Prostaglandin E2 tablets compared with intravenous oxytocin in induction of labour. Stimulation of uterine activity after amniotomy has been carried out with prostaglandin E2 (PGE2) tablets in two dosage regimens and with intravenous oxytocin. Oxytocin stimulation was the most successful. The difference in success rate was most marked in nulliparous patients and those with low Bishop score.


Delay in diagnosis of optic nerve and chiasmal compression presenting with unilateral failing vision. Out of 29 patients who presented with failing vision in one eye due to optic nerve or chiasmal compression, compression was initially diagnosed in only five. The errors in diagnosis and lack of ophthalmological follow-up led to delays of up to many years with serious deterioration in acuity before referral for intracranial investigation. The chief causes of error were lack of charting of the visual fields, too ready acceptance of the diagnosis of the neuritis in the absence of essential features, and the infrequent use of skull radiographs.


Thyroid function after subtotal thyroidectomy for hyperthyroidism. Among 76 patients who had had a subtotal thyroidectomy for hyperthyroidism from one to seven years previously recurrent hyperthyroidism was found in three and hypothyroidism in 13. The remaining 60 subjects were clinically euthyroid but a raised level of serum thyroid-stimulating hormone (TSH; greater than 5-0 mu U/ml) was found in 39. Analysis of the data showed that their serum thyroxine was significantly lower than in the subjects with a normal TSH. The serum triiodothyronine (T-3) was similar in both groups. It is concluded that subjects with a raised TSH remain clinically euthyroid by maintaining a normal serum T-3 concentration. There was no evidence of any long-term progressive deterioration of thyroid function after subtotal thyroidectomy.


Use of charcoal haemoperfusion in the management of severely poisoned patients. The clinical use of uncoated charcoal haemoperfusion systems, despite their efficacy, has hitherto been prevented by the occurrence of a number of adverse effects including charcoal embolism and marked thrombocytopenia. Charcoal coated with a synthetic hydrogel overcomes many of the disadvantages associated with the use of uncoated material in that there is a much reduced thrombocytopenia and no evidence of charcoal embolism. Six patients, severely poisoned as a result of overdoses of either a barbiturate or glutethimide, were haemoperfused using such a system. Four made complete recoveries, and the two patients who died had both suffered cardiorespiratory arrests before perfusion. In contrast to haemodialysis charcoal haemoperfusion is simple to initiate, less expensive in terms of manpower and equipment, and gives superior clearance data for all barbiturates and glutethimide. We believe that this technique may have a significant role to play in the management of the severely poisoned patient.


Localization of the cockroach optic lobe circadian pacemaker with microlesions. Electrolytic microlesions were made at various sites in optic lobes of cockroaches Leucophaea maderae, and animals were assayed for circadian locomotor rhythms. Lesions placed in the lamina or medulla disrupted rhythmicity in only a few animals (3 out of 55). Lesions placed in or near the lobula produced a greater fraction of arrhythmic roaches (18 out of 45). Over half of these lesions either included or were centered in the cell body regions bordering the second optic chiasm and the lobula. The results suggest that the cell bodies and not the neuropile areas of the lobe are the crucial elements of the clock driving the cockroach's circadian activity rhythm.


The morphometry of the branching pattern in dendrites of the visual cortex pyramidal cells. An analysis has been made of the three-dimensional branching structure for the basal and apical dendrites of cortical neurons in an adult rabbit. The real branching angles of basal dendrites and apical oblique branches are in the same range, but differ from those of the apical main shaft. Therefore, several different parts of the apical dendrite have to be distinguished on anatomical grounds, coincident with the presynaptic areas distinguished in the literature. The bifurcations of basal dendrites are essentially symmetrical. The mode of outgrowth, however, is non-symmetrical. Redirection of dendrites will, therefore, occur. This redirection is often not complete, so that a large variability of branching angles results. The possible significance of the observed symmetry is discussed.


Synthesis and some biological properties of 1-deamino-4-glu-oxytocin (1-beta-mercaptopropionic acid-4-glutamic acid-oxytocin) and its use in preparing a hormone-agarose complex. 1-Deamino-4-glu-oxytocin (1-beta-mercaptopropionic acid-4-glutamic acid - oxytocin) was synthesized by sequential reduction by sodium in liquid ammonia and oxidation by hydrogen peroxide of the octapeptide derivative, S-benzyl-beta-mercaptopropionyl-tyrosyl-isoleucyl-gamma-O-benzyl-glutamyl-asparaginyl-S-benzyl-cysteinyl-prolyl-leucyl-glycinamide. The oxidation analogue was isolated and purified by partition chromatography in two different solvent systems followed by exclusion chromatography on Sephadex G-25. It was found to possess approximately 13 I.U. of uterotonic activity, 34 I.U. of milk ejection activity, and 83 I.U. of milk ejection-like activity per milligram, measured on an isolated strip of lactating mouse mammary gland. 1-Deamino-4-Glu-oxytocin was coupled to AH-Sepharose 4B by the way of the free gamma-carboxyl group of its residue of glutamic acid. The water soluble 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride caused the coupling with approximately 70% effectiveness. The resultant peptide-agarose complex had low biological potency in the assay of milk ejection-like activity.


Insulin control of hepatic glucose production. Insluin injected intravenously caused a rapid, marked decrease in hepatic glucose secretion in the rabbit, as determined by an isotope-dilution procedure. This was associated with a decrease in the concentrations of gluconeogenic intermediates from phosphoenolpyruvate to triose phosphates, inclusive, compatible with inhibition of gluconeogenesis at phosphoenolpyruvate carboxykinase. The concentration of glucose 6-phosphate was unaltered but that of hepatic glucose was reduced. The specific activities of the hexose phosphates, relative to that of liver glucose, were the same in control and insulin-treated animals. These observations can be explained by a decrease in the activity of glucose-6-phosphatase. It is concluded that this enzyme is a control point for hepatic glucose production and is inhibited by insulin. In the rat, insulin produced a rapid fall in blood sugar. The hepatic glucose output remained normal despite a fall in hepatic glucose 6-phosphate concentration during the initial period of insulin action. This suggests that glucose-6-phosphate returned to normal with no change in the rate of glucose production. The data suggest that in the rat, insulin produces a transient increase in glucose-6-phosphatase activity.


3-Methyl-2-butenal: an enzymatic degradation product of the cytokinin, N-6-(delta-2 isopentenyl)adenine. An enzyme preparation from immature corn kernels catalyzed cleavage of N-6-(delta-2-isopentenyl)adenine to give the aldehyde, 3-methyl-2-butenal, as the major side-chain derived product. This product, in the form of the semicarbazone, was identical with an authentic product by several criteria: chromatographic behavior, mass and ultraviolet spectra.


Metabolism distribution, and disappearance of injected beta-phenylethylamine in the rat. In the absence of a monoamine oxidase inhibitor, the bulk of intravenously injected radioactively labelled beta-phenylethylamine was oxidized to phenylacetic acid. In the presence of pargyline, most of the label in tissues remained as unchanged phenylethylamine; small amounts of labelled phenylethanolamine, tyramine and octopamine were also identified. After intravenous injection of [14-C]phenylalanine, only very small amounts of [14-C] phenylethylamine could be located in urine and faeces. Beta-Phenylethylamine became concentrated in all tissues, including brain, following intravenous introduction both in the presence and absence of pargyline. Its clearance from these tissues and from brain regions was very fast.


Changes in metabolism of ethanolamine and its derivatives in liver during fasting. The time course of incorporation of radioactivity into liver phosphatidylethanolamine and its precursors was studied following intraportal injection of [1,2-14C]ethanolamine into rats that had been fasted for 24 h and into nonfasted rats. Marked diminution in the labelling of liver phosphatidylethanolamine and CDP-ethanolamine was present in the fasted groups while the radioactivity contained in the ethanolamine and phosphorylethanolamine pools of these livers was increased. The livers of the fasted rats contained significantly higher levels of ethanolamine, phosphorylethanolamine and CDP-ethanolamine while choline and phosphorylcholine were decreased. These changes are identical to those which we have described earlier in the livers of nonfasted rats fed a choline deficient diet. In a further experiment, the duration of the fasting period necessary to bring about these changes was studied. Incorporation of radioactivity into liver phosphatidylethanolamine at 2.5 min following intraportal injection of [1,2-14]ethanolamine declined progressively up to 7.5 h of fasting and did not decrease further after that time. There was a concomitant decrease in labelling of CDP-ethanolamine with retention of radioactivity in ethanolamine and phosphorylethanolamine. These results show that the liver metabolism of ethanolamine is altered quite early in the fasting period and suggest that it may be related in some way to the metabolic response to fasting.


Identification and distribution of m-tyramine in the rat. A procedure for the quantitative evaluation of m-tyramine in mammalian tissues is described. It involves isolation of the amine by ion-exchange chromatography, followed by conversion to the dansyl derivative, chromatographic separation, and quantitation by the mass spectrometric integrated ion current technique using an isotopically labelled internal standard. The concentrations of m-tyramine in some tissues of male Wistar rats were (mean plus or minus S.D., nanograms per gram): brain 0.32 plus or minus 0.03, heart 0.44 plus or minus 0.13, kidney 12.6 plus or minus 3.4, liver 0.27 plus or minus 0.04, lung 0.33 plus or minus 0.11, spleen 0.25 plus or minus 0.07, and blood 0.15 plus or minus 0.04.


RNA polymerase stimulation: effect of aldosterone and other adrenocorticoids on RNA turnover in rat kidney. The results of steroid hormone stimulation of aggregate RNA polymerase ?activity in kidney can be interpreted in terms of either template or enzyme alterations. In order to discover the effect of aldosterone on enzyme or DNA template, the RNA polymerases were purified from kidney nuclei of normal, adrenalectomized or adrenalectomized plus aldosterone treated rats and the activity was determined using different sources of DNA. It was found that the DNA from aldosterone treated rats was transcribed more efficiently than from other sources. Aldosterone does not seem to have a direct effect on the RNA polymerase. Fractionation of ([14-C]aldosterone injected) kidney chromatin revealed the presence of radioactivity in the non-histone acidic proteins and DNA, suggesting the possible binding of aldosterone or aldosterone-receptor complex to these chromatin fractions which may result in enhanced template activity. Turnover of RNA was also studied in various subcellular fractions: nuclei, mitochondria, rough endoplasmic reticulum, smooth endoplasmic reticulum, polysomes, ribosomes and sRNA, in normal, adrenalectomized, and adrenalectomized plus adrenocortical hormone treated rat kidney, by following the loss of radioactivity after a single injection of [14-C]-orotic acid. Daily administration of aldosterone or deoxycorticosterone reversed the effects of adrenalectomy. Daily administration of corticosterone was without effect.


Isolation and tissue specificity of chromatin-associated proteins in Vicia faba. A method is described that permits extraction of one class of non-histones in 8 M urea minus 0.14 M mercaptoethanol prior to acid extraction of histones and a second class in 0.05 M Tris -1% sodium dodecyl sulfate following acid extraction of histones. Comparisons of histones and non-histones extracted by this method with those obtained by other procedures demonstrate two important advantages of the method; (1) histones obtained by this method are not contaminated by acid-soluble non-histones, and (2) non-histones are not subjected to acid or phenol during extraction. Changes in the distributions of chromatin-associated proteins in different tissues suggest that some species represent regulators of gene action.


Exploration of the neck for hyperparathyroidism. A step-by-step plan for exploration of the neck for hyperparathyroidism has evolved during a 15-year experience with 148 patients at the Toronto General Hospital. Selective catheterization of the veins of the neck with parathormone immunoassay has been a valuable aid in preoperative localization and is particularly helpful in those patients who have had a previous neck dissection. The success of the exploration is based on a practical knowledge of the embryology and anatomy of the parathyroid glands, meticulous dissection and the identification of all glands. The "angle of identification", as described, has been helpful in operative localization. The parathyroid gland or glands that are grossly abnormal are excised, and the remaining glands biopsied and examined by an experienced pathologist, expert in quick-section techniques.


Hyperparathyroidism-an unusual case.

Hyperparathyroidism-an unusual case. A case of primary hyperparathyroidism is described that presented unusual features. The patient had been treated previously for squamous cell carcinoma of the lung and therefore it was necessary to exclude ectopic hyperparathyroidism. The results of the tests generally used to distinguish primary from ectopic hyperparathyroidism-the serum chloride level, the serum calcium response to cortisone suppression and the absence of residual or recurrent tumour-suggested that the patient did have the primary form of the disorder. Exploration of the neck revealed no parathyroid tumour or hyperplasia, and the mediastinal parathyroid adenoma was localized only by determination of parathyroid hormone in the venous drainage from the neck and mediastinum.


Comparison of provincial surgical rates in 1968. Studies have been made of age- and sex-adjusted 1968 provincial rates for individual surgical procedures. For elective and discretionary operations, such as tonsillectomy and adenoidectomy, hysterectomy, vein stripping and lens extraction, provincial rates varied by approximately 100 percent. Newfoundland, with few surgeons and hospital beds, had the lowest discretionary rates, and Alberta, which ranked hgih both for surgeons and hospital beds, the highest. Interprovincial differences were of smaller magnitude for non-discretionary surgery (radical mastectomy, cesarean section, colectomy., lobectomy, etc.). Ther were statistically significant correlations between numbers of surgeons and elective and discretionary surgical rates and between numbers of hospital beds and bed use in the provinces. The effect of disease prevalence on discretionary surgical rates was minimized because the rates were age- and sex-adjusted. Provincial organization of, and payment for, medical services has been similar. Hence, the ratio of surgical personnel to population in each province is postulated as a major determinant of the differing interprovincial rates.


Experimental left gastric vein diversion. Six years' laboratory work resulted in devising a satisfactory experimental technique of direct left gastric-left renal diversion for esophagogastric venous decompression. The procedure was accomplished by interposing an autogenous vein graft between the distal transected left gastric and left renal veins, employing mechanical stapling for the venous anastomoses. A refined technique of left gastric caval diversion provided uniform late patency in a suitable animal model.


Effect of intracranial pressure on cavernous sinus flow: a cadaver model. Through an artificially created carotid-cavernous fistula, the cavernous sinus of four suitable cadavers was perfused at a constant input pressure of 50 cm H20 at various levels of intracranial pressure. The flow rate fell 25% at an intracranial pressure of 50 mm Hg but could not be totally arrested even at the extremes of intracranial hypertension. The authors conclude that the cavernous sinus in man is not collapsible.


Repair of radiation damage in Lewis lung carcinoma cells following in situ treatment with fast neutrons and gamma-rays. Lewis lung tumor cells were irradiated with 60Co gamma-rays or cyclotron-produced neutrons in situ as solid s.c. tumors or in vitro as single cell suspensions. Cell survival was assayed by colony formation both in vitro in soft agar and in the lungs of isogeneic recipient mice. Survival curve characteristics measured in vitro were: Do = 111 rads, Dq = 342 rads, n = 22 for gamma-rays, and Do = 61 rads, Dq = 46 rads, n = 2 for neutrons. In situ, the hypoxic fraction was 0.36. Irradiation in situ gave, for the hypoxic subpopulation, Do = 315 rads for gamma-rays and Do = 91 rads for neutrons. The oxygen-enhancement ratio for gamma-rays was 2.8 and for neutrons was 1.5. Using the split-dose technique, in which two equal doses were administered, separated by 4 hr chronically hypoxic tumor cells repaired sublethal damage, assayed by leaving tumor cells in situ up to 24 hr posttreatment, could not be detected after neutrons, but after gamma-rays it was observed as a 3- to 6-fold increase in survival. The repair of potentially lethal damage increased the relative biological effectiveness of neutrons from 3.7 at a survival level of 5% when assayed immediately after treatment to 4.7 when assayed 6 to 24 hr after treatment. These observations, primarily limited to the chronically hypoxic subpopulation of tumor cells, suggest that decreased repair of potentially lethal damage as well as sublethal damage may be an important radiobiological difference between the effects of high and low linear energy transfer radiation.


Funding: grants or contracts? A survey of cancer scientists. The members of the American Association for Cancer Research (AACR) were polled for their reactions to the current controversy of funding by research grant versus that by the research contract and to the nature of the scientific review appropriate for the evaluation of both types of applications. About 50% of the members responded; 97% of these felt that additional basic knowledge was absolutely essential or probably essential for the successful pursuit of the goals of the National Cancer Program. Eighty % of the respondents concluded that the funding of such research programs should be derived largely from grants rather than contracts. Most of the participants agreed that there should be a major rather than a minor redistribution of funds toward grants and away from contracts. About 50% of the respondent AACR members currently are being supported by research grants, about 10% receive contracts only, and another 25% of the members have both types of funds at their disposal. The group of contract-supported scientists felt less critical of contracts, but about one-half of that group also wished to see more funds going into the grant rather than into the contract programs for the additional support of basic research. There was considerable agreement among all groups of respondents that the review process for contracts should make much more active use of extramural peer evaluation to eliminate scientific inequities now existing between the two instruments for funding of work in cancer research.


Pharmacological studies with vinblastine in the dog. Tritiated vinblastine was prepared by catalytic exchange and its metabolism was studied in dogs. Plasma levels of drug fell in biphasic mode with initial and secondary phase half-lives of 17 to 38 min and 3 to 5 hr, respectively. Between 28.6 and 79.1% of plasma tritium was precipitable with cold trichloroacetic acid and thus was presumably protein bound. Blood leukocytes had levels of intracellular tritium between 2.4 and 11.8 times those of the coincident plasma samples. Over a 9-day period, urinary excretion accounted for 12.1 to 16.8% and fecal excretion accounted for 30.1 to 36.1% of the administered radioactivity. Ratios of biliary to plasma radioactivity varied between 7.3 and 56.9, with unchanged vinblastine being the mamor component (46.8 to 80.7%) in the bile.


The response of hypoxic B16 melanoma cells to in vivo treatment with chemotherapeutic agents. Survival curves are presented for the treatment of B16 melanomas with a range of single doses of cyclophosphamide (CY), 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU), or 1-(2-chloroethyl)-3-trans-4-methylcyclohexyl)-1-nitro-sourea (MeCCNU). When these four drugs are assessed in terms of the tumor cell kill at the lethal dose to 10% of the mice, MeCCNU is found to be much the most effective, followed by CCNU, and then CY and BCNU together. The superiority of MeCCNU is possibly related to the fact that it seems to be longer lived in the mice than are the other drugs. Combined drug and irradiation experiments have indicated that CY kills both oxygenated and hypoxic cells in the tumor, leaving proportions equal to those in the tumor prior to treatment, whereas BCNU preferentially spares the hypoxic cells. Since hypoxic cells constitute a population of cells that is at a distance from blood vessels, this result suggests that CY treatment of B16 melanomas is not limited by an inability of the drug to diffuse to cells away from blood vessels.


Uptake of 67Ga in the regenerating rat liver and its relationship to lysosomal enzyme activity. The uptake of 67Ga citrate has been studied in the regenerating rat liver over a period up to 72 hr after partial hepatectomy. The concentration of 67Ga was found to be maximal (four times that of controls) 42 hr after hepatectomy. This was shown to be related to lysosomal enzyme activity rather than to specific phases of the cell cycle, there being a highly significant correlation (p smaller than 0.001) with aryl sulfatase activity. In both regenerating and normal rat livers, it was shown that 67Ga uptake is reduced when protein synthesis is inhibited by cycloheximide but is unaffected by inhibition of DNA synthesis by cytosine arabinoside.


Tissue water content and nuclear magnetic resonance in normal and tumor tissues. Pulsed proton nuclear magnetic resonance was used to differentiate between normal and malignant tissues. When the tissue water content varied from 80 to 93%, the tumors exhibited spin-lattice relaxation times (T1) from 0.9 to 1.8 sec. We report also the results obtained on 9-day-old embryos and on liver, brain, and heart from 2-day-old rats. A good correlation between the spin-lattice (T1) and spin-spin (T2) relaxation times and the tissue water content was found for all tissues studied. The relaxation times T1 and T2 and water content in Walker 256 carcinoma and its lymph node metastasis were quite similar.


Chemotherapy of the transplantable acute leukemia L5222 in rats. This study presents results of single-drug and combination chemotherapy of the transplantable acute leukemia L5222 in BD IX rats. In leukemia L5222 there is a direct relationship between the number of transplanted cells and mean life expectancy. After single-drug therapy with L-asparaginase, 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), cyclophosphamide, cytosine arabinoside, daunomycin, 6-mercaptopurine, methylglyoxal bis(guanylhydrazone) dihydrochloride, prednisolone, or vincristine, the best therapeutic effect was observed with BCNU and cyclophosphamide. A massive-dose therapy with BCNU repeated twice or a conbination of vincristine with cyclophosphamide or BCNU with cyclophosphamide yielded a high percentage of cures. Morever, leukemia L5222 seems to be suitable for studying the influence of drugs on the proliferation kinetics of leukemia cells.


Combination therapy of mouse leukemia L1210 by 1-beta-D-arabinofuranosylcytosine and 6-[(4-nitrobenzyl)thio]-9-beta-D-ribofuranosylpurine. Nitrobenzylthioinosine (NBMPR), an inhibitor of nucleoside transport, was tested in combination with 1-beta-D-arabinofuranosylcytosine (ara-C) for therapeutic activity against mouse leukemia L1210. NBMPR alone had no activity, whereas therapy with NBMPR and ara-C in combination was significantly better than with ara-C alone. The therapeutic potentiation resulting from the combination of NBMPR and ara-C appeared to be host mediated since NBMPR alone was not toxic to cultured L1210 cells. NBMPR treatment of normal mice increased the plasma half-time of ara-C and decreased rates of urinary excretion of ara-C and 2'-deoxycytidine; however, these effects were not large enough to explain the therapeutic potentiation. Because the drug combination appeared to be no more effective than ara-C alone in therapy of mouse leukemia L1210/TG (a thiopurine-resistant L1210 subline lacking hyposanthine-guanine phosphoribosyltransferase), the host-mediated therapeutic potentiation was attributed in in vivo breakdown of NBMPR to 6-mercaptopurine.


Detection of antibody and complement complexed in vivo on membranes of human cancer cells by mixed hemadsorption techniques. The mixed hemadsorption (MHA) techniques demonstrated antibody and complement fixed in vivo to the surface of human cancer cells. Tumors from 12 cancer patients and normal tissues from 5 cancer patients and 8 patients with cerebrovascular or cardiac diseases were collected from biopsy and autopsy for in vitro testing. Antiserum to human whole immunoglobulins and antiserum to human C3 were used in the MHA techniques. Positive MHA patterns were demonstrated on the surface of cancer cells by both methods. Positive reactions ranged from 12 to 32% in mixed hemadsorption for anitbody detection and from 10 to 34% in mixed hemadsorption for complement component 3 detection. Normal tissues obtained from cancer patients or from patients who died of causes other than cancer rarely exhibited distinct MHA reactivity. Collectively, the data suggest that most human cancers are antigenic in the autologous host and that tumor-associated antigens of cancer cells react in vivo with their humoral antibody to fix complement.


Early and late changes in nonhistone chromatin proteins accompanying rat liver regeneration. Chromatin was isolated from 0.025 M citric acid nuclei of regenerating rat liver at 1,5,18,24, and 48 hr posthepatectomy. The total protein to DNA ratios did not change significantly during this time period. However, 2-dimensional polyacrylamide gel electrophoresis of nonhistone proteins of "Chromatin Fraction II" revealed changes in the amounts of some protein spots. As early as 1 hr after hepatectomy, decreases in size and intensity were detected for protein spots Bp, B24, C18, and CQ, and increases were detected for protein spots CBL and C13. Late changes in size and intensity were found for protein spots BA and CN, which decreased in size and intensity 5 hr after hepatectomy. The spot densities and sizes for most of the nonhistone proteins underwent no significant changes in the course of liver regeneration. The increases and decreases observed in specific protein spots represent an ordered series of changes in a limited number of nonhistone proteins.


Thyroid cell proliferation in rats and induction of tumors by X-rays. There are very few proliferating cells in the thyroid gland of normal adult rats, as measured by the labeling and mitotic index. One-tenth % 4-methyl-2-thiouracil in drinking water induced an exponential increase of thyroid weight after a lag phase of 2 days; the increase continued for 8 days and was followed by a plateau phase. The following sequence of events was found for the number of dividing follicular and stroma cells as well as for DNA synthesis: no significant changes during the 1st 2 days, a sharp increase between the 2nd and 8th days, a decrease between the 8th and 14th days, and an almost constant flow until the 24th day. Three-hundred rads of X-rays given to a nonproliferating thyroid gland induced tumor growth in 25% of the animals 18 months after irradiation. The same dose of irradiation, applied to a proliferating thyroid gland, increased the tumor incidence to 30% when administered in the lag phase, to 75% when administered at the peak of the proliferating phase, and to 62.5% when administered at the plateau phase. Subsequent treatment of irradiated animals with 4-methyl-2-thiouracil enhanced the number and the size of the thyroid tumors and lead to the occurrence of more carcinomas than appeared in animals treated with X-rays only or 4-methyl-2-thiouracil only.


The benzo(alpha)pyrene deoxyribonucleoside products isolated from DNA after metabolism of benzo(alpha)pyrene by rat liver microsomes in the presence of DNA. Rat liver microsomes (induced by 3-methylcholanthrene) were used to catalyze the binding of tritium-labeled benzo(alpha)pyrene to DNA. Enzymic degradation of this DNA to deoxyribonucleosides, followed by separation of the products by Sephadex LH20 column chromatography, revealed two major products. One of these was shown to be the same as that obtained from DNA with benzo(alpha)pyrene bound following treatment of mouse embryo cells in culture with the carcinogen. Neither product resembled those obtained from DNA that had been caused to react with benzo(alpha)pyrene 4,5-oxide (K-region eposide). The aryl hydrocarbon hydroxylase activity of the microsome preparations was determined and related to the extent of microsome-catalyzed hydrocarbon binding. Inhibitors of the enzyme epoxide hydrase increased this binding but caused the loss of one of the two major products. On the basis of the results obtained, a model is proposed of the mechanism of benzo(alpha)pyrene metabolism and DNA binding.


Delayed hypersensitivity to 5-fluorouracil following topical chemotherapy of cutaneous cancers. Skin tests to various common antigens, dinitrochlorobenzene, and 5-fluorouracil (5-FU) were performed on patients being treated for cutaneous neoplasms with topical 5-FU cream. Eleven of 15 patients tested both before and after therapy converted from skin test negative to positive with respect to 5-FU. This conversion correlated with positive dinitrochlorobenzene skin tests and therapeutic cure. The relation between the induction of delayed hypersensitivity reactions to 5-FU following treatment with topical 5-FU and the cure rate for cutaneous neoplasms showed a trend toward correlation.


Evaluation of a cooperative group human myeloma protocol using the MOPC 104E myeloma model. Data are presented on the response rates, maximum rate of cell kill, and survival rates for individual and groups of mice with MOPC 104E myeloma treated with a variety of chemotherapeutic agents and combination regimens used in clinical human myeloma. The tumor immunoglobulin M measurements are used to evalutae the therapeutic effects of drugs. Prednisolone and mescaline, when given as single drugs, showed no therapeutic action and the animals died of tumor as in the controls. The immunoglobulin M values are very similar and ranged between 8,125 and 13,410 mug/mouse. Prednisolone and melphalan given in combination indicated therapeutic effect. 1,3-Bis(2-chloroethyl)-1-nitrosourea-cyclophosphamide-prednisolone combination caused tumor regression but was toxic as shown by the immunoglobulin M values and percentage of survival. The complications and potential uses of this system, which utilized only 40 animals in 64 days, are discussed.


Metabolism of deoxycytidine, thymine, and deoxythymidine in the hamster. The ability of growing and of mature Syrian hamsters to anabolize (to liver DNA) or catabolize (to 14CO2) graded amounts of [2-14C]deoxythymidine (TdR), thymine, or deoxycytidine (CdR) was measured in vivo. Of the three precursors, CdR labeled DNA most efficiently and, as expected, incorporation of all three into DNA was greater in younger animals. The catabolism of [2-14C]CdR to respired 14CO2 was dose dependent and showed no signs whatsoever of saturation, even with the highest dose (greater than 20 mumoles/g liver). In contrast, TdR and thymine were catabolized more slowly and saturation was approached with modest doses. The excretion of CdR in the urine was low and independent of dose, while excretion of TdR and thymine was greater and was dose dependent. Rats tested with an intermediate dose of CdR did not catabolize significant quantities to 14CO2, but did excrete considerably more [C]CdR into the urine than did hamsters. These and other findings suggest that, while the rat and the hamster metabolize thymine (and TdR as well) in a similar fashion, they metabolize CdR quite differently, probably because the hamster has a much higher level of nucleoside aminohydrolase which deaminates CdR and related compounds. Because the human also has a very high level of this enzyme, the hamster appears to be a superior animal model for the study of cytosine-containing compounds intended for human use.


Physiological disposition of pentobarbital in tumor-bearing mice. Pentobarbital depressed macromolecular synthesis in Ehrlich ascites cells in vitro, and this depression was proportional to a decrease in oxygen consumption. However, survival time of animals bearing Ehrlich ascites cells was unaffected by pentobarbital. The acute toxicity of the drug was greatly enhanced by the presence of the tumor. Sleeping time was prolonged in mice carrying the following tumors: Ehrlich ascites, Sarcoma 180 ascites, and Yancy plasma cell solid. Seven-day Ehrlich ascites tumor-bearing animals treated with pentobarbital slept about three times longer than normal mice, but both groups awoke at the same plasma levels of the unbound drug. The plasma half-life of unchanged pentobarbital was about four times as long in tumor-bearing mice as it was in controls. No qualitative difference in catabolism other than rate was detected. Renal excretion of unchanged pentobarbital in tumor-bearing animals was 50% of control animals during the first 4 hr. In tumor-bearing mice the sleeping time of the nonmetabo ble barbiturate, barbital, was identical with that in normal animals. These data suggest that the tumor affected mainly pentobarbital metabolism. Tumor-bearing mice still responded to the pharmacological challenge of phenobarbital with the apparent induction of drug metabolizing enzymes. The prolonged pentobarbital sleeping time in tumor-bearing mice required the development of some type of tumor-host relationship.


Importance of the fecal stream on the induction of colon tumors by azoxymethane in rats. The effect of the fecal stream on intestinal carcinogenesis with azoxymethane was studied in male rats. Colostomies were performed approximately 2 cm distal to the cecum in 50 Sprague-Dawley rats to produce a 20-cm segment of nonfunctional large bowel; an additional 50 animals were left intact. Each of these groups was divided equally and was fed a normal diet or a diet containing 2% cholestyramine by weight. All animals were given azoxymethan s.c. At the end of 7 months all rats were sacrificed. The animals with colostomies developed significantly fewer tumors in the defunctionalized bowel than did intact animals in the same bowel segment. Cholestyramine appeared to increase the tumor yield in the large bowel of the intact animals but had no effect on the number of tumors in the defunctionalized bowel. Further, the intact animals on both dietary regimens developed a greater number of large tumors in the distal 20 cm of bowel. The results show that the fecal stream alters the carcinogenic activity of azoxymethane in the large bowel of the rat. It also appears that the carcinogen can reach its target tissue by a route other than the fecal stream.


Immunocytochemical analysis of cells in the pars tuberalis of the rat hypophysis with antisera to hormones of the pars distalis. The objective was to acquire evidence regarding the secretory capacity of cells in the pars tuberalis of the rat pituitary by the application of immunocytochemical staining. For this purpose the conjugated antibody and immunoglobulin-enzyme bridge techniques were utilized with antisera to the following hormones of the pars distalis: human somatotropin, human thyrotropin, human beta-melanotropin, ovine luteinizing hormone (LH), porcine beta17-39-corticotropin, and beta1-24-corticotropin. Only LH-containing cells were demonstrated. They were exceedingly rare in the cephalic pars tuberalis beneath the median eminence. The frequency of LH-cells was greater in the pars tuberalis associated with the infundibulum and increased distally. LH-cells were most common ventrolateral to the infundibular stem and occurred singly and in clusters. These results indicate that following hypophysectomy the portion of the pars tuberalis that remains in situ has the capacity to secrete only LH of all the pars distalis hormones.


Aspects of furrow membrane formation in the cleaving Drosophila embryo. During blastoderm formation, the Drosophila embryo produces a large area of new membrane to accommodate the simultaneous demands of approximately four thousand newly cleaved cells. The embryo was examined with the electron microscope at various stages during cleavage in order to investigate the high membrane forming capacity of these cells. Embryos were subjected to the histochemical procedure for the demonstration of thiamine pyrophosphatase. The enzyme was present in the cisternae of the endoplasmic reticulum, Golgi bodies and the nuclear envelope. No activity could be demonstrated on the furrow surface or at the furrow tip despite closely adjacent reactive cisternae. It is concluded that the endoplasmic reticulum and Golgi bodies are not major contributors to the new surface. Lamellar bodies were frequently observed in the cytoplasm of all stages examined. The bodies showed a lamellar periodicity of approximately 3.5 nm, and were ultrastructurally similar in appearance after a variety of fixation procedures. The distribution of these bodies was markedly related to the stage of blastoderm formation. Before the commencement of cleavage, lamellar bodies were very prominent within a region 4--5 mum below the cell surface. As cleavage progressed, the bodies became sparse or absent from this region but were apparent at the base of the blastoderm cells or in the sub-blastoderm region, where they were not previously present. Lamellar bodies with leaflets closely associated with, or in apparent continuity with, the cleavage furrow membrane were frequently observed. In these regions the lamellar periodicity was the same as the thickness of the membrane laminae. It is suggested that these bodies play a role in the synthesis of new membrane in the furrow. Intercellular contact specializations between the developing membranes of the furrow were restricted to incipient desmosones and point contacts where the intercellular gap was reduced to 3 nm or less.


Movement of neurosecretory product through the anatomical compartments of the neural lobe of the pituitary gland. An electron microscopic autoradiographic study. Electron-microscope autoradiographs have been prepared from the neural lobes of the pituitary glands of rats which had received intracisternal injections of [35S] cysteine at various times before gland removal. The rate of appearance and disappearance of autoradiographically demonstrable radioactivity in the neural lobe closely paralleled that previously determined, biochemically, for radioactive hormones and neurophysins. Radioactivity was appreciably associated with the undilated parts of neurosecretory axons only during the first few hours after injection of the label. The axonal dilations were subdivided into those in which small vesicles could be seen ("endings") and those in which no small vesicles could be seen ("swellings"). Radioactivity appeared first in "endings" and then in progressively larger and larger profiles of "swellings". It appeared that newly arrived granules were found close to the limiting membrane of the nerve swelling and that as time progressed they moved deeper and deeper into the swelling. On the basis of the results, suggestions were made for an anatomical explanation of the readily-releasable pool of hormone which has been demonstrated pharmacologically.


Chloride cells of larval Notonecta glauca and Naucoris cimicoides (Hemiptera, Hydrocorisae). Fine structure and cell counts at different salinities. The chloride cells of the larval waterbugs, Notonecta glauca L. and Naucoris cimicoides L., ultrastructurally resemble the caviform chloride cells of other aquatic insects as well as those of teleost fish. The predominant features are abundant mitochondria, basolateral plasma membrane infoldings and an apical cavity possessing numerous microvilli. After histochemical precipitation of chloride, dense deposits of silver chloride are present in the apical region. The cuticular area overlying the chloride cells is extremely reduced in thickness and differentiated into a plug-like structure. The 1st instars of Notonecta glauca raised from the eggs at various hypo-osmotic salt concentrations have identical numbers of thoracal chloride cells regardless of the external salinity. The number of cells progressively increases with each moult. However, in corresponding larval stages, there is a significant decrease in the number of thoracal chloride cells relative to increases in the external salinity. These results are consistent with the presumptive involvement of the chloride cells in osmotic hyperregulation.


Chicken gonadotrophins: their effects on the testes of immature and hypophysectomized japanese quail. Five-week old Japanese quail held under short daylengths (8L/16D) were injected daily for two weeks with 20 mug/day of purified chicken luteinizing hormone (LH) or with one of two chicken follicle-stimulating hormone (FSH) fractions. Both intact and hypophysectomized birds were used. After killing, the testes were weighed and fixed for light- and electron-microscopy. Following hypophysectomy testicular weights fell significantly below those of saline-injected controls, indicating that short day conditions are not equivalent to "physiological hypophysectomy", the pituitary-gonadal axis continuing to function at a low level. Chicken LH stimulated differentiation of the interstitium, producing mature Leydig cells containing the full complement of organelles typical of steroid-secretion. In addition it induced some testicular growth caused by spermatogonial division and partial differentiation of the Sertoli cells. This effect was assumed to be a secondary response to androgen secretion but a direct action of chicken LH on the tubules is not dismissed. LH given to intact quail caused greater testicular growth, presumably due to synergism with endogenous hormones. Both chicken FSH fractions induced testicular growth and development; in intact birds the effects were similar to those observed in intact LH-treated birds. In hypohysectomized quail given FSH the testicular weights were greater than in hypophysectomized LH-treated birds and the seminiferous tubules were larger. Sertoli cells were stimulated and pachytene spermatocytes were numerous. Leydig cell differentiation was minimal in all FSH-treated birds, being less pronounced in hypophysectomized than in intact birds.


The effect of oestradiol on the DNA synthesis in neonatal mouse uterus and cervix. (3H)-Thymidine autoradiography was used to study the DNA synthesis in the stroma and epithelium in the uterus proper and the uterine cervix of neonatal mice treated with oestradiol. It was found that in the epithelium of the uterus proper the DNA synthesis is stimulated between 6 and 12 h after injection of oestradiol and decreases again at 18 h. In the stroma of the uterus proper the DNA synthesis is increased 18 h after oestradiol injection. In the epithelium and stroma of the uterine cervix the DNA synthesis decreases from 5 h and is strongly depressed 18 h after oestradiol treatment.


[Semi-automated determination of urinary 17-oxosteroids in a continuous flow system (author's transl)]. A semi-automated determination of 17-oxosteroids in urinary extracts is described using a modified Zimmerman reaction in aqueous phase according to Epstein. In order to eliminate the interfering chromogens a double manifold and a double-beam colorimeter are used in continuous flow. The spectra and the chromogenecity of seven different 17-oxosteroids are presented. The specificity and the reproducibility of this technique are good and the comparison with gas-liquid chromatography and manual Zimmerman reaction shows a good correlation.


An improved benzophenone procedure for the micro-determination of 5,5-diphenylhydantoin in blood. A micro-modification of the benzophenone procedure proposed by Wallace for the determination of 5,5-diphenylhydantoin in blood is described which does not require specialised glassware and avoids the interferance caused by chloroform. In the new procedure 5,5-diphenylhydantoin is extracted from blood with 1,2-dichloroethane, returned to alkali, washed with n-heptane, and oxidised with permanganate in small teflon lined screw capped test tubes containing a layer of n-heptane. The absorbance of the oxidation product (benzophenone) is measured spectrophotometrically on an expanded scale using semi micro cuvettes. The method requires 100-200 mul sample and has a detection threshold of less than 0.1 mg/100 ml.


Putrescine, spermidine, N-acetylspermidine and spermine in the urine of patients with leukaemias and tumors. 1. N-Monoacetylspermidine A and B were isolated from human urine. Structures of the isolated compounds were assigned to monoacetylspermidines by analyses of the products of acid hydrolysis and elementary analyses. The identification was confirmed by comparison of the chromatographic and electrophoretic properties and infrared spectra of the isolated compounds with those of the authentic compounds. 2. The concentrations of urinary polyamines were determined by the combination of ion-exchange chromatography and high-voltage paper electrophoresis. The amount of putrescine fell in the range of 1--6;spermidine, 1--3;spermine, 3--15 and N-acetylspermidines, 2--10 nmoles/mg creatinine for both sexes and different ages. 3. The amounts of urinary polyamines were measured in patients with cancers. The remarkable increase of urinary spermine was observed in blood cancer, while in the other solid cancers the increase of urinary polyamines was not observed.


Modification of an assay for trypsin and its application for the estimation of enteropeptidase. A sensitive method for the estimation of trypsin and enteropeptidase is described. The use of alpha-N-benzoyl-DL-arginine-p-nitroanilide as substrate in combination with the Bratton-Marshall reaction increased the sensitivity of an established method to a degree which permits the determination of enteropeptidase at considerably lower activity levels than heretofore such as those expected in brush border membrane preparations from peroral biopsies of human small intestine.


A study of the renal handling and intestinal absorption of dibasic amino acids in a patient with genotype +/11 heterozygous cystinuria and idiopathic hypercalcuria. A case is reported of a patient with idiopathic hypercalcuria who was referred for investigation of renal calculi. Studies of urinary amino acid excretion in the patient and all living members of his family, as well as studies of endogenous renal clearances of dibasic amino acids and cystine in the patient and his daughter, indicated that the patient had genotype +/11 heterozygous cystinuria. Intestinal perfusion studies showed however that the patient was able to absorb L-lysine, L-arginine normally from the jejunum and ileum. This suggests that a difference exists between renal and intestinal handling of lysine and arginine in cystinuria.


Copper analysis of nail clippings. An attempt to differentiate between normal children and patients suffering from cystic fibrosis. From 39 normal children and 36 patients, suffering from cystic fibrosis (C/F), the copper content of finger nail clippings and toe nail clippings were determined. From this study it can be concluded that, although the patients with cystic fibrosis (C/F) have a higher copper content, the determination of copper in nail samples cannot be used in order to differentiate between normal children and patients. Also higher mean copper concentrations were found in finger nail clippings in comparison with toe nails. This difference was significant for patients with cystic fibrosis.


The reading Miscue Inventory. A promising approach to diagnosis of the reading-disabled. Reading Miscue Inventory--RMI--opens avenues of dealing with the reading process in a scientific and process-oriented fashion which can lead to further refining of the theoretical base upon which reading is founded as well as providing diagnostic information for use by reading specialists. From such information, reading specialists are able to develop remediation on an individualized basis. Appropriate materials, procedures, and practices are developed from the qualitative analysis of miscues rather than from the information provided by standardized achievement or diagnostic reading tests. In sights are gained regarding a child's ability to use the complex process of reading. Remediation of disability, at the appropriate place in the process, is based on assumptions regarding the interrelationship of thought and language. Reading specialists are looking to psycholinguists for theoretical bases upon which to build models of the reading process. Qualitative miscue analysis is based upon recently proposed notions of the psycholinguistic nature of reading. Common concepts and understanding regarding reading and assessment of reading disability may facilitate learning disability teams in their diagnostic and remedial procedures. Until recently, these discussions have occurred mainly in reading-related research and in education circles, but learing disabilities teams are beginning to participate in the refining and clarifying of these notions of assessment and remediation. Regardless of the eventual outcome, the necessity for considering these views is basic to a fuller understanding of ways in which learning-disabled children may be helped.


Complete heart block mimicking seizure disorder. Two patients are described who presented with seizures. Electrocardiogram revealed complete heart block with ventricular asystole lasting four to 10 seconds in each case. These Stokes-Adams episodes were controlled with temporary transvenous pacing. Although there was full clinical recovery in each case, both patients continue to have right bundle branch block (RBBB) and left anterior hemiblock (LAH) 1-1/2 years after their initial presentation.


Observations on methadone withdrawal in 16 neonates. The recent extensive use of methadone for treatment of heroin addiction has taught that, in adults, withdrawal of methadone can produce a withdrawal syndrome similar to heroin. After considerable experience with heroin withdrawal in neonates and in view of controversy and limited studies regarding methadone withdrawal, the study herein reported was undertaken to compare the experiences with both heroin and methadone withdrawal symptoms in newborns. A number of methadone-treated pregnant women deliver their infants at Freedmen's Hospital, affording the opportunity to observe neonates born to methadone-addicted mothers.


A view of Danish pediatric care.

A view of Danish pediatric care. The Danish social system provides competent, accessible, continuous, and coordinated medical care to the pediatric population. The pediatrician does not deal with the routine aspects of well-infant and child care but is rather hospital based and acts on consultant to the generalists in his district. The generalist has the primary role in providing health care to the population in a program where everyone is obliged to participate and no one is excluded or ineligible for any reason. The American obsession with frequent, periodic examinations for infants, children, and young adults does not exist under the Danish pediatric health system. Hospital emergency rooms are used for truly emergent circumstances and not for episodic care.


Medical services at outdoor music festivals. Risks and recommendations. Those planning provision of medical services should consider the type of music, the anticipated size of the crowd, the locale, the available local resources, the length of the festival, the numbers and types of patients to be treated, and especially the potential criminal and civil liability.


Toy safety in hospitals--or beware of parents bearing gifts. Toys are an indispensable component of play therapy for hospitalized children. They can also be dangerous and may result in catastrophic accidents. Extreme care should be exercised in the purchase of "safe toys" for all infants and children in and out of the hospital. The guidelines of the Bureau of Product Safety of the Federal Drug Administration (FDA) dealing with toy safely can be very helpful for child card professionals as well as parents who purchase toys. An explanation of the additional environmental hazards of toys in hospital settings should be an important part of the total in-service training of all personnel who care for infants and children in hospitals. In addition, it might be well for the Division of Product Safety of the FDA to consider criteria for toy safety in hospitals as a separate category in future Bulletins dealing with this problem.


Disposition of methadone in methadone maintenance. Six detoxified opiate addicts housed in a closed metabolic ward received methadone in stepwise increasing doses of 10, 20, 40, and 80 mg/day during 1 month. Four were given 14C-methadone at the lowest dose and again at the highest dose. Of the subjects receiving radiomethadone, 2 excreted the major part of the radioactivity in urine and 2 about equally in urine and feces. In addition to methadone, 7 metabolites were isolated and identified in urine and 3 metabolites in feces. About 75% of the urinary and fecal radioactive metabolites were unconjugated. Urinary excretion of methadone and its major N-monomethylated metabolite accounted for 17% to 57% of the given dose. The ratio of metabolite to parent drug increased in 5 of 6 subjects, and the urinary recovery of unchanged methadone decreased during the period. The results indicate that enhanced demethylation of methadone may occur during oral administration to man.


Etrophine in man. II. Detectability in urine by common screening methods. A single highly euphorogenic dose of etorphine, 100 mug, was administered subcutaneously to 7 nontolerant subjects, and all urine samples were collected for 1 day prior to and 3 days following drug administration. Samples were analyzed for the presence of opiates by radioimmunoassay (Abuscreen) and homogeneous enzyme immunoassay (EMIT), with cutoffs for "ositives" of 40 and 500 ng/ml, respectively. Samples were analyzed for etorphine by thin-layer chromatography (TLC) with iodoplatinate preceded by XAD-2 resin extraction (sensitivity = 0.2 mug etorphine/ml of urine) and by gas-liquid chromatography (GLC) preceded by organic solvent extraction and trimethylsilyl derivatization (sensitivity = 0.1 mug etorphine/ml of urine). The last pre-drug and first two post-drug samples were also analyzed after acid hydrolysis by TLC and after glucuronidase hydrolysis by TLC and GLC. No sample gave a "positive" opiate result in either immunoassay, and no etorphine was detected in the TLC and GLC analyses of any urine sample. Thus, it is unlikely that the abuse of etorphine could be diagnosed by urinalysis using the common screening methods of radioimmunoassay, EMIT, TLC preceded by XAD-2 resin extraction, or GLC preceded by organic solvent extraction and trimethylsilyl derivatization.


Cardiac rate and rhythm changes with atropine and methscopolamine. The effects of methscopolamine bromide (MSB) and atropine were compared in patients prior to elective surgery. After administration of the second dose of MSB (total dose 1.5 mug/kg) all patients exhibited at least 20% increase in heart rate. In contrast, three doses of atropine (total dose 5.3 mug/kg) were required for most patients to attain a 20% increase in heart rate while 2 patients did not attain this heart rate with a dose of 10.6 mug/kg of atropine. Comparison if single injections of MSB and atropine in normal subjects also demonstrated a more reliable dose-response relationship with MSB. Electrocardiographic changes recorded were typical of those reported with anticholinergic agents. Sinus and atrial arrhythmias were more common with atropine and nodal arrhythmias and conduction disturbances were more common with MSB. This study demonstrated that MSB can be reliably substituted for atropine when an anticholinergic drug is needed to increase heart rate. Since MSB has a limited ability to cross the blood-brain barrier, it may be the more desirable drug in patients in whom the central nervous system effects of atropine may be deleterious.


Pharmacokinetics and metabolism of digoxin- and  beta-methyl-digoxin-12aplha-3 H in patients with acute hepatitis. Pharmocokinetics and metabolism of digoxin and beta-methyldigoxin have been studied in patients with acute hepatits after intravenous administration of both H-labeled glycosides. In contrast to digoxin, the rate of decline of radioactivity after administration of beta-methyldigoxin was significantly retarded in patients with acute hepatitis. The increase in plasma concentration after beta-methyldigoxin to patients with acute hepatitis is probably related to decreased demethylation.


Relationship between digoxin concentrations in serum and saliva. The concentration of digoxin in serum and saliva was determined in 18 patients receiving digoxin. Unlike serum, it was necessary to extract saliva with chloroform in order to quantitate digoxin levels accurately. An excellent linear correlation (r = +0.988, p less than 0.001) was observed between the saliva and serum digoxin concentrations. This indicates that saliva digoxin concentrations can be used to monitor digoxin therapy, particularly in patients in whom blood sampling is inconvenient or difficult. The saliva/serum ratio for digoxin concentration was 0.78 plus or minus 0.07 (SD). Since the digoxin binding to plasma proteins is 23%, it is the free drug that is in equilibrium between serum and saliva.


Studies on digitalis. III. Biliary excretion and enterohepatic circulation of digitoxin and its cardioactive metabolites. Simultaneous serum, urine, and bile measurements of digitoxin and its cardioactive metabolites were preformed in 5 cholecystectomized patients with T tube drainage. A 86Rb method was used for serum and urine analysis. The recovery of digitoxin and cardioactive metabolites in two extractions with dichloromethane was 93%; 7% was left in bile. Peak bile concentrations had a mean value of 41.6 ng/ml and were seen after 15 to 60 min. Bile concentration was higher than serum and urine concentration after 24 hr. Mean T/2 of serum elimination was 4.3 days and 8.1 days in 5 control subjects (p less than 0.01). Mean urine concentration T/2 was 10.4 days and 7.2 days in the control subjects (not significant). Mean bile concentration T/2 was 3.5 days. Urinary excretion of digitoxin and cardioactive metabolites was the same in the two groups. The biliary fistula group excreted 22.5% in urine and bile of a dose after 8 days, whereas it was 15.8% in the control subjects. The ratio between the cumulative excretion in urine and bile varied between 1.6 and 2.2. These findings demonstrate that direct interruption of the enterohepatic circulation leads to a marked reduction in serum half-time of digitoxin and cardioactive metabolites, but T/2 is still longer than for other glycosides, indicating that factors other than the enterohepatic circulation are of importance in the slow elimination of digitoxin.


Elimination of procainamide in end stage renal failure. To investigate the effect of end stage renal insufficiency and hemodialysis on the serum half-life of procainamide, 500 mg of procainamide was given orally to control subjects and dialysis patients on interdialysis days. Procainamide was assayed by spectrophotometry and spectrophotofluorometry. Mean half-life in normal subjects was 3.2 hr by spectrophotometry and 3.5 hr by spectrophotofluorometry. Mean half-life in patients was 11.3 hr by spectrophotometry and 16.0 hr by spectrophotofluorometry (p less than 0.001 compared to control subjects). Half-life of procainamide during dialysis in patients given 500 mg of procainamide 1 hr before dialysis was 4.3 hr and 9.6 hr on a nondialysis day (p less than 0.001). Both methods of assay gave higher levels of procainamide when the metabolite, N-acetylprocainamide, was present in serum and the extract allowed to stand in 1 N HCl, but spectrophotometry was less affected. Thus, end stage renal insufficiency greatly prolongs the half-life of procainamide, procainamide is readily dialyzable, and N-acetylprocainamide is hydrolyzed in 1 N HCl to procainamide during routine serum determinations.


Morphology of gastrointestinal effects of aspirin. The effect of acetylsalicylic acid (aspirin) on the ultrastructure of gastric and jejunal mucosa was investigated in patients undergoing gastric surgery and in guinea pigs. The drug caused various degrees of damage to the surface mucous cells in both species perceptible as general signs of cytolysis in situ or as desquanmation. In patients pretreated with aspirin, an increase of secondary lysosomes was noted in the gastric parietal cells but not in the animals. In the intestinal epithelial cells of both species there was a marked increase of multivesicular bodies and the occurrence of transitional stages between the two organoids suggest a functional interrelationship. Since no specific alteration of any cellular organoid was detected, the drug-induced injury is assumed to occur on a molecular level in the cytoplasm. It is concluded that the intracellular concentrations of aspirin in gastrointestinal mucosal cells mechanims but that high drug concentrations may lead to irreversible cell damage.


Pharmacokinetics of sulbenicillin, a new broad-spectrum semisynthetic penicillin. In a pharmacolinetic study on a new semisynthetic penicillin, alpha-sulfobenzylpenicillin, sulbenicillin, serum level, serum half-life, apparent distribution volume, renal clearance, urinary excretion, and metabolism were determined after a 4-gm intravenous dose and compared to that of carbenicillin in 5 patients with normal renal function. In the case of sulbenicillin, the mean serum concentration at 1 hr was 157 plus or minus 25 mug/ml, the mean serum half-life was 70 plus or minus 10 min, the renal clearance was 95 plus or minus 25 ml/min, and the total urinary recovery after 24 hr was about 80% of the dose. The only metabolite detected in the urine was the penicilloic acid derivative, in an amount usually less than 5% of the dose. Serum values, serum half-live, renal clearances, and excretion pattern did not differ significantly from that of carbenicillin. In 8 patients with decreased renal function (creatinine clearance less than 50 ml/min) there was an inverse correlation between creatinine clearance and serum half-life.


Pharmacokinetics of bethanidine in hypertensive patients. The pharmacolinetics of bethanidine-14C was studied in three hypertensive patients. A 25-MG DOSE OF BETHANIDINE-14 C hemisulfate was administered intravenously. Plasma levels of drug were measured over the first 6 hr. In 3 to 4 days, 89% to 94% of the dose was excreted in the urine. Thin-layer chromatography (TLC) and isotope dilution analysis of the urine samples indicated that only intact bethanidine was excreted. Plasma level and urinary excretion rate profiles had miltiphasic characteristics. Estimated half-lives of the terminal phase ranged from 7 to 11 hr. Average renal clearance over the initial 6 hr approached renal plasma flow. In 2 of the patients, renal clearance between 2 and 4 hr after administration was reduced to one-helf that observed during the initial 2-hr period. After single oral administration of a 25-mg dose of bethanidine-14C hemisulfate, 48% of 61% was excreted in urine and 15% to 48% in feces. Peak urinary excretion rates were reached 6 hr following administration. The urinary excretion kinetics of bethanidine during and after repetive oral dosing was also studied. A 25-mg dose was dividied into 12 to 16 equal doses and administered avery 6 hr. A larger fraction of the cumulative dose was recovered in the urine (72% to 74%) than after the single dose, suggesting higher availability at the lower dose. Steady-state urinary excretion rates were achieved in 4 to 7 doses. The steady-state urinary excretion levels were consistent with pharmacolinetic predictions based on single oral dose data. When 2 of the patients were given imipramine for 2 days prior to an oral 25-mg dose of bethanidine-14C hemisulfate, the terminal half-lives of the urinary excretion rate profiles were shorter than those in the same patients not given imipramine.


[Early gastric carcinoma].

[Early gastric carcinoma]. The early gastric cancer can now be diagnosed more frequently by improved methods of roentgenology and endoscopy. At present 16.7 percent of the gastric resections for cancer in our clinic are for early gastric cancers. Indicative clinical symptoms were demonstrable in 25 of our own 27 cases described here. Infiltration of the gastric wall (mucosa- and submucosatype) and lymph-node metastases are important to the prognosis. Intraoperatively, it is essential to localize the tumor unobjectionably and to perform a sufficiently radical resection (generally subtotal gastrectomy and excision of the lymph-nodes). The 5-year-survival-rate totals 70 to nearly 100 percent. Postoperatively controls are necessary for the diagnosis and eventually the treatment of possible recurrence.


[Pyloric stenosis caused by gallstone diseases]. During the 15 year period between 1958 and 1973 the authors operated on 21 cases of pyloric stenosis caused by biliary calculi, occuring in 12 percent of their patients suffering from pyloric stenosis. They call attention to the fact that cholecystectomy made in good time can prevent pyloric stenosis being a complication of neglected cases.


[Head endoprostheses in femoral neck fractures]. By clinical, radiographic and scintigraphic means we attempted to examine, whether the implantation of a simple head-endoprosthesis after fractures of the femoral neck still has to be considered as an appropriate operating method or whether on principle, primary endoprosthesis should be used. The results prove that in suitable cases and with correct technique the head-endoprosthesis still has justification, especially as the "protrusio acetabuli", known only as complication of pure head-endoprosthesis, is mostly avoidable. Reference to the choice of cases, operating conditions and appropriate methods of examination are given.


[Dissecting aneurysm with damage to median nerve]. A case of iatrogenic dissecting aneurysm in the cubital region causing secondary damage to the median nerve is reported. This aneurysm was caused by an attempted venipuncture, in what can be considered a minor procedure. Such a severe complication is very remarkable, for it has not been observed so far even in much more potentially hazardous procedures as, for instance, in the numerous arterial punctures in the cubital region being daily performed for retrograde angiography of the cerebral vessels. In this case the reduced blood coagulation is discussed as a causing factor of this unusual complication.


Immunity in colonic tumor patients after operation: determination by leukocyte-migration inhibition. Tumor immunity in 31 colonic tumor patients was studied by determining in vitro leukocyte-migration inhibition. In tissue culture 30 per cent inhibition of patient leukocyte migration by tumor antigen could be demonstrated in half of the patients, suggesting that they were immune. This immunity was not related to the clinical or pathologic stage of the disease and varied in different patients. It was transient in some and persistent in others. In addition, a positive response at a fixed time after operation was not found.


Parenteral iron in treating surgical patients and the importance of routine evaluation of serum iron-binding capacity: report of three cases. We have endeavored to show the importance of iron deficiency and to cite a few examples of the systemic effects of such deficiency without anemia. We have shown the need for large-scale screening of patients for iron deficiency and suggest that serum iron and total iron-binding capacity determinations will adequately furnish the vehicle for this purpose. The modes of iron therapy are discussed with the belief that for the surgeon parenteral iron is the best vehicle to replace the deficiency with the least risk.


Effect of thyrocalcitonin on calcium concentration in liver of intact and thyroparathyroidectomized rats. Thyrocalcitonin (TCT; 80 MRC mU/100g) administered SC to intact and thyroparathyroidectomized rats, caused a significant decrease in serum calcium, while increasing the liver calcium concentration. The accumulation of calcium in the liver, induced by TCT, was potentiated by the injection of calcium in both intact and thyroparathyroidectomized rats. In thyroparathyroidectomized rats, the administration of imidazole (30 mg/100 g) produced a significant increase in liver calcium and a corresponding fall in serum calcium. The rise in liver calcium produced by TCT was slightly potentiated by prior administration of imidazole, but was not enhanced by prior administration of theophylline (12 mg/100 g) which caused a significant elevation of serum calcium. These results suggest that the action of TCT on liver calcium does not involve cyclic AMP.


Cycloleucine transport in isolated rat thymocytes: in vitro effects of triiodothyronine and thyroxine. Thymocytes obtained from suckling or young adult rats were used as a model system to study the action of thyroid hormones in vitro. In this tissue, L-triiodothyronine (T3) increased the uptake of the non-metabolizable amino acids, alpha-aminoisobutyrate and cycloleucine. A detectable effect of T3 on the uptake of cycloleucine was seen at a concentration of 0.1 muM and maximum effects were seen at 20 muM. Thyroxine (T4) also increased cycloleucine uptake with about one-third the potency of T3, and this effect could not be ascribed to conversion of T4 to T3. In contrast, L-monoidotyrosine and L-diiodotyrosine were without effects on transport. Kinetic studies indicated that T3 enhanced uptake by inhibiting amino acid efflux; no effect was seen on influx. The effect of T3 on amino acid uptake was evident within 1 min, and was not inhibited by either prior treatment of the cells with cycloheximide or by lowering the incubation temperature from 37 to 24 C. In other studies, when T3 was injected into rats in vivo at a dose of 20 mug/100 g, the uptake of cycloleucine was enhanced in thymocytes obtained 1 h later. These data suggest that thyroid hormones can directly influence amino acid transport in rat thymocytes. This effect is prompt, is independent of new protein synthesis, and may reflect a direct interaction with specific components of the cell membrane.


Arginine vasotocin in the rabbit subcommissural organ. Subcommissural organs of young and mature rabbits were analyzed for their content of arginine vasotocin by radioimmunoassay. Younger animals had significantly greater quantities of this peptide. There was no detectable arginine vasopressin or oxytocin in subcommissural organ extracts. It is concluded that the subcommissural organ represents, in addition to the pineal and the fetal neurohypophysis, another significant source of arginine vasotocin in the mammalian central nervous system.


Thyrocalcitonin injection to rats increases the liver inorganic phosphate. The mechanism of the hypophosphatemic effect of thyrocalcitonin (TCT) is uncertain and may involve removal of inorganic phosphate (P1) from the plasma by heretofore unidentified organs. Rats, 192 g. thyroparathyroidectomized 16-24 h earlier, received 100 MRC mU TCT or vehicle (V) SC and 100 muCi 32PO4 iv. One h later plasma and tissue samples were collected. Compared to rats which received V, TCT-treated rats had lower plasma P1 (-0.36 plus or minus 0.08 mumol/ml, P smaller than 0.001) and Ca, higher liver P1 (+0.66 plus or minus 0.19 mumol/g, P smaller than 0.01) and unchanged red blood cell and rectus abdominus P1. TCT had no significant effect on the amount of perchloric acid soluble phosphate in any of the tissues. TCT-treated rats had lower plasma 32P1 (cpm/ml) and higher liver 32P1 (cpm/g) with no significant changes in specific activity (SA). TCT caused a small decrease in muscle 32P1 SA. There were no significant changes in the 32P (cpm/g) or 32P SA of the acid soluble phosphate fraction of any of the tissues. The results suggest that a portion of the hypophosphatemic effect of TCT can be accounted for by the increased P1 level in the liver.


Catechol estrogen formation in the central nervous system of the rat. Conversion of estradiol and estrone to catechol estrogens by rat hypothalamic tissue but not by the cerebral cortex was demonstrated from the incubation of these tissues with estradiol-2-3H and estrone-2-3H and monitoring the incorporation of tritium into water. Direct evidence for this transformation was obtained by isolating the labelled phenazine derivative of 2-hydroxyestrone after hypothalamic incubation with estrone-4-14C.


The effect of fetal hypophysectomy on placental biosynthesis of progesterone in rhesus. Placental slices from intact rhesus fetuses were incubated without added substrate. The incubated tissue levels of progesterone (P4) differed according to the sex of the fetus. Slices from female placentas contained significantly more P4 than did those of males. The addition of pregnenolone (P5) to the incubation media caused tissue levels of P4 to increase 1.5 to 3 times control tissue levels for both female and male placentas. Moreover, the sex difference in P4 biosynthesis was eliminated by adding P5 to the incubations. Since control incubations of male placental tissue produced less P4 than those of females, the net increase in P4 synthesis with added P5 was greater for male than female placental tissue. These observations indicated that the step(s) in P4 biosynthesis which were affected by the fetal genotype lay between cholestrol and P5. Incubation of placental slices derived from decapitated fetuses secreted significantly less P4 into the incubation medium than those of intact fetuses. Moreover, the sex difference in the media content of P4 was eliminated. However, decapitation did not eliminate the sex difference in the tissue content of P4 during control incubations. When P5 was added to the incubations, media and tissue levels of P4 increased significantly over control levels for placentas from both sexes. However, the addition of P5 to the incubations from decapitated males did not restore P4 production in the tissue or the medium to levels observed for intact males. However, this did occur when P5 was added to incubations from decapitated females. It appears that fetal decapitation decreased cholesterol side chain cleaving activity and delta5-3beta-hydroxy-steroid dehydrogenase content of the placenta. These data indicate that hormones of fetal origin may control P4 production by the placenta.


Regulation of pineal rhythms in chickens: refractory period and nonvisual light perception. Pineal serotonin N-acetyltransferase activity and melatonin content exhibit marked daily rhythms in chickens; peak values occur during the period of low locomotor activity which coincides with dark in a 24-h light-dark regime. We studied the regulation of these daily rhythms by measuring pineal serotonin N-acetyltransferase activity, hydroxyindole-O-methyltransferase (HIOMT) activity, and melatonin content in experiments in which birds were exposed to light-to-dark and dark-to-light transitions at atypical times. We observed that there is a refractory period for dark-initiation of the increase in pineal N-acetyltransferase activity and melatonin content. We also learned that a dark-to-light transition causes a rapid decrease in dark-elevated pineal N-acetyltransferase activity and melatonin content. The rapid decrease occurs in blinded chickens as well as sighted ones. HIOMT activity did not change consistently in any of the experimental treatments.


The interaction of castration and photoperiod in the regulation of hypophyseal and serum gonadotropin levels in male golden hamsters. Luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were measured in intact and castrate adult male hamsters maintained on photostimulatory (LD 14:10) and non-photostimulatory (LD 6:18) light:dark cycles to assess the interaction of photic stimuli and gonadal hormones on pituitary gonadotropin release. Immunoreactive serum LH and FSH levels increased 1.6- and 8-fold respectively, within 3 days after photostimulated hamsters were castrated. In contrast, castration failed to alter serum LH concentration and had only a slight, if any, effect on FSH concentration in hamsters exposed to nonstimulatory photoperiods that induced testicular atrophy. In a second experiment, male hamsters previously maintained on LD 14:10 were castrated, transferred with intact animals to LD 6:18, and killed periodically over 60 days. In intact animals, pituitary content and serum levels of LH and FSH declined substantially during exposure to the non-stimulatory LD 6:18 cycle. In castrated animals, serum LH and FSH levels which had increased 2- and 8-fold in response to the castration eventually declined to about the levels found in the intact initial control animals. In contrast to serum gonadotropins, the increased hypophyseal content of LH and FSH following castration was not reduced during exposure to LD 6:18. Exposure to nonstimulatory photoperiods does not alter the increased hypophyseal LH and FSH content observed after castration. However, our results indicate that exposure to short days renders the hypothalamic-hypophyseal neuroendocrine system governing gonadotropin release relatively insensitive to gonadal steroid hormone feedback.


Differential responses of prelactating and lactating mammary gland to similar tissue concentrations of progesterone. Serum and mammary tissue concentrations of progesterone and 20alpha-hydroxy-4-pregnen-3-one (20alpha-OHP) were measured by competitive protein-binding assays and gas-liquid chromatography, respectively, in pregnant and lactating rats. The concentration of progesterone in mammary tissue of pregnant rats closely paralleled the serum concentration, particularly when tissue concentration was expressed as ng/mg DNA. The variability in tissue progesterone on the last day of pregnancy was relatively great, but there was a good inverse relation between the appearance of lactose and the progesterone concentration. Serum progesterone levels declined to their lowest values at 1-3 days of lactation (10 plus or minus 1 ng/ml); the tissue concentration declined even more rapidly after parturition. The tissue 20alpha-OHP concentration, which was more closely related to serum progesterone among animals than to serum 20alpha-OHP, remained high after parturition suggesting that the presence of 20alpha-OHP has no effect on lactogenesis and that progesterone is decreased in the tissue by 20alpha-reduction. Following postpartum ovulation, serum progesterone increased to 74 plus or minus 6 ng/ml at 6-9 days of lactation; tissue progesterone also increased to levels found in rats pregnant 14-19 days, yet no change in lactose content of the glands of suckled rats occurred, and the biosynthetic capacity in terms of the RNA/DNA ratio increased. Serum 20alpha-OHP also rose, but the tissue concentration was unchanged, suggesting that saturating levels were present throughout pregnancy and lactation. Since no rapid increase in DNA was associated with lactogenesis, differentiation of nonsecretory parenchymal cells into daughter cells with the secretory capacity must occur earlier in pregnancy. Progesterone, therefore, must inhibit lactogenesis by preventing expression of the genetic potential of daughter cells. Once differentiation has been completed, however, the presence of progesterone in the tissue has no effect on the biosynthesis of milk constituents.


Time course of the effect of adrenalectomy on transcortin binding characteristics: appraisal of different methods of calculation. The time course effect of adrenalectomy on the transcortin-corticosterone association constant (KT), its concentration of binding sites (ST) and the corresponding binding constant (SAKA) of albumin has been investigated by equilibrium dialysis at 37 C on plasma samples collected 0, 3, 6, 12, 24, 48, 96 and 144 h after adrenalectomy in male adult rats. The data have been analyzed by 3 different methods: a graphical Scatchard analysis using bound over unbound versus bound as representation axes and 2 least squares minimization methods in which axes were, respectively, bound over total versus mass of corticosterone added and total versus unbound. Confidence regions can be computed in the last two methods allowing the statistical comparison of the different dialysis experiments. Results obtained by the three methods are quite similar and led to the conclusions that following adrenalectomy KT was constant while ST varied. The observed pattern was a decrease until the first 24 h followed by a rise reaching almost twice the control value at 144 h. It would also appear that SAKA decreases slightly within 24 h following adrenalectomy.


Hormonal regulation of isoenzymes of N-acetyl-beta-glucosaminidase and beta-galactosidase during spermatogenesis in the rat. Isoenzymes of beta-galactosidase and of N-acetyl-beta-glucosaminidase were assayed during development of rat testis and as a function of hormonal treatments. Isoenzyme 1 of beta-galactosidase was highest in specific activity in the 4-day-old testis, at a point when Sertoli cells and gonocytes were the predominant cell type. Beta-galactosidase II, previously shown to be associated with the sperm acrosome, was undetectable through the spermatocyte stage of development, but increased in specific activity during the formation of spermatids. The specific activities of isoenzymes I and II of N-acetyl-beta-glucosaminidase increased markedly in association with the formation of spermatogonia and spermatocytes, and then declined with the appearance of spermatids. Following hypophysectomy of rats at 26 days of age or in adulthood the specific activities of the lysosomal enzymes beta-galactosidase I and N-acetyl-beta-glucosaminidase I and II increased markedly, while the acrosomal beta-galactosidase II was undetectable. The normal patterns of isoenzyme distributed were restored completely by administration of LH and FSH or testosterone to hypophysectomized animals. These results thus demonstrate specific patterns of isoenzyme concentration during spermatogenesis. Formation of the acrosome in developing spermatids is associated with the induction of new forms of beta-galactosidase (isoenzyme II) and N-acetyl-beta-glucosaminidase (sperm isoenzyme). These molecules appear to be specialized forms which may participate in fertilization, and their induction is dependent upon the actions of gonadotropins or testosterone.


Cellular localization of 5alpha-reductase and 3alpha-hydroxysteroid dehydrogenase in the seminiferous tubule of the rat testis. Seminiferous tubules isolated from normal adult rats converted (14C) testosterone to (14C) androstanediol and (14C) androstenedione as the major metabolites; (14C) dihydrotestosterone and (14C) androsterone were produced in lesser amounts. Tubules from immature rats (26-28 days of age) converted a higher proportion of (14C) testosterone to 5alpha-reduced products than did tubules from adult rats. Spermatocyte-enriched preparations contain 5alpha-reductase. The lower level of 5alpha-reductase activity in spermatid-spermatocyte preparations indicates that this enzyme is low or absent in spermatids. Sertoli cell-enriched preparations contain 5 alpha-reductase and a high level of 3alpha-hydroxysteroid dehydrogenase. The results show that spermatocytes and Sertoli cells have the capacity to metabolise (14C) testosterone to 5alpha-reduced products; dihydrotesterone is the major product formed by spermatocytes, whereas in Sertoli cells further metabolism to 5alpha-androstane-3alpha, 17beta-diol occurs.


Glucocorticoid inhibition of sulfate incorporation by cartilage of normal rats. The effects of dexamethasone, cortisol, and deoxycorticosterone on sulfate incorporation by cartilage of normal rats were determined. In vivo dexamethasone was a more potent inhibitor than cortisol, and deoxycorticosterone in the dose tested was ineffective. In vitro dexamethasone and cortisol were inhibitory in concentrations of 10-8 and 10-7M, respectively, when sulfate incorporation was measured after 24 h of incubation. Dexamethasone was at least 10 times as potent as cortisol. Deoxycorticosterone was inhibitory in a concentration of 10-4M. It is concluded that inhibition of mucopolysaccharide synthesis in cartilage of normal rats by adrenal cortical steroids or their analogs, both in vivo and in vitro, is correlated with glucocorticoid activity.


Microtubules and the intracellular conversion of proparathyroid hormone to parathyroid hormone. The effects of several compounds which interfere with cellular microfilaments and microtubules on the conversion of proparathyroid hormone (ProPTH) to parathyroid hormone (PTH) were examined in slices of bovine parathyroid slices incubated in vitro with 3H-leucine. Inhibitors of microtubular function, vinblastine and colchicine, increased the ratio of 3H-labeled ProPTH to PTH in the tissue by 2- to 3-fold. Cytochalasin B, an inhibitor of microfilaments, was without effect. Concentrations of colchicine as low as 10-6M maximally increased the ratio of ProPTH to PTH, whereas lumicolchicine, an analogue of colchicine which does not affect the function of microtubules, had no effect at concentrations as high as 10-4M. The increased ratio of ProPTH to PTH was due partly to a stimulation by vinblastine and colchicine of the incorporation of 3H-leucine into ProPTH but not into general protein. However, after short incubations of parathyroid tissue with 3-H-leucine, the amount of 3H-labeled PTH was less in colchine-treated incubations than in control incubations. In the presence of vinblastine, after a 20-min incubation of parathyroid slices with 3H-leucine and vinblastine, ProPTH was not completely converted to PTH by an additional 90-min "chase" incubation with unlabeled leucine while a 20-min "chase" incubations is sufficient to convert essentially all the ProPTH to PTH in control incubations. These data suggest that the increased ratio of ProPTH to PTH is also due to a partial inhibition of the conversion ProPTH to PTH by vinblastine and colchicine. The data are consistent with the hypothesis that microtubules facilitate the transport of newly synthesized ProPTH to its intracellular site of cleavage to PTH.


An inhibitory influence of granulosa cells and follicular fluid upon porcine oocyte meiosis in vitro. Isolated oocytes will resume meiosis spontaneously in vitro whereas follicle-enclosed oocytes will remain in the dictyate stage when cultured unless they have been exposed to gonadotropins in vivo or in vitro. To examine the source of the follicular inhibitory influence, porcine oocytes have been cultured alone, with hemisections of follicle wall, granulosa cells, or with follicular fluid. Oocytes isolated from medium-sized (3-5 min) follicles resumed meiosis when cultured; 77.5 plus or minus 3.4 percent matured beyond the dictyate stage. When oocytes were cultured in the presence of follicle wall hemisections of medium and large (6-12 mm) follicles, the percentage of maturing oocytes was significantly reduced. The maturation of oocytes cultured in a medium containing 50 percent follicular fluid from small or large follicles was significantly inhibited. Resumption of meiosis was completely inhibited by co-culture of isolated oocytes with 10-7 granulosa cells from small, medium or large follicles. Addition of serially diminishing amounts of granulosa cells from 10-7 to 10-4 cells reduced the inhibitory influence. It is concluded that the granulosa cells are responsible for the maintenance of the oocytes in the dictyate stage within the follicle. The granulosa cells appear to exert their inhibitory influence upon meiosis by secretion of a chemical message into follicular fluid.


Hypothalamic 24-hour rhythms in histamine, histidine, decarboxylase and histamine-N-methyltransferase. Twenty-four hour rhythms of histamine (H), its biosynthetic enzyme, L-histidine decarboxylase (EC 4.1.1.22, HD) and its inactivation enzyme, histamine N-methyltransferase (EC 2.1.1.8, HMT) were assayed in the 63-day-old male rat hypothalamus using a modified single isotopeenzyme microassay. H was found to have a 24-h rhythm with a maximum concentration at 11:00 h (lights on 10:00-22:00 h) and a minimum at 23:00 h. In vitro HD and HMT activities were also found to have 24-h rhythms with peak activites in both occurring at 04:00 h. HMT also had a broad secondary peak throughout the day, whereas HD activity during the day was dominated by a mid-light trough at 16:00 h. These rhythms are considered significant for evaluation of the control and physiological role of histamine in the mammalian hypothalamus.


Steroid hormone-responsive, isolated endometrial cells. Addition of estradiol-17beta in vitro to suspensions of isolated endometrial cells resulted in significant effects on glucose, water and electrolyte metabolism. Cells were prepared from uterine tissues of ovariectomized rats. In part, the procedures involved incubation with collagenase in Ca2+-, Mg2+-free, phosphate-buffered mammalian Ringer's solution, followed by restoration of divalent cations before gentle scraping of the endometrium from the underlying smoothmuscle. Cells were then disaggregated, washed, separated from coarse and fine debris, and incubated in an enriched medium for 2 h before the start of all experiments. Cellular integrity was established by measurement of electrolyte contents and by dye exclusion methods. Substantial production of 14CO2 from glucose-U-14C by the cell suspensions provided further evidence of cell viability. Estradiol-17beta, 10-9M, elicited significant increments in sodium and water contents within 2 h. Addition of estradiol-17beta, but not the alpha-epimer, also resulted in a significant increase in the yield of 14CO2 as early as 1.5 h, peaking at 2 h. The responses were dose-dependent between 10-10M through 10-8M. The stimulatory effect of estradiol-17beta at 10-9M was abolished in the presence of 3 times 10-6M cortisol or by cellular homogenization. Epithelial cells isolated from rat urinary bladder responded significantly to 6 times 10-9M aldosterone but not to estradiol-17beta, demonstrating specificity of the target site. These data lend further support to the suggestion that a primary action of estrogen in its target cell involves specific changes in the ionic and biochemical profile of the cytoplasm which may ultimately be communicated to the nucleus.


Steroid levels in follicles and the plasma of hens during the ovulatory cycle. Laying hens were killed at hourly intervals during the 26-h laying cycle. The 3 largest follicles of the size hierarchy were removed and plasma samples were obtained from the same hens. The follicle walls and the plasma were assayed by RIA for estrogen (E), progesterone (P4), and testosterone (T). The data show that for E and T there is an inverse relationship between follicle size and hormone concentration. There is no such difference for P4. Both E and T show a significant drop immediately after ovulation; P4 does not. Both T and E concentrations rise significantly and synchronously at about 4 and at 8 h after ovulation. The OAAD assay of plasma shows a peak of LH about that time. P4 does not show such a rise in concentration. About 8 to 10 h prior to the next ovulation T concentration in all 3 follicles begins to rise, P4 rises only in the largest follicle and E only in the smallest. All three steroids reach highly significant peaks about 4 to 6 h prior to the next ovulation. Both OAAD and RIA detect plasma LH peaks at about that time. Plasma E and follicle E peak synchronously at 4 to 8 h; T peaks occur at 10 to 12 h after ovulation and are asynchronous with the T follicle peaks. All 3 steroids begin to rise in the plasma about 10 h prior to the next ovulation and all 3 peak together about 4 to 5 h prior to ovulation. The present data do not allow to distinguish between rates of steroid synthesis and their release into the plasma. Whether the steroid peaks occurring in both follicles and plasma shortly before the next ovulation are caused by LH or are the cause of its release remains to be determined.


Effect of LH injection on plasma and follicular steroids in the chicken. During the 26-h laying cycle of the hen the three sex steroids, progesterone (P4), estrogen (E), and testosterone (T), show peaks and valleys in both the follicle wall and the peripheral plasma. Furthermore, for E and T (but not for P4) at comparable times, there is an inverse relationship between the size of the follicle and the concentration of these hormones. Shortly before ovulation all three steroids peak in synchrony, at other times only E and T rise and fall together while P4 shows no change. The question arose whether similar changes in steroid synthesis can be produced by exogenous LH. A single injection of 25 mug of ovine LH was given to laying hens at a time of the laying cycle when no natural peaks of the steroids are seen. The hens were killed at 5, 30, or 90 min after injection and the walls of the three largest follicles as well as the plasma of these hens were assayed by RIA for P4, T, and E. Both the follicle walls and the plasma showed highly significant (P smaller than 0.005) increases in T and P4 but not in E, even though E does show significant changes during the normal cycle. The reason for this difference in response of the steroids is being investigated. The 3 largest follicles respond to exogenous LH in the same size order as do untreated hens during the cycle. The conclusion is drawn that peaks of T and P4 normally seen are due to endogenous releases of LH while E seems to be controlled differently.


Optimal statistical design of radioimmunoassays and competitive protein-binding assays. The statistical analysis of radioimmunoassay and competitive protein binding assay data is complex. Because the response variable (percent counts) is not lineraly related to log dose, a logit transformation of the response variable usually is performed to permit linear regression analysis. This transformation induces marked heterogeneity of variance, so that iterative weighted regression programs have been used to achieve the best standard curve and the most precise dose estimates of unknowns. In this study several parameters of assay design are investigated in order to establish those designs yielding antigen concentration estimates of highest precision as well as estimates of comparable precision by either simple linear regression analysis or by the more complex weighted regression technique. Unknown estimates of highest precision are obtained when 1) the present counts of the standard doses covers a range of approximately 80 percent to 20 percent, 2) the number of standard dose levels is eight or more, 3) the number of replicates at each dose level is two or more, and 4) the percent counts of the unknowns also are within the range 80 percent ot 20 percent. Under these conditions, also, simple linear regression yields unknown estimates of comparable precision to weighted regression and therefore may be safely used.


Polymorphism of centromeric heterochromatin and its relationship to chiasma formation in the paracentromeric regions of murine chromosomes. A hypothesis suggesting an evolutionary significance of centromeric heterochromatin polymorphism, which presumed the suppression of genetic crossing-over in the paracentromeric chromosome regions, was experimentally tested. The data obtained do not support the hypothesis; the frequency of paracentromeric chiasmata in two inbred mouse strains (differing in centromeric heterochromatin pattern) is not higher than in their F-1 hybrids. C-bands staining of centromeric heterochromatin allowed the distinction of two types of paracentromeric chiasmata. One of them--pseudochiasmata--is probably the consequence of centromeric heterochromatin affinity rather than the result of the preceding crossing-over.