopyranose residues. However, there is a possible alternative structure of a branched D-galactan in which most of the side-chains are terminated with L-fucose or D-mannose residues.


The synthesis, turnover, and artificial restoration of a major cell surface glycoprotein. A large glycoprotein (molecular weight equals 220,000 daltons) can be isolated from the surfaces of chick embryo fibroblasts. Such a protein had been shown by others to be present on normal cells in vitro, but missing from transformed variants of these cells, suggesting a possible role in growth control. We have estimated the turnover rate of this cell surface protein (CSP) and show that it is not more rapid than that of other cell proteins. We have also examined the resynthesis of CSP after removal from cells by trypsinization. This restoration is gradual, requiring more than 24 hr, and is affected by cell density. Restoration can be blocked by cycloheximide. Using cycloheximide to maintain cells depleted of CSP, we have demonstrated that isolated CSP can be progressively reabsorbed at 37 degrees C onto these denuded cell surfaces, and that this adsorption is not observed at 4 degrees C. This information on the turnover, depletion, and restoration of CSP may provide a means of directly testing its function.


Murine mammary gland RNA directed synthesis of casein in a heterologous cell-free protein synthesis system. A cell-free protein synthesis system derived from Ehrlich ascites tumor cell ribosomes (S30) plus rabbit reticulocyte tRNA was developed and the activity of the system was dependent on rabbit reticulocyte ribosomal salt (0.5 M KC1) wash factors, The exogenous mRNAs from BALB/c mouse liver and the mammary gland were translated with a high efficiency in this heterologous cell-free system. Furthermore, the RNA from the lactating mammary gland faithfully directed the synthesis of casein. The presence of mouse casein in the reaction product was identified by radioimmunoprecipitation with mouse casein antiserum, co-electrophoresis of the reaction product and mouse casein the urea-polyacrylamide gel and by electrophoresis in sodium dodecyl sulfate (SDS) polyacrylamide gel. The major portion of the lactating mammary gland RNA directed synthesis of the milk protein in the cell-free system appeared to be analogous to alphas casein,


Serotonin transport by cultured bovine aortic endothelium. Endothelial cells isolated from bovine aortas without prior treatment with enzymes were cultured in RPMI 1640 medium containing 17% fetal calf serum and antibiotics. The endothelial cells at confluency (7 days) were similar to endothelium in situ or to freshly isolated endothelial cells from blood vessels as seen by light, scanning, and electron microscopy. Cultured and freshly isolated indothelial cells exposed to labeled serotonin, even in the presence of iproniazid (5 x 10-4M), took up approximately 125 and 250 pmoles 14C-serotonin/mg protein, respectively, in 3 hours. Imipramine (10-4M) reduced uptake for both cell groups. Cold (4 degrees C) and metabolic inhibitors sharply reduced serotonin uptake by both freshly isolated and cultured endothelial cells. Ouabain (10-5M) almost completely blocked serotonin transport. Six analogues of serotonin at concentrations ten times above experimental serotonin concentrations did not affect serotonin transport in the cultured endothelial cells but did reduce it in the freshly isolated endothelial cells by 50%. The data on transport suggest that serotonin uptake is not unique to pulmonary endothelium, as has been suggested previously. In addition, using cultured indothelial cells to study serotonin transport is compatible with using other serotonin model systems such as platelets, lung, or brain. Lastly, serotonin uptake by endothelial cells may involve an active transport mechanism similar to that described for the pulmonary circulation, platelets, and insect salivary glands.


Changes in renin, water balance, and sodium balance during development of high blood pressure in genetically hypertensive rats. In genetically hypertensive rats of the Milan hypertensive strain (MHS) and normotensive rats (NR) developed from the same Wistar stock colony, blood pressure (BP), plasma renin activity (PRA), sodium balance, and water balance were measured from the time of weaning to the seventh week postweaning in three separate but essentially identical experiments. In a fourth experiment, total and extracellular water, total sodium, and exchangeable sodium were measured in MHS and NR at 24 and 130 days of age. Although the time course of changes varied slightly between experiments, BP of both NR and MHS rose until the second and third weeks postweaning, at which time BP in MHS was 40 to 50 mm Hg higher than in NR. PRA in MHS was one-half that of NR at weaning. Increasing BP was accompanied by falling PRA in both, and PRA was not significantly different when stable, adult blood pressure was reached. Urinary volume in MHS was 50% to 100% greater (P less than 0.001) than in NR at weaning and for a few days after. Sodium was retained to a greater extent by MHS during the period when the blood pressure difference develops, from weaning to the fourth week postweaning. This sodium retention (MHS = 97.0 plus or minus 10.3, NR = 65.2 plus or minus 6.8 SE mu-Eq Na retained/g body weight gain; P less than 0.005) is the result of significantly lower urinary excretion of dietary sodium by MHS. A causative role for the kidney is suggested in the established of high blood pressure in MHS.


Echocardiographic features of subpulmonic obstruction in dextro-transposition of the great vessels. Eleven patients with dextro-transposition of the great vessels (TGV) and subpulmonic (left ventriclar outflow) obstruction documented by cardiac catheterization and angiography were studied by echocardiography. Echocardiographic features were which characterized the outflow obstruction included a) prolonged diastolic apposition of the mitral valve with the ventricular septum (10 patients), b) smaller width of the pulmonary artery as compared to the aortic root (10 cases), and c) narrowing of the subpulmonic area demonstrated by beam anglation studies through the planes of the mitral valve and the pulmonary artery (five cases). In addition, four patients showed abnormal systolic anterior movements (SAMs) of the anterior mitral leaflet resembling those observed in idiopathic hypertrophic subaortic stenosis. The SAM'S WERE LARGE IN THREE PATIENTS AND MAY HAVE ACCENTUATED THE OUTFLOW OBSTRUCTION. Additional evidence for this was provided by the movement of the pulmonary valve towards closure with the onset of the mitral SAM'S. These features were uncommon or absent in a comparison group of 17 patients with TGV and no outflow obstruction. Echocardiography appears to be useful in the diagnosis of subpulmonic muscular obstruction in TGV. Demonstration of definite mitral SAM'S IN SOME PATIENTS SUGGEST THE PRESSENCE OF ASSOCIATED FUNCTIONAL OBSTRUCTION OF THE TYPE SEEN IN IDIOPATHIC HYPERTROPHIC SUBAORTIC STENOSIS.


Operative treatment in hypertrophic subaortic stenosis. Techniques, and the results of pre and postoperative assessments in 83 patients. The results of operative treatment in 83 patients with idiopathic hypertrophic subaortic stenosis (IHSS) are described. Most patients with the disease are asymptomatic, or derive satisfactory symptomatic improvement from nonoperative therapy: administration of propranolol, exerice limitation, control of arrhythmia, etc. Operation is required, however, in 10-15% of patients, those who remain severely symptomatic after nonoperative treatment or who become refractory to it. Operation relieves symptoms in IHSS by relieving obstruction to left ventricular outflow, and for a patient to be considered an operative candidate severe obstruction must be documented at left heart catheterization either under resting conditions or after provocative interventions. All 83 patients were severely incapacitated--58 in Class III and 24 in Class IV. Seventy had obstruction at rest (average gradient 96 mm Hg), and 13 had only provocable obstruction. At operation the hypertrophic interventricular septum was exposed via an aortotomy, and a vertical bar of muscle was resected between parallel myotomy incisions. There were six operative deaths (7%); no patient has died since 1970. Seven patients have died late after operation, five of them from causes unrelated to their heart disease or the operation. All surviving patients describe symptomatic improvement. Fifty-two patients with obstruction at rest preoperatively (average gradient 95 mm Hg) have been studied postoperatively: no resting gradient was evident in 47, while in the remaining five the gradient was less than 25 mm Hg. Recurrence  of obstruction has never been observed at late  catheterization (21 pts) or late echocardiographic examination (37 pts). Obstruction could not be provoked postoperatively in ten of the 11 patients who had large gradients only with the Valsalva maneuver or isoproterenol administration preoperatively. Obstructed and provocable obstructed patients had similar symptomatic improvement after operation. A variety of rhythm and conduction abnormalities were observed both pre and postoperatively, and these are described in detail. The results of operation in these 83 patients with IHSS  demonstrate that gratifying symptomatic and hemodynamic improvement uniformly follows left ventriculomyotomy and myectomy. Relief of obstruction and amelioration of symptoms have proved to be long-lasting during postoperative observation  periods extending to 14 years. Continued application of the operative procedure in properly selected patients appears to be indicated.


Standards for total serum protein assays--a collaborative study. We have studied the standardization of total serum protein assay with the biuret reaction. Standard solutions were prepared from lyophilized preparations of human serum albumin and bovine serum albumin, with corrections made for volatile material and ash contents. These solutions and a solution of crystalline albumin standard were analyzed with a new stable biuret reagent, to establish absorptivity values (values for the absorbance of a 1 g/liter final reaction mixture). The mean values obtained were 0.302, 0.292, and 0.290 for human serum albumin, bovine serum albumin, and the crystalline albumin, respectively. We believe that the established absorptivity value will improve the accuracy of serum protein determinations. We studied the linearity of the relation between color produced and protein concentration, with use of the solutions described above and a serum pool. The color adheres to Beer's law up to the highest concentration tested: 3 g/liter for HSA and BSA, and 2.8 g/liter for serum in the final reaction mixture. The new biuret reagent has been stable for one year at room temperature. We recommend the use of bovine serum albumin as a primary standard for serum protein assays. It is inexpensive, easily available, and exhibits the best linearity in the biuret reaction.


A rapid 5 hour radioimmunoassay of progesterone and oestradiol in human plasma. A rapid 5 h radioimmunoassay method for the determination of progesterone and oestradiol in the plasma of non-pregnant women is described. Due to the high specificity of the antisera used, it is possible to perform the radioimmunoassay directly on the ether extracts of plasma, without employing chromatographic purification of the steroids. Evidence is presented indicating that the rapid assay is almost as reliable as the previously described radioimmunoassay method which involves chromatography. The within-assay and between-assay coefficients of variation in the progesterone assay were 7-74 and 14-9 and in the oestradiol assay 7-36 and 18-1 respectively. Comparisons between increasing doses of authentic hormone and endogenous hormone extracted from plasma indicated no deviation from parallelism. Progesterone and oestradiol were assayed in 300 plasma samples by the rapid method and by the method involving chromatography. The slopes obtained by a regression analysis were close to unity for both progesterone and oestradiol (1-04 and 1-06, respectively), the y-intercepts were - 0-21 and 0-16 and the correlation coefficients 0-98 and 0-88, respectively. When the data obtained by both techniques in fourteen menstrual cycles were compared, the results were practically identical. In ten repeated studies conducted by four investigators it was shown that two workers can complete the assay of both progesterone and oestradiol in twenty-five plasma samples in duplicates with 5 h. The same time required for the assay of either progesterone or oestradiol in twenty-five duplicates by one worker.


Visual attention to size and number of pattern details by term and preterm infants during the first six months. 2 dimensions inversely varied in previous complexity studies using checker-boards were varied independently. Patterns with more elements, angles, and information as well as patterns with larger elements received longer fixation at all ages, but initially prepotent size preferences decreased with age while number preferences became stronger. Further analysis showed the inadequency of contour length as the stimulus determinant for, or of increasing "optimal complexity" with age as the interpretation of, these previous findings. Term-preterm comparisons at 5-week intervals proved changes were a function of prenatal plus postnatal development rather than age from birth alone.


[Galactography of the female breast (author's transl)]. Breast lesions were diagnosed in 1518 patients among 8432 who had had mammography [ 19%].  In 324 there was the characteristic appearance of "secretory disease" .  Additional uni- or bilateral galactography was performed in 458 womenbecause of breast secretion.  A normal milkduct system was found in 29.6% of them; ductalectasia with a lumen above 3 mm diameter was present in 14.6%.  The most common cause ofsecretions was small cysts as part of a small-cystic mammary fibrosis or fibrocysticdisease of the breast [ 32.6%].  Intraductal proliferation was present in 21% of allgalactograms, occult carcinoma in 3 [ 0.7%].  It would seem that a secreting breast isnot as common an early symptom of mammary carcinoma as sometimes stated.  The most commoncomplication of galactography was mastitis [ more or less severe], almost always re-sponding to oxyphenbutazone administration.


[Acute renal failure after intracranial injury in three children: analysis of possible causes (author's transl)]. In three children (aged 6-8 years)acute renal failure developed three to five days after head injury. After emergency treatment and measures to prevent cerebral oedema, two of the children were given simultaneously dextran, cefalotin, gentaminic and beta-aescin. The dosage of these drugs was much higher than the usual upper limbs, beta-aescin in doses as high as 10 to 20 fold above recommended maximum. There was no proven connection between the drugs and their dosage and renal failure, because many factors were involved. However, in intensive treatment for children an exact control of drug dosage essential because toxic side effects of different drugs can be potentiated. All three children were haemodialysed, renal function recovered and all were discharged without organic sequelae.


The semen of fertile men: statistical analysis of 1300 men. The prevasectomy semen analyses of 1300 men who had fathered at least two children were studied. The ages of the men ranged from 23 to 64 years, with a mean age of 39.6 years; 84% of the men were Caucasian, 5% were black, and 5% were of other ethnic groups; 32% of the men were Roman Catholic, 26% Jewish, 22% Protestant, and 11% professed no religion; in 9% religion was not recorded. The number of children ranged from 2 (48%) and 3 (30.6%) to 10 (0.1%). The volume of the ejaculate ranged from 0.1 to 11.0 ml, with a mean of 3.2 ml plus or minus 1.4 SD. Sperm density ranged from 1.5 to 375 million/ml, with an arithmetic mean of 79 million/ml plus or minus 57 SD; however, the logarithmic or geometric mean was chosen as being more representative of the asymmetric distribution under study. The geometric mean was 65 million/ml, with standard deviation limits of 30 to 142 million/ml, figures that correspond to the 16th and 84th percentiles. The percentage of motile sperm ranged from 5 to 95%, with a mean of 65% plus or minus 22 SD. The grade of sperm motility, according to MacLeod and Gold's classification (Fertil Steril 2:187, 1951) of 0 to 4, ranged from 1 to 4, with a mean grade of 3. There were 25 men (2%) with a sperm count below 10 million/ml and 26 (2%) in whom the percentage of motile sperm was less than 20%. Polyzoospermy (count greater than 250 million/ml) was found in 15 men (1.2%); one or more spontaneous abortions had occurred in six of the spouses of these men (40%), whereas the over-all spontaneous abortion rate was 7% and the spontaneous abortion rate for wives of men with sperm counts below 10 million/ml was 4%. A positive relationship was found between sperm density and percentage of motile sperm; the relationship between motility and the logarithm of sperm density was represented by a straight line (a 10-fold increase in sperm count was accompanied by an increase of 20.4% in sperm motility); the slope of the line was highly significant, although the correlation coefficient was weak (0.27). A similarly positive relationship was found between sperm density and grade of motility; however, the correlation coefficient was also weak (0.22). The relationship between the grade of motility and the proportion of motile sperm also positive, was reasonably large (0.70), although its significance is questionable because of the crudeness of the scale utilized to assess the grade of sperm motility. A comparison of the findings with those of similar studies was made.


Ultrastructural observations on polyspermic penetration of zona pellucida-free human oocytes inseminated in vitro. An ultrastructural analysis of polyspermic human ova, obtained by insemination in vitro of zona pellucida-free oocytes, has demonstrated the following: 1. Sperm with an intact or reacted acrosome fuse equally well with the oocyte, and the sperm head is incorporated first. Sperm flagellum entry into the ooplasm is accomplished by sliding, while the sperm plasma membrane is being incorporated into the vitelline membrane. 2. Dismantling of the sperm flagellum within the ooplasm rapidly occurs by dispersion and disintegration successively of the mitochondria, the outer dense fibers of the midpiece, and the fibrous sheath of the principal piece. The axial filament complex persists much longer, and can be demonstrated by electron microscopy for 24 hours or more postinsemination. 3. The mode of assembly of the male pronucleus membrane and the incorporation of maternal chromosomes into the female pronucleus. 4. The vital role of the zona pellucida, which appears to be the essential, if not the unique, site of the block to polyspermy.


Estrogen secretion and its relation to corpus luteum formation in monkeys (Macaca fascicularis) given melatonin. Daily injection of 10 mg of melatonin from the 7th through the 17th day of the menstrual cycle in the crab-eating monkey delays the appearance of increased secretion of progesterone or prevents it altogether. In the cycles following melatonin treatment, the time in the cycle when the preovulatory estrogen rise occurs is more variable and the frequency of a second estrogen peak increases.


Angiosarcoma of the liver associated with Fowler's solution (potassium arsenite). A patient with an angiosarcoma of the liver associated with the chronic ingestion of Fowler's solution (potassium arsenite) is described. The patient's clinical course was characterized by upper gastrointestinal hemorrhage, recurrent hemoperitoneum, hepatic failure, and, subsequently, the appearance of an angiosarcoma of the skin. Selective angiography demonstrated features consistent with both hepatoma and cavernous hemangioma. The hepatic toxicity and the carcinogenicity of arsenic are reviewed, with particular reference to Fowler's solution, which previously has been widely used for the treatment of psoriasis. The long latency period for the development of the malignancies is emphasized.


[Severe incontinence of urine II degrees and its surgical treatment. (Indications for typical operations in cases of incontinence of urine and their results) (author's transl)]. Various surgical methods for the treatment of severe incontinence of urine II degrees can be used. The aim of the present study was to apply primarily in 255 cases of severe stress incontinence. II degrees the specific surgical procedure for treatment: 123 cases of cysto-rectocele repair including vaginal hysterectomy, 71 cases of puborectalis repair, 43 cases of urethrovesicosuspension operation with or without abdominal/vaginal supplementary procedures, till 1970 12 cases of combined operations and finally beginning in 1973 6 cases of dura-sling operation. Indications and principles of surgical intervention are described according to clinical intern procedures. Puborectalis repair (Franz operation) and pubococzygeus repair (Ingelman-Sundberg operation) in cases of missing prolapse combined with severe incontinence gave rather good results. The basis for optimum results after operative treatment of patients with stress incontinence is a detailed pre-operative diagnosis; The most specific operative procedure from the beginning seems to us more recommendable than routine cysto- rectocele repair including a second more specified operation in cases of relapse.


[Induction of ovulation with Epimestrol (author's transl)]. Epimestrol 5 mg. daily for 10 days was administered in 119 treatment cycles to 21 women with normal gonadotropins and 33 ovulations were induced. Of the 21 patients, 7 had secondary amenorrhea, 10 had anovulatory oligomenorrhea and 4 had ovulatory oligomenorrhea. 17 patients desired pregnancies and 6 became pregnant. There was 1 pregnancy among 5 women with secondary amenorrhea. There were 3 pregnancies among 9 patients with anovulatory oligomenorrhea and 2 pregnancies in 3 patients with ovulatory oligomenorrhea. The pregnancy rate was 35%. Side effects were not observed.


[Effects of various psychotropic drugs on self-stimulation behavior in the rat]. Effects of various psychotropic drugs on tegmental and hypothalamic self-stimulation behavior in rats with chronically implanted electrodes in the brain were studied. Effects of elecrtical stimulation in several brain structures on self-stimulation behavior and influences of drugs on the stimulation effect were also investigated. The hypothalamic self-stimulation behavior was more markedly inhibited by chlorpromazine than the tegmental self-stimulation, whereas the latter was more strongly inhibited by diazepam. The effects of pentobarbital on the self-stimulation behavior were similar to those of diazepam. Methamphetamine facilitated both the tegmental and hypothalamic self-stimulation behavior. Suppressive effect of hypothalamic stimulation on the tegmental self-stimulation behavior was inhibed by chlorpromazine, while those of amygdaloid and septal stimulation were augmented. All the effects of hypothalamic, amygdaloid and septal stimulating on the tegmental self-stimulation behavior were inhibed by diazepam and pentobarbital, while these were facilitated by methamphetamine. Suppressive effects of tegmental, amygdaloid and septal stimulation on the hypothalamic self-stimulation behavior were all facilitated by both chlorpromazine and methamphetamine. The effects of amygdaloid and septal stimulation on this behavior were inhibed by diazepam and pentobarbital, while those of tegmental stimulation were enhanced. The effects of imipramine, in all experiments, were variable in each rat and not significant.


[Pharmacological studies on the cough reflex]. The artificially induced cough reflex in experimental animals should necessarily be similar to the cough which occurs naturally in humans. The curve demonstrated during a cough of each patient with respiratory disease was characteristic. Compared with the curve of a cough-like reflex (CLR) induced by DMPP, the curve of a cough induced by electrical stimulation of the tracheal mucosa was similar to that of a cough in humans. In dogs and cats anesthetized with alpha-chloralose (80 similar to 100 mg/kg i.p.), the number of CLR with DMPP was dose dependent. Tachphylaxis did not occur despite repeated administration of DMPP. DMPP (5 similar to 10 mug/kg i.v.) did not affect airway resistance. CLR with DMPP was depressed not only by morphine, codeine, oxymethebanol, picoperidamine, piclobetol and hexacol but also by hexamethonium. Thus, it appears that the mechanism of CLR with DMPP is different from that in the cough reflex. This CLR may be utilized for determining the physiology of cough reflex and the site of action of antitussives.


[Comparison of development of drug dependence in naive and drug dependence-experienced rats]. Phenobarbital, chordiazepoxide, diazepam and/or morphine were repeatedly administered to both male and female rats (N equals 10) for 4 similar to 6 weeks. The drug dose was gradully increased from 5 to 10, 20, 40, 80 and 160 mg/kg once daily (p.o) at seven day intervals. In the case of morphine, the last dose was 40 mg/kg. The drugs were constantly withdrawn for 24 hr at 8 day intervvals. None of the rats were given drugs for 16 days after administration of the last scheduled dose in order to recover their initial weight (Exp. I). Onset of dependence formation, decrease in body weight and food intake, days required to reach the maximum decrease in body weight and duration of withdrawal signs were observed throughout this experiment. The rats (drug dependence-experimented rats) who survived the first stage of this experiment were continuously subjected to re-administration by the same dosage schedule as in Exp. I (seven days of drug administration, 48hr of withdrawal). The re-administered rats showed a more rapid onset of dependence formation and a longer duration of decreas in boy weight during 16 days withdrawal than did the naive rats. It is concluded, that in addition to the decrease in body weight by withdrawal plus duration of the withdrawal signs, the onset of drug dependence formation is also a specific factor.


[Reproducibility of copper sulfate emesis by oral administration in dogs]. Twelve dogs were administered oral threshold doses of copper sulfate every week. In 66 out of 85 cases, dogs vomited. One dog vomited in 1st and 2nd tests, but did not respond in the other 6 tests. Excluding this one, the reproducibility was 82%. The following method is thus proposed for application in evaluating an antiemetic for oral copper sulfate. Small dogs, weighing 7-14 kg, are to be kept in a constant condition during the quarantine and tests and given the emetic once a week. The threshold dose is determined in three dose levels; 20, 40, 80 mg/head. The dogs with a threshold of more than 80 mg or latency of less than 5 min or of more than 45 min are to be excluded. Inhibition of emesis or a considerable prolongation of latency is the sign of an antiemetic action. A positive action of an antiemetic must be followed by another test with the threshold dose of copper sulfate alone. If the dog does not respond to the threshold dose after 2 administrations, the case must be excluded. To evaluate an antiemetic at least 5 cases are needed. Inhibition of more than 50% appears to be the positive response.


[Effects of exogenous sodium and potassium on tonus and drug-induced contractions of the pulmonary artery strip of the rabbits with special reference to their relationship to calcium]. Using the spiral strips of pulmonary artery from rabbits, effects of the modified contents of Na and K in the bath solution (BS) (Locke solution) on the tonus and the contractile responses to K, noradrenaline (NA) and Ba were investigated in relation to Ca. Removal of Na from BS by replacing NaCl with LiCl accelerated the release and influx of Ca in a short time but inhibited both 40 min later. Increase of Na in BS inhibited both the Ca influx and the Ca release from the storage site of loosely-bound Ca without affecting that of tightly-bound Ca. Evidence was obtained suggesting that Na in BS plays an important role in the uptake of intracellular Ca to the storage site. Removal of K accelerated the release and the influx of Ca in a short time but inhibited both after a long time. Data obtained suggested that Na increase or K removal inhibited the binding of NA to the adrenergic receptor. A single application of high concentrations of K (followed by washout) to the normal BS was markedly effective in restoring the preparation from its decreased contractile responses to NA or Ba following a long exposure to the Na (-), Ca (-) or the K (-), Ca (-) BS. It was suggested that this restoring effect of K is due to excitation of the cell membrane and repletion of the depleted storage site of Ca.


[Stress state caused by alteration of rhythms in environmental temperature, and the functional changes in mice and rats]. Exactly how the organic mechanism corresponds to variations of environmental temperature has not been clarified and for elucidation we reared mice and rats under the conditions of alterating rhythm in temperature (ART). Mice (or rats) were reared at a temperature of 24 degrees C and 8 degrees C (or -3 degrees C) every hour continuously from 10 a.m. to 5 p.m. and at 8 degrees C (or -3 C) from 5 p.m. to the following 10 a.m. Severe stress was evidenced in the animals and was termed specific ART-stress (SART-stress). In this type of stress, there was no apparent increase in body weight, respiration and heart rate were increased slightly and the QRS-time on the ECG was prolonged. ACh sensitivity of the isolated duodenum tested by the Magnus method in SART-stress mice was found to have declined considerably as compared to the normal. Histological observation of the SART-stress rats revealed that the wet weight of the spleen was lighter than that of the controls, while that of the lungs, heart, liver, stomach, kidneys and adrenal glands was close to that of control. Macroscopically, red brown spots of the lungs were visible. Ventricular hypertrophy, slight erosion and hyperemia were present in the interior of the stomach. On the galvanic skin response (GSR) test, electrical resistance of the skin of SART-stress rats was less than that of normal rats and the rate of increase of resistance caused by external stimulus was greater. Recovery time from change in GSR was shorter than in normal animals. Consequently, SART-stress appears to be a form of disease and sudden changes in temperature even in humans may contribute to this state.


[Effect of neurotropin on SART stress (stress caused by alteration of rhythms in environmental temperature) in mice and rats]. We reported previously that by altering the rhythm of environmental temperature (ART) the physiological mechanism was changed to an abnormal state which we termed specific ART-stress (SART-stress). In the present report, the effects of neurotropin and other tranquilizers on this SART-stress state were investigated. Neurotropin is an extract containing many conjugated polysaccarides isolated from the skin or tissues of rabbits which had been inoculated with the living cowpox virus. Neurotropin prevented the decrease in body weight of both mice and rats caused by the SART-stress. However, there was no such prevention when an extract isolated from vaccinia virus-uninjured, healthy rabbit skin was given. Chlorpromazine (Cp), reserpine (Rp), diazepam (DZ), imipramine or diphenhydramine gave no protection against the decrease in body weight. Slight increase in number and variation of respirations in SART-stress animals was to some extent prevented by neurotropin. Prolongation of the QRS interval was restored to control with the administration of the above mentioned drugs. Decrease in ACh sensitivity of the isolated duodenum tested by the Magnus-method was prevented by neurotropin. In the galvanic skin response test in rats, decrease in electric resistance on the skin, increase in reactivity and shortening of reaction time caused by SART-stress was prevented to the greatest extent by neurotropin and also to a considerable degree by parenteral administration of DZ or beta-hydroxy gamma-aminobutyric acid (GABOB), the effective results of the drugs being: Neurotropin, DZ, GABOB in that order. Cp and Rp, major tranquilizers were found to be the least effective.


[Pharmacological study of 9 alpha-fluoro-11 beta, 17, 21-trihydroxy-16 beta-methylpregna-1, 4-diene-3,20-dione-17-benzoate (betamethasone-17-benzoate, MS-1112), a local anti-inflammatory agent. (1). Its anti-inflammatory and other pharmacological properties]. The anti-inflammatory activity and the other pharmacological properties of MS-1112, a new steroid compound, were examined and compared with other glucocorticoid analogues such as hydrocortisone acetate (Hydr), betamethasone 17-valerate (Val) and dexamethasone (Dexa). Systemically administered MS-1112 and glucocorticoids had a significant effect in inhibiting rat paw edema induced by various phlogistic stimulations and increasing vascular permeabilities and granuloma formation by cotton pellet or granuloma pouch. The order of those inhibiting activity was, in general, Dexa greater than MS-1112 greater than Val greater than Hydr. Concomitantly, Xthymolysis and adrenal weight suppression and reduced rate of body weight gain after multiple systemical administration were also observed. Locally administered MS-1112 caused an inhibiting activity, without systemic side-effects. This activity was approximately 3 to 5 times more potent than that of Dexa in rat carrageenin paw edema and cotton pellet granuloma. MS-1112 was less active than Dexa in glycogen liver deposition activity and in the depression of plasma level of corticosterone but more active than Val and Hydr. In the dose administered, MS-1112 had neither androgenic and anabolic nor estrogenic and anti-estrogenic activity. From this data, it concluded that MS-1112 is a very potent agent applicable regarding permeability and retention of steroids in a local site.


[Pharmacological study on 9 alpha-fluoro-11 beta, 17,21-trihydroxy-16 beta-methyl pregna-1, 4-diene-3,20-dione-17-benzoate(betamethasone-17-benzoate, MS-1112), a local anti-inflammatory agent. (2). Its action of delayed hypersensitivity dermatitis]. The anti-inflammatory activity of MS-1112, a new steroid compound, after topical application and systemical administration was examined by means of delayed-type hypersensitivity in mice and compared with other glucocorticoid analogues, betamethasone-17-valerate (Val), dexamethasone (Dexa) and betamethasone (Beta). In the case of pretreatment and therapeutic effect, topically applied MS-1112 was most active in inhibiting the delayed-type hypersensitivity induced by picryl chloride. The order of those inhibitory activities of MS-1112 and Val was ointment greater than cream greater than gel. In contrast to MS-1112 and Val, inhibitory action of Dexa was slow in onset after the topical application. It appears that esterification of the 17-hydroxyl group increases the anti-inflammatory activity after topical application. While systemically administered, MS-1112 was more potent than Val, and similarly potent to Beta, but less active than Dexa. From the above results, it may be considered that topically applied MS-1112 is superior to other glucocorticoids in the percutaneous penetration and retention in the skin. MS-1112 is thus considered to be a very potent agent as a dermatocorticoid.


Cellular and humoral immune responses in mice. II. Effect of intraperitoneal or subcutaneous injection of carrier of anti-hapten antibody and delayed hypersensitivity responses. The subcutaneous (s.c.) injection of sheep red blood cells (SRBC) without adjuvant into mice preferentially induced delayed hypersensitivity (DH) reaction, as measured by footpad swelling, while intraperitoneal (i.p.) measured by the haemagglutinin test.  Under these conditions, the properties of the helper activities of thymus-derived (T) cells for humoral responses were examined, in association with the features of the DH response, by measuring the anti-hapten and anticarrier antibody responses after a booster injection of trinitrophenylated (TNP) SRBC and by changing the combination of doses and injections routes of the carrier and the hapten-carrier conjugates.  When mice were presensitized with i.p. injections of SRBC and boosted with i.p. injections of TNP-SRBC, the anti-TNP antibody production was maximally enhanced by presensitization with a low dose of SRBC, and gradually abolished with higher doses of SRBC for pre-sensitization.  In the latter case, anti-SRBC antibody production was increases with increasing doses of SRBC..


Activated guinea-pig C3 and the immune adherence receptor (a complement receptor) on cell membranes. By treating C3 with purified C1, C4 and C2 in the fluid phase, haemolytically inactive C3 was prepared. This was shown to bind to human erythrocytes by use of radio-labelled (Fab')2 antibody to guinea-pig C3. The activated C3 preparation inhibited immune adherence between EAC43 and human erythrocytes. These findings indicate that the activated C3 attaches to the immune adherence receptor on human erythrocytes. In addition the fluid-phase activated C3 adhered to thymus cells and sheep erythrocytes, whereas EAC43 did not. Thus the immune adherence receptor may be present on so called immune adherence-negative cells, but in insufficient concentration to form rosettes with EAC43.


Antigen-antibody complexes in or on eosinophils in nasal secretions. The presence of IgG and ragweed antigen in or on eosinophils in the nasal secretions of ragweed-sensitive patients was investigated by radioautography using specifically purified 125I-labelled antibodies to the above substances. The results indicate a significant (p smaller than 0.001) uptake of antiragweed antibody and anti-IgE antibody by eosinophils. There was no evidence of significant uptake of anti-IgG. From the above findings it was postulated that during the pollination season exposure to ragweed leads to the formation of ragweed-IgE antibody complexes. This is followed by the infiltration of eosinophils into the local tissue and the subsequent phagocytosis of these antigen-antibody complexes by the eosinophils.


Inhibition of the terminal stage of complement-mediated lysis (reactive lysis) by zinc and copper ions. The effect of various metal ions, Fe++, Fe+++, Cu++, Zn++, Co++, on the terminal stage of reactive lysis (a form of complement-mediated hemolysis in which only late-acting components of complement are required) was studied. Only Cu++ and Zn++ exhibited an inhibitory effect on the lysis of EC5678 (sheep erythrocytes reacted with C56, C7 and C8) induced by C9. The mode of action of these metal ions was further explored. Both Cu++ and Zn++ inhibited the formation of hemolytically active EC56789 from EC5678 and C9. Their effect appeared to be primarily due to the inhibition of C9 binding to EC5678 through their reversible interaction with C9. Furthermore, Cu++ is shown to inactivate irreversibly the hemolytic activity of EC5678. EC5678 pretreated with Cu++ was capable of binding C9, but the resulting EC56789 was hemolytically inactive. Besides their effect on complement, both metal ions were shown to affect directly the erythrocyte membrane, since the mechanical lysis of hemolytic intermediate cells (E, EC567, EC5678) was suppressed by Cu++ and Zn++. The lysis of EC56789, a process of internal activation, was also inhibited by both Cu++ and Zn++.


State of Trp 62 in hen egg-white lysozyme bound with N-acetylchitooligosaccharides. In order to clarify the state of Trp 62 in hen egg-white lysozyme [EC 3.2.1.17] bound with N-acetylchitooligosaccharides in relation to the ionization states of side chains present in the substrate binding site, the binding of alpha- and beta-N-acetylglucosamine GlcNAc), alpha- and beta-methyl-GlcNAc, (GlcNAc)2, and (GlcNAc)3 was studied by use of the change in the tryptophyl circular dichroic (CD) band at 295 nm at various pH values. The effects of these saccharides on the charge-transfer binding of N-1-methylnicotinamide to Trp 62 were also examined. The binding constants, estimated by use of N-1-methylnicotinamide by assuming noncompetitive binding between the saccharides and N-1-methylnicotinamide, were in good agreement with those determined from the CD change at 295 nm. The state of Trp 62 in saccharide-bound lysozyme was found to depend on the chemical structures of the saccharides and their binding orientations. The relation between the state of Trp 62 and the ionization of the catalytic Glu 35, which is far distant from Trp 62, was also confirmed for lysozyme complexes with various saccharides as well as for free lysozyme.


The binding of thyroid hormones to bovine serum albumin as measured by circular dichroism. The circular dichroism (CD) spectra of 3,5,3'-triiodothyronine-bovine serum albumin (T3-BSA) and thyroxine-bovine serum albumin (T4-BSA) complexes were measured at the various ratios of T3 or T4 to BSA (T/P) in the wavelength region of 200-350 nm. No spectral change was observed in the chromophoric wavelength region of 250-350 nm. The value of [omicron] at 319 nm of the induced Cotton effect increased with increase of the T/P ratio in both complexes, and in T4-BSA complex, it increased remarkably. The number of thyroid hormone molecules bound to one BSA molecule was estimated to be one in the case of T4-BSA complex and four to five in T3-BSA.


Studies on the primary structure of bovine high-molecular-weight kininogen. Amino acid sequence of a fragment ("histidine-rich peptide") released by plasma kallikrein. An unknown peptide fragment, which was released from bovine high-molecular-weight kininogen by bovine plasma kallikrein [EC 3.4.21.8], was isolated and its chemical structure was established. The fragment consisted of 41 amino acids with serine and arginine at the NH2- and COOH-termini, respectively. The molecular weight was calculated to be 4,584. It was very basic and contained eleven residues each of histidine and glycine and seven residues of lysine. Thus, the total number of these three amino acids accounted for about 70 percent of the total residues constituting the fragment. The amino acid sequence of the fragment, designated tentatively as "His-rich peptide," was studied by Edman degradation and standard enzymatic and chemical techniques. These data made it possible to deduce the following sequence: H-Ser-His-Gly-Leu-Gly-His-Gly-His-Gln-Lys-Gln-His-Gly-Leu-Gly-His-Gly-His-Lys-His-Gly-His-Gly-His-Gly-Lys-His-Lys-Asn-Lys-Gly-Lys-Asn-Asn-Gly-Lys-His-Tyr-Asp-Trp-Arg-OH. The fragment had an extremely interesting feature in that repeating sequences occur along the peptide chain. The repeats were of the type His-Gly-X or Gly-His-X and this sequence appeared six or seven times up to 26 residues from the N-terminal end. Moreover, three tetrapeptide sequences of Gly-His-Gly-His and two heptapeptide sequence consisting of His-Gly-Leu-Gly-His-Gly-His were found in the N-terminal portion. It should be noted that plasma kallikrein liberates such a histidine-rich peptide from the kininogen in addition to a physiologically active peptide, bradykinin. The location of the "His-rich peptide" fragment in the percursor protein is also discussed.


Principle of operation of the thermal energy analyzer for the trace analysis of volatile and non-volatile N-nitroso compounds. The theoretical basis for the thermal energy analyzer is discussed. Using the principles outlined, the feasibility of selectivity detecting mug/kg levels of volatile and non-volatile N-nitroso compounds is established.


A rapid modified gas chromatographic assay for esterase activity. A rapid modified procedure for the gas chromatogrpahic determination of esterase activity was studied. Aliphatic esters such as ethyl n-butyrate, n-propul n-butyrate, n-butyl n-butyrate and n-amyl n-butyrate were used as substrates and acetone was chosen as the most suitable solvent for dissolving the substrates in order to avoid alcoholysis. The enzyme reaction was started in a mixture of 0.03 M phosphate buffer, pH 7.90, containing an adequals, an aliquot of the reaction solutionwas injected directly on to a gas chromatograph and the alcohols produced were separated.


Examination of the diphenylpropanoids of nutmeg as their trimethylsilyl, triethylsilyl and tri-n-propylsilyl derivatives using combined gas chromatography and mass spectrometry. Ethyl acetate extracts of thirteen samples of nutmeg and one sample of mace were examined as trimethylsilyl and tri-n-propylsilyl derivatives by gas chromatography and mass spectrometry. Eleven compounds derived from two types of diphenylpropanoid were identified; the relative proportions and quantities of these compounds varied considerably in the different samples. Single ion chromatograms were used to obtain the relative concentration of one series of compounds. Preparation of the triethylsilyl and tri-n-propylsilyl derivatives resulted in greatly increased separation of the diphenylpropanoids over trimethylsilyl derivatives, which tended to produce a group of compounds with many unresolved peaks.


Pubertal sleep-wake patterns of episodic LH, FSH and testosterone release in twin boys. On 2 consecutive nights, plasma LH, FSH and testosterone (T) were measured every20 min for 12 h during evening wakefulness and polygraphic sleep in 5 pairs of male monozygotic twins in pubertal stages 1-4, and in a male dizygotic also studied in 3 twins. During sleep, significant enhancement of episodic LH release was seen on 16 of 18 nights on the stage 1-4 twins. During wakefulness, minimal episodic LH release was observed in the stage 1-3 twins, which then gradually increased in the more mature twins, until finally the significant sleep-wake difference in mean LH was lost in the stage 5 male. Testosterone also rose significantly in sleep on 19 of 20 study nights in the stage 1-5 twins. In the early pubertal twins this nocturnal rise in T was small, but in the midpubertal pairs it was profound, as peaks in T occurred which lay in the normal range for adult males. In these less mature twins the majority of the episodic secretion of T also was limited to sleep. In wakefulness, the T levels gradually increased across puberty until, in the stage 5 twin, wakeful peaks in T finally reached the adult male range. In the midpubertal twins, a close temporal relationship was seen between initiation of sleep-enhanced LH release and the subsequent initial rise in T (mean lag time 29.1 min). In the stage 5 twin, this episodic LH-T relationship persisted into wakefulness where the largest increments in T were seen just prior to sleep onset. Evidence of sleep-enhanced FSH release was more equivocal, and was limited mainly to pubertal stage 1 and 3 pairs. Similarities in hormonal patterns were seen within the monozygotic twin pairs and probably contributed to the parallel progress in puberty of the pair. Thus, sleep-wake rhythmicity in release of gonadotropins, particularly LH and thereby of testosterone, was seen to evolve transiently in twin boys across puberty. The existence of such rhythmicity suggests that a fundamental, sleep-entrained CNS mechanism plays an important, if not a dominant, role in sexual maturation in boys.


Inhibition of human platelet aggregation by monovalent antifibrinogen antibody fragments. Monovalent goat antibody fragments (Fab) that were monospecific for human fibrinogen were isolated by affinity chromatography on fibrinogen-Sepharose and used as a direct probe for the involvement of fibrinogen in platelet aggregation and the release reaction. The antifibrinogen Fab inhibited aggregation of washed human platelets induced by thrombin (0.1-10 U/ml) by 50-95%, but had no effect on (14-C)-serotinin release and only a slight inhibitory effect on 125-I-thrombin binding to platelets. Inhibition of aggregation was not observed with nonimmune goat Fab or rabbit antihuman albumie bound tightly at saturation to surface fibrinogen molecules. After washing the platelets once to remove unbound Fab, aggregation by subsequently added thrombin was no longer inhibited. The antifibrinogen Fab inhibited the clotting of fibrinogen by thrombin but did not effect the rate of fibrinopeptide A release, indicating that the Fab inhibits clotting by interfering with the polymerization of fibrin monomers. Our experiments suggest that fibrinogen released from platelets is directly involved in thrombin-induced aggregation of washed platelets, perhaps through polymerization of fibrin monomers generated by proteolytic cleavage of released fibrinogen.


The structure of the inferior lobe of the teleost hypothalamus. Electron microscopic and Golgi studies on the inferior lobes of sunfish and goldfish are described. The inferior lobe consists primarily of a nucleus ventricularis of densely packed cells surrounding the lateral recess of the third ventricle, and a peripherally situated nucleus diffusus consisting mostly of scattered neurons. A cell-sparse zone of dense neuropil is located between the two cellular areas. Neurons of both nuclei have spiny dendrites and axons which originate from basal dendrites. In some cases axons are found to send a collateral into the cell-sparse zone. Neurons of the nucleus diffusus possess collaterals that extend a considerable distance within the nucleus itself. The ultrastructure of cells of both nuclei reveals cytoplasmic organelles typical of most neurons. Synapses containing dense-cored and clear vesicles are present on the spines and shafts of the dendrites of both neuronal types. In only rare cases synapses were observed on the soma of neurons of the nucleus ventricularis. Possible anatomical substrates involved in the control of feeding and aggression in teleosts are considered in light of the present findings. Morphological similarities of the inferior lobes and related areas in various fishes and amphibians are discussed and their possible significance for the understanding of the evolution of hypothalamic mechanisms is considered.


Hepatitis B surface antigen in saliva of HB-SAg carriers. Two hundred and twelve saliva specimens of HB-SAg carriers were tested for HB-SAg by crossover immunoelectrophoresis (CIEP) and radioimmunoassay (RIA) after concentration, between April 1971 and September 1973. They were divided into 3 groups (A, B, and C). CIEP detected HG-SAg in 6 per cent in Group A (66 specimens) and in 12 percent in Group B (87 specimens), and in 78 per cent in Group C. Different methods of preparation and concentration of saliva specimens were discussed. The positive results by RIA were confirmed by a blocking test with rabbit HB-SAg antiserum. Intermittent presence of HB-SAg in repeated saliva specimens from individual carriers was found. Infected saliva of HB-SAg carriers may be of importance in transmission of hepatitis B infection.


Effects of ovarian steroid hormones on esterases in the rat endometrium. A technique in which uterine luminal epithelium is separated from the remainder of the endometrium by rapidly vibrating everted uterine cornua in a 3-5 mM solution of EDTA has been developed. Examination of the hormonal sensitivities and physiological roles of the tissue components of the endometrium is thus facilitated. Biochemical and histochemical studies of epithelial, stromal and endometrial esterases have shown that the rate of hydrolysis of alpha-naphthyl acetate is significantly higher in epithelial and endometrial tissue extracts during pro-oestrus than at any other stage of the oestrous cycle. In ovariectomized animals, oestradiol-17 beta caused a 60% increase in the rate of esterase activity over that of control animals, whereas medroxyprogesterone acetate had no effect. These findings suggest that the variations in the levels of neutral lipids in the uterine luminal epithelium of non-pregnant mature female rats result from the periodic stimulation of the epithelial esterases by the cyclically increased levels of plasma oestrogens.


Influence of anaesthetics, mating, premature delivery, the time of day of blood sampling, and suckling on plasma levels of luteinizing hormone in hamsters. The influence of several factors on immunoreactive LH levels in peripheral plasma of female hamsters was investigated. The LH values for animals anaesthetized with either sodium pentobarbitone or chloral hydrate were significantly lower (P smaller than 0-01) than those values obtained for hamsters sampled under ether anaesthesia. Mating on the night of pro-oestrus resulted in increased LH levels (P smaller 0-01) the following morning as compared with the levels in control females not caged with males. The LH levels were relatively low (13 plus or minus 2 (S.E.M.) ng/ml) on the afternoon (16.00-18.00 h) of day 15 of pregnancy and also on the morning (09.00-10.00 h) of day 16 before (7 plus or minus 1 ng/ml) and after parturition (4 plus or minus 0-2 ng/ml), but rose severalfold (561 plus or minus 201 ng/ml) on the afternoon post partum. When premature delivery was induced on day 15 of pregnancy by administration of prostaglandin E2 the LH levels did not rise on the afternoon of this day (29 plus or minus 11 ng/ml), but did so on the following afternoon (213 plus or minus 115 ng/ml), at the same interval post coitum as in normal delivery. Marked increases in the concentration of LH in plasma were observed on the afternoon of each of days 0-6 post partum. The levels were comparable to those found on the afternoon of pro-oestrus in cyclic hamsters and could be blocked by administration of sodium phenobarbitone at 13.00 h. Similar afternoon surges of LH were not observed during dioestrus. In the morning, the LH levels were significantly higher (P smaller 0-001) in the non-suckled dams (95 plus or minus 4 ng/ml) than those in mothers allowed to nurse their young (13 plus or minus 4 ng/ml). In contrast, the levels in the afternoon were unaffected by the suckling stimulus (suckled, 495 plus or minus 76 v. non-suckled, 744 plus or minus 165 ng/ml; P greater than 0-05).


The effect of daylength and level of feeding on serum prolactin in growing lambs. Twenty-four castrated male lambs initially maintained on a photoperiod of 12 h light:12 h dark were allocated to a factorial experiment with two daylengths (8 h L: 16 h D or 16 h L: 8 h D) and two levels of feeding (restricted or ad libitum). Blood samples were taken every 4 h for 24 h during the introductory period and after 24, 51 and 79 days of treatment. There were highly significant positive effects of daylength and level of feeding on serum prolactin: mean concentrations increased from a mean of 38 plus or minus 1 ng/ml during the introductory period until at day 79 they were: 8L: 16D (restricted diet), 81 ng/ml; (food ad libitum), 167 ng/ml; 16L:8D (restricted diet), 262 ng/ml; (food ad libitum), 262 ng/ml (S.E. of treatment mean plus or minus 4). Long daylength and feeding ad libitum also significantly increased growth rate.


Effect of supplements of zinc salts on the healing of incised wounds in the rat and guinea pig. Zinc deficiency affects healing as shown by measurement of two parameters of wound healing in the rat, namely the rate of healing of an excised, granulating wound and the development of tensile strength in an incised wound. The rate of healing of granulating wounds in two species of animals, the rat and the guinea pig, is not affected by the addition of supplements of zinc salts to the normal diet (J. Nutr. 105, 815-821). In this study, the development of tensile strength in incised wounds in rats and guinea pigs was measured at 7 and 14 days after wounding in animals given supplements of zinc salts by either the oral or parenteral route. No difference in tensile strength was observed at these times in the wounds of either rats or guinea pigs given zinc supplements.


Lipid peroxidation and glutathione peroxidase activity in the liver of cholesterol-fed rats. The effects of cholesterol feeding on lipid peroxidation and on glutathione peroxidase activity in the liver of rats were examined. Cholesterol (1 or 1.5%) was added to two basal diets containing either 10% soy oil (diet 1) or 15% soy oil plus 100 IU of alpha-tocopherol/kg of diet (diet 2). The rate of lipid peroxidation in the liver was determined in vitro by the thiobarbituric acid test and by conjugated diene measurement (234 nm). Cholesterol feeding elevated the rate of lipid peroxidation in the liver of rats fed diet 1 or 2. The rate of lipid peroxidation in rats fed diet 2 increased with duration of feeding suggesting an increased need for antioxidant as the levels of cholesterol or other lipids in the liver elevated. Cholesterol feeding also decreased the activity of liver glutathione peroxidase and pentose phosphate pathway dehydrogenases. Glutathione peroxidase activities increased with the duration of feeding in rats fed basal diet 2, but they remained unchanged in rats fed the same diet supplemented with cholesterol. The study suggests that cholesterol feeding in rats can increase the susceptibility of the tissue to lipid peroxidation and also can lead to a depression in the activity of glutathione peroxidase.


Risk of infection associated with umbilical vein catheterization. A prospective study in 75 newborn infants. To determine the risk of infection created by catheterization of the umbilical vein per se, 75 jaundiced, but otherwise healthy, newborn infants subjected to exchange transfusion were studied. Twenty-three were given antibiotics because of premature rupture of membranes. Fifty-three percent of the umbilici were contaminated before the insertion of the catheter, even after a very careful cleansing procedure. Sixty-two percent of the catheters were colonized upon removal. Bacteria were isolated from 44.9% of blood specimens drawn via the catheter at the onset of ET, but only in 14% of blood specimens drawn in the same way at the end of the procedure. Seven newborn infants (10%) were found to be bacteremic 4-6 after ET; four of these infants were not treated and were able to eliminate the bacteremia. Systemic antibiotic therapy did not reduce the overall prevalence of colonization of cord and catheter of positive blood cultures pre- and postexchanges transfusion or the rate of bacteremia.


Aerobic and anaerobic carbohydrate metabolism and egg production of Schistosoma mansoni in vitro. When S. mansoni adults were cultured in vitro for 12 days in a diphasic medium, their gross morphology, motor activity, frequency of sexual pairings, rates of glucose utilization and of lactic acid production were the same in the presence (90% N2/5% O2/5% CO2) or absence (95% N2/5% CO2) of oxygen. Therefore, no Pasteur effect, nor any reduction in lactic acid formation, was demonstrable under aerobic conditions. While aerobic conditions did not affect the rate of glycolysis, they had a marked effect on egg production. In the presence of oxygen, the rate of egg-laying reached a maximum between days 4 and 6. The average number of viable eggs produced per worm pair during this period was 118 (Sx equals 2.2), which is within the overall range (68 to 248) recorded by others for this same strain in vivo. Conversely, under anaerobic conditions in vitro, virtually no eggs were laid. It remains to be determined whether oxidative metabolism actually is required for energy to produce eggs, or whether some reaction yielding no ATP is essential for completion of their developmental process, such as tanning of the eggshall brought about by the oxidation of some phenolic compounds.


Potential anticancer agents II: antitumor and cytotoxic lignans from Linum album (Linaceae). A phytochemical study of Linum album (Linaceae), guided by bioassay with the 9KB cell culture, resulted in the isolation of podophyllotoxin and a new lignan, 3'-demethylpodophyllotoxin; alpha-and beta-peltatins were identified by comparative TLC.


Decreased 6-mercaptopurine retention by two resistant variants of mouse neuroblastoma with normal hypoxanthine-guanine-phospho-ribosyltransferase activities. In an effort to propose more effective chemotherapeutic regimens for the treatment of neuroblastoma, we have characterized mouse, neuroblastoma variants whose growth in tisue culture are resistant to antimetabolites. We report the partial characterization of two lines resistant to 6-mercaptopurine (6-MP). Concentrations of drug required to inhibit their growth rates 50% are 110- and 575-fold higher, respectively, than that inhibiting the sensitive parental clone. Unlike most 6-MP-resistant cell lines described previously, both neuroblastoma populations display normal activities of hypoxanthine-guanine phosphoribosyltransferase but greatly reduced accumulation of 14-C-labeled 6-MP. Drug accumulation was inhibited by adenine, blocked by dinitrophenol but not ouabain and strongly temperature dependent suggesting a need for cytoplasmic phosphoribosylation. Possible mechanisms for this reduction in 6-MP retention are discussed. Importantly, eight clones isolated in 6-MP-free media from the 110-fold resistant population of cells demonstrated quantitatively identical growth inhibition at all drug concentrations tested suggesting that the original 110-fold resistant neuroblastoma population was homogenous with respect to its mechanisms of resistance.


Effects of cholinergic and anticholinergic drugs and a partial cholinergic agonist on the development and expression of physical dependence on morphine in rat. Cholinergic and anticholinergic agents were tested against the induction and expression of morphine dependence in rats. Mecamylamine hydrochloride, administered during abstinence, exacerbated the syndrome at the time of peak severity (20-40 hours after with-holding morphine) but accelerated the subsequent decline in severity of withdrawal. Atropine sulfate acted similarly but none of its effects proved statistically significant. When administered together, these drugs were not additive but were antagonistic in their central actions at the time of peak withdrawal. Atropine methylnitrate reduced withdrawal severity throughout the observation period mainly by antagonism of the peripheral expression of many of the parasympathetic signs. Eserine sulfate administered during the period of habituation or of abstinence had little effect on the severity of the total withdrawal syndrome but decreased weight loss. Choline chloride effectively reduced overall withdrawal severity and weight loss throughout the entire course of withdrawal monitored for up to 100 hours after withholding morphine. This occurred whether choline was administered during morphine habituation or after withholding the morphine. These results support the view that a derangement of cholinergic function is a factor in the morphine abstinence syndrome; evidence is yet lacking to decide whether this is the primary derangement of neurotransmitter function. The effectiveness of choline may be explained by its action as a partial agonist to normalize cholinergic function at all stages of derangement. The reduction in withdrawal severity by treatment with choline during the habituation to morphine indicates an inhibition of the development of dependence rather than mere suppression of the signs during withdrawal.


Action potential and non-linear current-voltage relation in starfish oocytes. 1. The electrical properties of the oocyte membrane of the starfish, Asterina pectinifera, were investigated using an intracellular microelectrode. 2. The resting potential in artificial sea-water ranged from -70 to -80 mV. 3. The starfish oocyte membrane was capable of generating an action potential as a result of permeability increases to both Ca and Na ions.4. The Ca component of the action potential was reversibly suppressed by Co or Mg ions, while the Na component was not affected by tetrodotoxin 5 times 10-minus 6 g/ml. 5. The steady-state relation of voltage vs. current was not linear but S-shaped. The curve wascomposed of inward-going rectification at the membrane potential more negative than -65 mV, outward-going rectification at the potential more positive than OmV and the transitional region between them. These findings are compared with those obtained in the mature egg of the tunicate.


Potassium rectifications  of the starfish oocyte membrane and their changes during oocyte maturation. 1. The current-voltage relations of the oocyte membrane of the starfish, Asterina pectinifera, and their changes during maturation were investigated using current-clamp techniques. 2. The resting potential of the oocyte membrane in sea water was found to be determined by the diffusion potential of K ions. 3. In the absence of Na and Ca inward currents the steady-state current-voltage relation of the oocyte membrane had inward-going K rectification at membrane potentials more negative than -65 mV and outward-going K rectification at potentials more positive than -30 mV, forming a S-shaped I-V curve. 4. A negative resistance region of the steady-state I-V curve was revealed with voltage-clamp technique in the potential range between -65 and -30 mV. 5. Transient K activation occurred when the membrane was brought from a resting potential of about -75 mV to potentials more positive than -20 mV, and this was immediately followed by K inactivation. Accordingly, the steady-state I-V relation showed only slight outward-going rectification. 6. At the beginning of meiosis, which is signalled by break-down of the nucleus, the limiting slope conductance in the inward rectifying region of the I-V curve decreased sevenfold. The cell membrane lost its selective permeability to K ions and was depolarized from -70 to between -20 and OmV in standard artificial sea-water. The depolarized resting potential was partly due to the relative increase in Na permeability. K conductance began to increase again within 30 min after breakdown of the nucleus. The resting potential became gradually larger and eventually attained -70 mV in the mature egg. 7. In the mature egg, K activation upon depolarization was no longer followed by inactivation. Accordingly, the slope conductance in the outward rectifying region of the I-V curve increased. 8. The action potential was augmented at the stage of nuclear breakdown. Thereafter the maximum rate of rise decreased and the duration of the action potential shortened. These changes were caused primarily by changes in K conductance during maturation. 9. Fertilization of the egg during the maturation process did not affect the changes in the I-V relation described above, except for a transient change of the membrane permeability upon fertilization.


Relationships between the biological and physicochemical properties of series of compounds. An equilibrium model is described for relationships between the biological activity and the physicochemical properties of compounds in a congeneric series. An equation derived from the model is of the form, log C = log (a + 10-alpha) = b, where C is the total molar concentration of drug to produce a standard response, alpha ia a parameter of the form log K where K is an equilibrium constant, and a and b are coefficients. This equation provides a plausible fit to some nonlinear observations which have been studied.


Synthesis and antifolate activity of new diaminopyrimidines and diaminopurines in enzymatic and cellular systems. The following new 2,4-diamino-6-methylpyrimidines, 5-cyclohexylmethyl, 5-cyclohexylethyl,and 5-(2-naphthyl), as well as 2,6-diaminopurines, 8-adamantyl and8-adamantylmethyl, were synthesized as potential antifolates. Tese, as well as three known compounds, 2,4-diamino-5-cyclohexyl-6-methylpyrimidine, 2,4-diamino-5-(1-naphthyl) -6- methylpyrimidine, and 2,6-diaminopurine, were compared with respect to the inhibition of growth of mammalian cells in culture (TA 3) and with respect to the inhibition of partially purified dihydrofolate reductase. All of the pyrimidines except for the 5-(1maphthyl) derivative were competitive inhivitors of dihydrofolate reductase, with K values ranging from 0.07 to 0.04 pM. They were 2-5 times better as inhibitors of the isolated dihydrofolate reductase than of the cell growth. 2,4-Diamino-5-(1-naphthyl)-6-methylpyrimidine was a noncomptive inhivitor of the enzyme with a Kvalue of 56 pM. This compound was more potent in inhibiting cell growth than the isolated enzyme. indicating that its biological activity was not related to the inhibition of dihydrofolate reductse. All of the purine derivatives were poor growth inhibitors and although some of them inhibited isolated dihydrofolate reductase, their mode of action in cellular system did not seem to concern folate metabolism, as judged by the inability of hypoxanthine, and glycine to provide protection. The implication of these findings as to the structural requirements for inhibition of dihydrofolate reductase is discussed. The implication of these findings as to the  structural requirements for inhibition of dihydrofolate reductase is discussed. The pitfalls of the determinition of ID-50 values instead of a complete kinetic analysis in structure-activity studies are emphasized.


Base-catalyzed and cholinesterase-catalyzed hydrolysis of acetylcholine and optically active analogs. The base- and cholinestrase-catalyzed hydrolyses of the following optically active analogs of acetylcholine were studied: 3 (a)-trimethylammonium-2(a)-acetoxy-trans-decalin iodide, threo- and erythro-alpha, beta-dimethylacetylcholine iodide, alpha-methylacetylcholine, and beta-methylacetylcholine. Evidence that the optimum dihedral +N-C-C-O angle in the transition state for acetylcholinesterase hydrolysis of acetylcholine analogs is positive and anticlinal is given. The data obtained suggest that acetylcholine undergoes a geometrically flexible mode of attachment to the enzyme.


Perceived sources of stress among first-year medical students. A survey of first-year medical students at the University of Toronto shows that academic factors are viewed as the most stressful and social factors as the least stressful. Among anticipated sources of stress those dealing with perceived failure are highly stressful, and those dealing with sexual aspects of the doctor-patient relationship are much less so. Factor analysis reveals that sources of stress are multidimensional rather than unidimensional. Particular socio-demographic antecedent variables are much more highly correlated with stress than are others. In particular, it is found that students in subgroups differing from "mainstream" students feel more stress than their mainstream counterparts. Feelings of stress are also found to be positively related to possible consequences of stress, such as frequency of thinking about dropping out and to number of days off school because of illness.


In vitro suppression of immunocompetent cells by lymphomas from aging mice. We studied the effects of lymphomas in the spleens of aging (C57BL/6 X BALB/c)F1 (B6CF1) mice on the immunologic responsiveness of normal spleen cell cultures to sheep erythrocytes. The cultures were assayed for antibody-forming cells (AFC). Cultures of 20 times 10-6 normal spleen cells produced hundreds of AFC, whereas similar cultures of lymphomatous spleen cells produced less than ten AFC. Normal spleen cells were immunosuppressed in cultures with as few as 10-4 lymphomatous spleen cells. Our results demonstrated that lymphoma cells inhibited the response of normal spleen cells to antigenic stimulation in vitro. The age response for male mice gave a biphasic decline with a steeper second phase. This second decline of immune responsiveness started to appear as the incidence of lymphomas rose. This suggested that the immunosuppressive activity of lymphoma cells was causally related to the age-dependent decline of immune responsiveness in B6CF1 mice.


Transplacental induction of primary renal tumors in rabbits treated with 1-ethyl-1-nitrosourea. Pregnant rabbits of the two partially inbred strains III and WH were given ip injections of a single dose of 1-ethyl-1-nitrosourea (ENU) (60 mg/kg) in trioctanoin on day 18 of gestation. Controls were treated on the same day with solvent alone. Fourteen of 15 strain III progeny that survived more than 8 weeks developed primary renal tumors at a mean age of 3.3 months. Five other treated strain III progeny died at an early age due to other causes. In contrast, only 3 of 7 strain WH offspring surviving more than 8 weeks developed renal tumors; they had about the same latency period (3.9 months). In each strain, either renal tubular cystadenomas or mixed nephroblastomas appeared to develop within small renal cortical cysts. In strain III, the presence of these cysts may have been due to a high frequency of a recessive gene (rc) for renal cysts, but in strain WH they were induced by ENU. The differential strain incidence suggests that susceptibility to renal tumor inducibility by ENU is increased by the presence of the rc/rc genotype for cyst formation.


EEG evaluation of humaneness of asphyxia and decapitation euthanasia of the laboratory rat. The relative humaneness of asphyxia and decapitation was objectively evaluated in rats by EEG monitoring. EEG activation (low voltage, fast activity) was considered to indicate discomfort, pain, and affective responses to euthansia. Such activation was present 37.3 plus or minus 7.5 sec after asphyxia and 13.6 plus or minus 4,6 sec after decapitation. Decapitation was also characterized by an immediate large, and relatively long-lasting, ultra-slow voltage, detected by non-polarizable scalp electrodes. Isoelectric activity (death) occurred 69.4 plus or minus 9.9 sec after onset of asphyxia and 27.2 plus or minus 4.4 sec after decapitation.