A backflow of electrons around photosystem II in Chlorella cells.

A study was made with a modulated oxygen electrode of the effect of variations of oxygen concentration on photosynthetic oxygen evolution from algal cells. When Chlorella vulgaris is examined with a modulated 650 nm light at 22 degrees C, both the oxygen yield and the phase lag between the modulated oxygen signal and the light modulations have virtually constant values between 800 and 120 ergs . cm-1 . s-1 if the bathing medium is in equilibrium with the air. Similar results are obtained at 32 degrees C between 1600 and 120 ergs . cm-2 . s-1. Under anaerobic conditions both the oxygen yield and the phase lag decrease if the light intensity is lowered below about 500 ergs . cm-2 . s-1 at 22 degrees C or about 1000 ergs . cm-2 . s-1 at 32 degrees C. A modulated 706 nm beam also gives rise to these phenomena but only at significantly lower rates of oxygen evolution. The cells of Anacystis nidulans and Porphyridium cruentum appear to react in the same way to anaerobic conditions as C. vulgaris. An examination of possible mechanisms to explain these results was performed using a computer simulation of photosynthetic electron transport. The simulation suggests that a backflow of electrons from a redox pool between the Photosystems to the rate-limiting reaction between Photosystem II and the water-splitting act can cause a decrease in oxygen yield and phase lag. If the pool between the Photosystems is in a very reduced state a significant cyclic flow is expected, whereas if the pool is largely oxidized little or no cyclic flow should occur. It is shown that the effects of 706 nm illumination and removal of oxygen can be interpreted in accordance with these proposals. Since a partial inhibition of oxygen evolution by 3-(3.4-dichlorophenyl)-1,1-dimethylurea (10(-8) M) magnifies the decreases in oxygen yield and phase lag, it is proposed that the pool which cycles back electrons is in front of the site of 3-(3,4-dichlorophenyl)-1,1-dimethylurea inhibition and is probably the initial electron acceptor pool after Photosystem II.


Sulfide inhibition of photosystem II in cyanobacteria (blue-green algae) and tobacco chloroplasts.

The present study shows that in the presence of 600 nm light, sulfide acts as a specific inhibitor of photosynthetic electron transport between water and Photosystem II in the cyanobacteria Aphanothece halophytica and Synechococcus 6311 as well as in tobacco chloroplasts. In the presence of 600 nm light sulfied affects the fast fluorescence transients as does a low concentration (10 mM) of hydroxylamine; the fluorescence yield decreases in the presence of either chemical and can be restored by the addition of 3-(3,4-dichlorophenyl)-1,1-dimethylurea. In chloroplasts, however, NH2OH, an electron donor at high concentrations (40 mM), relieves the sulfide effect. In the dark, sulfide affects the cyanobacterial fluorescence transients through decrease of oxygen tension. The fluorescence yield increases in a similar pattern to that observed under nitrogen flushing. Upon omission of sulfide in A. halophytica, the characteristic aerobic fluorescence transients return, consistent with the ease of alternation between oxygenic and sulfide-dependent anoxygenic photosynthesis in many cyanobacteria.


Interactions between bacterial membranes and peptidolipids: lysis of micrococcus luteus protoplasts by derivatives of peptidolipidic antibiotics from bacillus subtilis.

The lysis of protoplasts of Micrococcus luteus has been tested with various derivatives of three peptidolipidic antibiotics: iturin A, mycosubtilin and bacillomycin L. The lytic activity is dependent to the nature of the substituting group and to the position of the substituted aminoacid residue. The acetylation of OH groups leads to a decrease of the lytic activity of the natural antibiotics. The methylation of aspartyl residues of bacillomycin L gives a strong lytic activity while natural bacillomycin L has no lytic activity. The methylation of the tyrosyl residue enhances the lytic activities of iturin A and of bacillomycin L-dimethyl ester and reduces that of mycosubtilin. Correlations between the structures of derivatives and their lytic action on M. luteus protoplasts are discussed.


Studies on a membrane-bound and solubilized ribonucleotide reductase preparation from Escherichia coli TAU-.

Ribonucleotide reductase has been shown to be associated with the DNA-membrane complex in Escherichia coli TAU- cells. The membrane-bound enzyme has been released in a soluble form using a combined treatment of 1% sarcosyl (pH 8.0) and 1% sodium deoxycholate (pH 6.5). Allotropic differences in the modulatory effects of ATP, Mg2+, EDTA and dithiothreitol on the membrane-bound and solubilized enzyme activity are discussed.


The interaction of L-ascorbic acid with the active center of myrosinase.

Only L-ascorbic acid activated plant myrosinase (thioglucoside glucohydrolase, EC 3.2.3.1), whereas ascorbic acid analogs did not. The enzyme protein was conformationally changed by the addition of L-ascorbic acid to the spectrophotometric analysis, approx. 1.5 amino residues appeared on the surface of the enzyme and about 2.3 tryptophan residues were buried in the molecule when 1 mM L-ascorbic acid was added. Optimum temperature for the myrosinase activity was approx. 55 degrees C without L-ascorbic acid, but with L-ascorbic acid it was about 35 degrees C; that for beta-glucosidase activity was the same (55 degrees C) with or without L-ascorbic acid. The effect of chemical modification of the functional groups of myrosinase on the interaction of L-ascorbic acid was investigated and the interaction of L-ascorbic acid with the active center of the enzyme is proposed.


Biosynthesis of streptomycin. dTDP-dihydrostreptose synthase from Streptomyces griseus and dTDP-4-keto-L-rhamnose 3,5-epimerase from S. griseus and Escherichia coli Y10.

dTDP-dihydrostreptose synthase from Streptomyces griseus was purfied about 50-fold by removal of protein with polyethyleneimine, (NH4)2SO4 fractionation and gel filtration on Ultrogel AcA44. The synthase preparation was free of dTDP-4-keto-L-rhamnose 3,5-epimerase (dTDP-4-keto-6-deoxy-D-glucose 3,5-epimerase, EC 5.1.3.13) activity. A new enzyme assay using Escherichia coli Y10 as source for the epimerase and dTDP-glucose 4,6-dehydratase (dTDP-glucose 4,6-hydro-lyase, EC 4.2.1.46) was developed. In the presence of excess epimerase the apparent Km for dTDP-4-keto-6-deoxy-D-glucose was determined to be 25 microM. The molecular weight of epimerase and synthase were determined by their elution volumes from a Sephadex G-100 column to be approx. 67,000 and 32,000, respectively. The pH optimum for the epimerase was between 7.5 and 8.5. The intermediate formation of dTDP-4-keto-L-rhamnose in the epimerase reaction could be shown by detection of 6-deoxy-[3H]talose after NaB3H4 reduction. Results which indicate the existence of dTDP-4-keto-6-rhamnose as a free intermediate in the epimerase reaction are reported.


Purification and properties of valyl-tRNA synthetase from Mycobacterium smegmatis.

Valyl-tRNA synthetase from Mycobacterium smegmatis has been purified over 1200-fold by conventional techniques as well as affinity chromatography on valyl-aminohexyl Sepharose columns. The purified preparation is homogeneous by electrophoretic and immunologic criteria. The enzyme is a tetramer of approximate molecular weight of 120,000, composed of a single type of subunit. The synthetase exhibited maximal activity between 35--40 degrees C and pH 6.8--7.0. The pure enzyme though stable for several months below 0 degrees C, loses activity completely at 70 degrees C, for 1 min. The enzyme showed normal Michaelis-Menten kinetic behaviour in the total aminoacylation reaction with Km values of 1.25 microM, 0.1 mM and 1.0 microM for valine, ATP and tRNA, respectively, but the kinetic response deviated from the above pattern in the partial (activation) reaction. Based on these findings, the existence of the enzyme in two molecular forms, modulated by substrate concentration has been suggested; of these, only one may be active in the total reaction, while both forms may function in the phophosphate exchange reaction.


Transfomration of arachidonic acid into 12-hydroxy-5,8,10,14-eicosatetraenoic acid by mouse peritoneal macrophages.

Mouse peritoneal macrophages were incubated at 37 degrees C for 30 min with arachidonic acid (all-cis-5,8,11,14-eicosatetraenoic acid). Oxygenation of arachidonic acid in mouse peritoneal macrophages occurs by two major pathways: fatty acid cyclooxygenase and lipoxygenase. The major metabolite of the latter is 12-hydroxy-5,8,10,14-eicosatetraenoic acid which was identified by gas liquid chromatography on high resolution glass capillary column and mass spectrometry.


Immunohistochemical studies on electron transport proteins associated with cytochromes P-450 in steroidogenic tissues. II. Microsomal NADPH-cytochrome c reductase in the rat adrenal.

NADPH-cytochrome c reductase (NADPH : ferricytochrome oxido-reductase, EC 1.6.2.4), the flavoprotein which mediates the NADPH-dependent reduction of cytochromes P-450 in adrenocortical microsomes, has been localized immunohistochemically at the light microscopic level in rat adrenal glands. Localization was achieved through the use of sheep antiserum produced against purified, trypsin-solubilized rat hepatic microsomal NADPH-cytochrome c reductase in both an unlabeled antibody peroxidase-antiperoxidase technique and an indirect fluorescent antibody method. The sheep antibody to rat hepatic microsomal NADPH-cytochrome c reductase concomitantly inhibited the NADPH-cytochrome c reductase and progesterone 21-hydroxylase activities catalyzed by isolated rat adrenal microsomes. When sections of rat adrenal glands were exposed to the reductase antiserum in both immunohistochemical procedures, positive staining for NADPH-cytochrome c reductase was observed in parenchymal cells of the three cortical zones but not in medullary chromaffin cells. The intensity of staining, however, was found to differ among the three cortical zones, with the most intense staining being found in the zona fasciculata and the least in the zona glomerulosa. The intensity of staining was also found to differ among cells within the zona fasciculata. These immunohistochemical observations demonstrate that microsomal NADPH-cytochrome c reductase is not distributed uniformly throughout the rat adrenal cortex.


Insulin action in isolated fat cells. I. Effects of divalent cations on the stimulation by insulin of glucose uptake.

The effects of divalent cations, in particular Ca2+ and Mg2+, on glucose uptake by rat isolated fat cells in the presence and absence of insulin have been studied. EDTA (disodium salt) was used to deplete the bovine serum albumin present in the incubation medium of endogenous divalent cations prior to incubation with the cells, but was not present in the incubation medium during the incubation of the cells. The removal of Ca2+ and Mg2+ from the incubation medium did not affect the basal glucose uptake, but abolished the ability of insulin to stimulate glucose uptake by the cells. Addition of 25 microM MgCl2 or CaCl2 to the incubation medium restored a significant insulin stimulation, and this stimulation was maximal when 0.1 mM MgCl2 or CaCl2 had been added. SrCl2 and BaCl2 were also effective in restoring the insulin stimulation, but did not substitute fully for Ca2+ and Mg2+ in the incubation medium. Possible explanation for these observations are discussed.


Red cell proteins. I. Two-dimensional mapping of human erythrocyte lysate proteins.

Human erythrocyte lysate proteins were resolved into over 250 discrete spots by two-dimensional electrophoresis using isoelectric focusing in the first dimension and electrophoresis in the presence of sodium dodecyl sulfate, (SDS) in the second. The overwhelming excess of hemoglobin has made such analyses difficult in the past. However, with the ISO-DALT two-dimensional electrophoresis system, large numbers of red cell proteins can be mapped in the presence of hemoglobin. When hemoglobin and several other major proteins are removed by adsorption to DEAE-cellulose, additional minor components are seen, giving a total of over 275. With the use of purified preparations, the map positions of five cell enzymes or their subunits were determined: pyruvate kinase, catalase, glucose-6-phosphate dehydrogenase, hypoxanthine phosphoribosyltransferase, and carbonic anhydrase. The mapping techniques described complement and extend those traditionally used to find human red cell protein variants.


Fractionation of antibodies to L-cell colony-stimulating factor by affinity chromatography.

Purified L-cell colony-stimulating factor (CSF) was coupled to cyanogen-bromide-activated Sepharose and used to selectively fractionate antibodies to this factor. With the use of a simplified two-step washing and elution technique, there was 50%--70% binding of the anti-CSF, with recovery of 60%--100% of the bound  material. Both the native antiserum and purified anti-CSF fractions were inhibitory to murine granulocyte-macrophage colony formation. The purified antibodies contained only IgG and were reduced in protein concentration to 0.1% of the serum IgG values. These fractions should prove useful tools for the study of granulocyte and macrophage differentiation.


Response to therapy in IgG myeloma patients excreting lambda or kappa light chains: CALGB experience.

Sixty-eight previously untreated patients with IgG myeloma who were entered into five protocols of Cancer and Leukemia Group B (CALGB) were studied in order to determine the possible influence of excretion of kappa versus lambda urinary light chains on responses to treatment and survival. All patients in these protocols were included if the serum and urine protein abnormalities were confirmed by one of the two group reference laboratories. Pretreatment characteristics of the two groups of patients did not differ significantly. Of 44 patients with kappa Bence Jones proteinuria, 19 patients (43%) had good responses to treatment, whereas only 3 of 24 patients (13%) with lambda Bence Jones proteinuria had good responses (p = 0.02). Survival for the patients excreting kappa light chains was significantly better than survival for patients excreting lambda chains (median survival 31 versus 12 mo, p = 0.02).


Decomposition of DDT and its analogs by soil microflora.

A search was made for active cultures that break down DDT. More than 600 microorganisms were isolated from soils treated for a long time with DDT and from accumulating cultures with DDT or its analogs as the carbon source and with DDT or its analogs and supplementary carbon sources. Cultures capable of degrading DDT under conditions of cometabolism were found among them. It was shown that the dechlorination of the trichloromethyl group of DDT was accomplished by a large number of microorganisms and occurred primarily under conditions of limited aeration. The elimination of chloride ions from the aromatic rings occurred only in the presence of cosubstrates under aerobic conditions and only with two strains. The strain Nocardia sp. 306x carried out complete degradation of DDT with the formation of DBH, DCBP, and an as yet unidentified entirely dechlorinated product. The most profound decomposition of DDT was produced by a culture of Ps. aeruginosa 640x, which entirely degraded the DDT molecule with the formation of phenylacetic acid.


Acute lymphoblastic leukemia with pre-B-cell characteristics.

Blast cells from 6 of 50 patients with acute lymphoblastic leukemia (ALL) displayed intracytoplasmic mu chains in the absence of detectable light chains and surface immunoglobulins. These cells also  expressed lalike and common ALL antigens. Terminal deoxynucleotidyltransferase was detectable in 2 of 5 cases tested. These blast cells are probably related to early B-cell precursors (pre-B cells). In 4 of 6 cases the disease had a tumoral presentation; the prognostic significance of this new subgroup, which accounts for 20% of patients with non-T non-B ALL, remains to be established.


The prolonged exposure of Japanese quail to carbaryl and its effects on neurochemical and blood chemical parameters.

Whole-brain dopamine levels were significantly elevated in male Japanese quail fed 600 and 1200 ppm carbaryl in the diet for 14 weeks. Whole-brain AChE activity, whole-brain and heart norepinephrine levels, adrenal norepinephrine and epinephrine levels and plasma glucose levels were not altered by carbaryl. No effects were noted in the females.


Whole body electron therapy in mycosis fungoides--a successful translational technique achieved by modification of an established linear accelerator.

A translational technique, adapting an established MEL SL75/10 linear accelerator, which gives a satisfactory physical and clinical 3 MeV whole-body electron treatment in mycosis fungoides is described. Tolerance of the patients proved excellent and 16 out of 21 achieved satisfactory remission of the disease. In spite of delivering significant skin doses it was possible to re-treat two relapsing patients without complications and with clinical success.


An intercomparison between two methods of obtaining percentage depth doses for irregular shaped fields and comparison of each method with experimental data for 60Co and 10 MV X rays.

Measurements of central ray tissue-air ratio (TAR) for 60Co for a large number of irregularly shaped fields typically encountered in the cancer clinic were compared with TAR values either calculated by the Clarkson method using scatter air ratios or obtained from tables of square field data using the area/perimeter approach. Irregular field shapes included the "L", pentagonal, rectangular, upper and lower mantle and split fields. Agreement between both calculational methods and direct measurement is within +/- 2% if careful attention is given to central ray position as affected by the proper use of off-centre ratios in air. In the case of 10 MV X-rays, eight randomly chosen irregular fields again yielded an average agreement between both methods of calculation and experimental data of less than 1% and an agreement of less than 0.5% between the A/P and Clarkson calculations.


Sensitivity of Neisseria gonorrhoeae to penicillin in West Germany.

The sensitivity of gonococci to penicillin in Germany has largely decreased in the last 22 years. No significant decrease has occurred, however, in the last three years, but the results show that a slight tendency to decrease may be persisting.


Preservation of Neisseria gonorrhoeae by the gelatin-disc method.

Studies of Neisseria gonorrhoeae are difficult to perform because of the organism's poor survival in vitro. To solve this problem we tried to preserve the organism by a gelatin-disc method. The rate of survival and changes of variations in some biochemical properties of eight strains of N. gonorrhoeae were followed for three years. These studies proved that preservation was satisfactory with only a 1/10 reduction of the living cells. Another trial showed that the organism survived for over six months after being frozen at -20 degrees C. The colonial types, agglutination against red cells from rabbit and guinea pig, and antibiotic susceptibility to penicillin, chloramphenicol, tetracycline, kanamycin, and streptomycin did not change after three years' preservation.


Role of inferior temporal cortex in interhemispheric transfer.

Neurons in inferior temporal cortex of the rhesus monkey usually have large receptive fields that extend well across the midline into both visual half-fields. The responsiveness of these neurons to stimuli in the ipsilateral visual half-field depends on the splenium and anterior commissure, the same pathways necessary for interhemispheric transfer of visual habits. Since inferior temporal neurons have the same trigger features in both half-fields and are usually binocular, they may be the site of the interhemispheric neural convergence that underlies interhemispheric transfer. If so, bilateral removal of inferior temporal cortex should interfere with interhemispheric transfer even when the commissures are intact. To test this, monkeys were trained on pattern discriminations with one eye and then tested for transfer with the other eye. Five experimental monkeys received bilateral inferior temporal lesions and, to restrict input from each eye to one hemisphere, section of the optic chiasm. Ten controls received either bilateral temporal lesions alone, chiasm section alone or remained unoperated. Only the experimental animals showed impaired transfer. These results suggest that inferior temporal neurons mediate interhemispheric transfer by providing perceptual equivalence for patterns in the left and right visual fields, and, by implication, perhaps also for patterns in different parts of the same field.


Role of the periaqueductal grey in vocal expression of emotion.

In 32 squirrel monkeys (Saimiri sciureus) the role of the periaqueductal grey has been investigated by combined stimulation/lesioning and by neuroanatomical experiments. The results are as follows. Firstly, periaqueductal lesions invading the laterally adjacent tegmentum abolish species-specific calls elicitable by electrical brain stimulation. This holds for stimulation sites rostral as well as caudal to this area. The only vocalizations which survive are phonations of an artificial character which can be evoked from the lateral medulla. Spontaneous vocalizations also seem to be abolished. Secondly, vocalizations elicited from the periaqueductal grey are not affected by bilateral lesions in vocalization-eliciting areas rostral to it, but are abolished by lesions in the dorsolateral pons and ventrolateral medulla. Thirdly, the periaqueductal grey receives direct projections from all vocalization-eliciting areas tested, viz. the precallosal cingulate gyrus, gyrus rectus, medial amygdata, central amygdaloid nucleus/substantia innominata, nucleus striae terminalis, dorsal hypothalamus, midline thalamus, periventricular grey, dorsolateral and ventrolateral midbrain tegmentum. Fourthly, the periaqueductal grey projects directly to the nucleus ambiguus, the site of the laryngeal motoneurones. The course of the main bulk of fibres corresponds to the lesion sites effective in abolishing periaqueductally elicited vocalizations. From these results, it was concluded that the caudal periaqueductal-lateral tegmental area is a necessary relay station for all external and internal stimuli capable of inducing species-specific calls. Its position within the stimulus-response loop seems to be on the output side, immediately above the level of motor-corrdination but below that of stimulus recognition.


Effects of plasma cholesterol lowering agents on hepatobiliary lipid metabolism and cholesterol turnover in the rhesus monkey.

The effects of clofibrate, cholestyramine, and neomycin on hepatobiliary lipid metabolism were studied in adult rhesus monkeys in metabolic steady state with intact but exteriorized enterohepatic circulations. Clofibrate (30 mg/kg, id) had no effect on lipid secretion while cholestyramine (150 mg/kg, id) decreased biliary cholesterol secretion rate from 0.19 +/- 0.03 to 0.13 +/- 0.02 mmol/24 h, p less than 0.05. Neomycin (30 mg/kg, id) decreased bile flow from 216 +/- 10 to 191 +/- 7mL/24 h, p less than 0.05, and tended only to decrease bile salt and phospholipid secretion rates. Cholestyramine decreased cholesterol composition from 1.81 +/- 0.22 to 1.30 +/- 0.22 mol %, p less than 0.05, while clofibrate and neomycin had insignificant effects. Cholestyramine and neomycin decreased bile salt pool size from 1 +/- 0.1 to 0.77 +/- 0.15 and from 1.45 +/- 0.16 to 1.13 +/- 0.21 mmol, p less than 0.05, respectively, while clofibrate had no effect. Bile salt synthetic rate was increased only by cholestyramine, i.e., from 0.63 +/- 0.04 to 1.48 +/- 0.26 mmol/24 h, p less than 0.01. Concomitant cholesterol turnover studies revealed that cholestyramine increased the production rate and excretion of cholesterol in the rapidly miscible cholesterol pool and increased the transfer of cholesterol from slow to rapidly miscible pools. Neomycin, on the other hand, decreased the size of the rapidly miscible pool by decreasing production rate without affecting the size of the slowly miscible pool, while clofibrate had insignificant effects.


Polytetrafluoroethylene grafts for vascular access for hyperalimentation.

A polytetrafluoroethylene (PTFE) graft from the brachial artery to the axillary vein may provide a safe and trouble-free means of vascular access in selected patients requiring long-term intravenous feeding in the hospital setting. This route for the administration of solutions supplying total parenteral nutrition (TPN) should be considered in persons whose requirement for parenteral feeding is likely to extend over many months and in whom there are other sources of bacteremia which can imitate or be confused with infection of a conventional central venous feeding line. In a 52-year-old man an 8-mm PTFE graft was anastomosed end-to-side to the brachial artery and the axillary vein and brought through a subcutaneous tunnel on the medial aspect of the upper arm. Access to it was obtained by intermittent puncture with a no. 21 butterfly needle. Hyperosmolar TPN solutions flowed easily by gravity drip into the shunt. The graft seemed resistant to infection. This form of vascular access appears to exclude parenteral feeding lines, solutions and catheters as sources of sepsis.


Crossmatch difficulties following the prophylactic use of Rh immune globulin.

With increasing demand for platelet transfusion the need to use platelets from Rh-positive persons for Rh-negative individuals often arises. The administration of Rho(D) immune globulin (human) in this situation has been recommended, but may cause serologic difficulties owing to the recipient's passive acquisition of antibodies other than anti-D.


Adjuvant chemotherapy for breast carcinoma: psychosocial implications.

Fifty women receiving adjuvant chemotherapy after surgery for Stage II breast carcinoma were interviewed in an effort to describe the psychosocial effect of the treatment. Perceptions of emotional distress and behavioral disruption were rated in five areas, yielding a rating of overall level of disruption and distress. Results showed that all women experienced adverse changes while receiving adjuvant treatments. Of the 50 women, 88% described a decrease in activities related to the effects of adjuvant chemotherapy; 54% reported an increased financial burden; and 41% claimed that their family and/or sexual life had been adversely affected. Despite these adverse changes, 74% of these patients "would definitely" recommend the treatment to friends in a similar situation. Results from this preliminary study may provide useful information to potential participants in adjuvant trials and to the physicians who conduct such trials.


Long-term intravenous therapy with peripherally inserted silicone elastomer central venous catheters in patients with malignant diseases.

Peripherally inserted central venous silicone elastomer catheters were studied in 81 patients who had malignancy requiring prolonged intravenous therapy. The catheters remained in place from 5 to 171 days, with a median of 30 days. Ninety-one percent of the catheters were unassociated with clinical complications. Six percent of 87 inserted catheters were removed due to peripheral thrombophlebitis. Two patients developed subclavian thrombosis, requiring catheter removal. One patient had catheter-related sepsis with Staphylococcus aureus. Bacteria grew from eleven percent of the cultured catheter tips. Indwelling catheters presence did not appear to influence response to antibiotic therapy. We conclude from this pilot study that long-term central venous access with peripherally inserted silicone elastomer catheters has an acceptably low complication rate in a high-risk patient population.


Malnutrition in children with cancer: incidence and consequence.

Overt malnutrition in children with cancer is seen with surprising frequency: up to 37.5% in a group of patients with disease metastatic to or from bone, and 17.5% in a group of newly diagnosed patients with abdominal or pelvic tumors. It appears more frequent in some cancers. e.g., Ewing's sarcoma, than in others, e.g., osteosarcoma. Criteria for diagnosis of overt malnutrition are applicable to the child with cancer. Such overt malnutrition can be successfully and safely treated with intravenous hyperalimentation (IVH). Marginal malnutrition is a state that can be inferred from clinical behavior, although it cannot be objectively diagnosed as yet. Early data suggest that deterioration to overt malnutrition can be averted through IVH. Such nutritional intervention may increase chemotherapeutic tolerance and improve immune defenses. Since childhood cancer is beginning to frequently show excellent outcome, the association of malnutrition with progressive disease strongly suggests investigation of the role of nutritional support.


Alterations of nutritional status: impact of chemotherapy and radiation therapy.

The nutritional status of a cancer patient may be affected by the tumor, the chemotherapy and/or radiation therapy directed against the tumor, and by complications associated with that therapy. Chemotherapy-radiotherapy is not confined exclusively to malignant cell populations; thus, normal tissues may also be affected by the therapy and may contribute to specific nutritional problems. Impaired nutrition due to anorexia, mucositis, nausea, vomiting, and diarrhea may be dependent upon the specific chemotherapeutic agent, dose, or schedule utilized. Similar side effects from radiation therapy depend upon the dose, fractionation, and volume irradiated. When combined modality treatment is given the nutritional consequences may be magnified. Prospective, randomized clinical trials are underway to investigate the efficacy of nutritional support during chemotherapy-radiotherapy on tolerance to treatment, complications from treatment, and response rates to treatment. Preliminary results demonstrate that the administration of total parenteral nutrition is successful in maintaining weight during radiation therapy and chemotherapy, but that weight loss occurs after discontinuation of nutritional support. Thus, long-term evaluation is mandatory to learn the impact of nutritional support on survival, disease-free survival, and complication rates, as well as on the possible prevention of morbidity associated with aggressive chemotherapy-radiation therapy.


Principles of nutritional therapy.

Provision of adequate nutrition makes a major contribution toward improvement of clinical, biochemical, cellular, and psychologic status of the cancer patient in the face of the disease process and the side effects of various treatments. The principles of nutrition support include the following: 1) Malnutrition induced by cancer and its treatment adversely affects the patient and complicates further treatment of the disease. 2) Malnutrition is not an obligatory response of the host to cancer. 3) A rational nutritional therapeutic program for a patient requires analysis of the factors inducing depletion in that patient. 4) Every patient should have an early and periodic assessment of nutritional status. 5) Nutrition therapy, when indicated, should be instituted early. 6) The application and effectiveness of therapeutic programs must become part of the medical audit and general clinical procedure for inpatients and outpatients. 7) The objectives of nutritional therapy are: a) supportive, b) adjunctive, and c) definitive. 8) Nutritional status, tumor growth and anti-tumor treatment are intimately related. 9) Nutritional therapy has the potential for difficulties as well as benefits. 10) The provision of optimal nutrition care requires a multidisciplinary approach with physicians, nurses, dietitians, and pharmacists working as a team with adequate laboratory facilities and administrative and financial support.


Oral feedings in the cancer patient.

The importance of maintaining the cancer patient's nutritional status is now recognized as a major part of the medical care. It is necessary for the oncology team to be aware of the psychological and physiological factors that interfere with food acceptance so that the correct food can be offered at the right time in the most palatable form. The oral route is the preferred method of feeding, and nutritional supplements, chosen according to the individual patient's needs, are of great value in assuring an adequate oral intake. Diagnostic tests and therapy are frequent causes of disruption of the meal schedule and the dietary service must be flexible in providing the patient an opportunity to make up for missed meals. Taste disturbance, nausea, vomiting and mucositis caused by therapy may necessitate periods of intravenous hyperalimentation. Food aversions due to therapy can frequently be prevented by avoiding new or unusual foods in the hours before chemotherapy or irradiation. Regular nutrition counseling during clinic visits and/or hospitalization permits diet modification for specific therapeutic needs. The ultimate goal is the prevention of wasting and debilitation due to malnutrition in the cancer patient.


Nutrition, cancer, and intravenous hyperalimentation.

In over 1000 cancer patients treated with intravenous hyperalimentation (IVH), tumor growth has not been identified and catheter-related sepsis has been minimal. Studies in rats demonstrated that the host benefits more than the tumor during nutritional repletion, and any stimulation of tumor growth in the rat-tumor model could be manipulated with DNA specific drugs to benefit the host. A study of 65 malnourished cancer patients undergoing oncologic therapy and treated with IVH indicated that much of the immune suppression in these patients was the result of malnutrition coincident with or secondary to oncologic treatment. Conclusions reached in this study were that nutritional repletion resulted in a return of skin test reactivity, proper wound healing in the surgical patient, and possibly an increase in response to chemotherapy. Certainly, the use of IVH allowed specific oncologic therapy to be administered to a group of malnourished patients who otherwise might not have been acceptable candidates for intensive antineoplastic therapy.


Parenteral feeding in the management of children with cancer.

Total parenteral nutrition TPN has been used to treat or prevent malnutrition in 65 children with a variety of solid tumors and leukemia in the past 7 years. TPN was used in 58 patients with gastrointestinal complications of surgery, chemotherapy, or radiation; in 2 patients for preoperative correction of malnutrition; and in 8 who were entered into a prospective study. During TPN, general nutrition and appearance improved in all patients. Weight gain was noted in most. Despite gastrointestinal complications, which usually require the interruption of chemotherapy and radiation, treatment could be continued at full dose in 31 children with nutritional support by TPN. TPN was discontinued in 6 patients when blood cultures became positive. Sepsis was treated successfully by removal of the central venous catheter in all 6 and administration of antibiotics in 3. TPN appears to be safe and effective means of combating malnutrition which may occur with cancer and its therapy.


Potent tumor-initiating activity of the 3,4-dihydrodiol of 7,12-dimethylbenz(a)anthracene in mouse skin.

The abilities of the racemic trans-3,4-, 5,6-, and 8,9-dihydrodiols of 7,12-dimethylbenz(a)anthracene to initiate skin tumors in mice were determined by using a two-stage system of tumorigenesis. The 7,12-dimethylbenz(a)anthracene trans-3,4-dihydrodiol was found to be much more active as a tumor initiator than the parent hydrocarbon. The 7,12-dimethylbenz(a)anthracene trans-5,6- and 8,9-dihydrodiols were essentially inactive as skin tumor initiators. Our results suggest that the 3,4-dihydrodiol of 7,12-dimethylbenz(a)anthracene is a proximal carcinogen and that the "bay region" diol-epoxide may be the ultimate carcinogenic form of DMBA.


Ozonation of mutagenic and carcinogenic polyaromatic amines and polyaromatic hydrocarbons in water.

The Salmonella-microsome assay for mutagenesis was used to determine the effect of ozone on the mutagenesis of selected carcinogens and mutagens in water. Short periods of ozonation were shown to completely inactivate the mutagenicity of several polyaromatic amine mutagens including acriflavine, proflavine, and beta-naphthylamine. Selected polyaromatic hydrocarbons were also sensitive to ozonation. Kinetic studies revealed that the mutagenicity of benzo(a)pyrene, 3-methylcholanthrene, and 7,12-dimethylbenz(a)anthracene was destroyed after short periods of ozonation. To correlate loss of mutagenicity with loss of carcinogenicity, two polyaromatic hydrocarbons were treated with ozone, extracted from water with hexane, and tested for carcinogenicity in mice. When 7,12-dimethyl-benz(a)anthracene and 3-methyl-cholanthrene were treated with ozone, there was a substantial reduction in carcinogenicity compared to control groups treated with oxygen alone. However, a small number of tumors developed in the group of animals receiving a hexane extract of ozonated 7,12-dimethylbenz(a)anthracene. This activity may be due to breakdown products of 7,12-dimethylbenz(a)anthracene that are not mutagenic.


Large animal studies of hyperthermia and irradiation.

Investigators who have studied hyperthermic response of spontaneous animal tumors have reported complete remission for variable periods of 15 to 38% of tumors treated. Normal tissue complications were minimal. Long-term control appears more likely if irradiation is combined with hyperthermia, but information is lacking to date to confirm this. Dose-response assays of radiation alone have been done which would make comparisons with hyperthermia and radiation more meaningful. Probabilities for increasing tumor control without significantly increasing normal tissue response can be estimated better from such assays. Spontaneous tumors in companion animals have advantages over rodent tumor systems of relatively larger treatment fields;  longer follow-up times are possible, and serial monitoring of a variety of clinical data can be done. Experience of investigators using these tumors has shown that animal owners and referring veterinarians are most cooperative in reasonable, humane approaches to experimental cancer therapy.


Sister chromatid exchange induction resulting from systemic, topical, and systemic-topical presentations of carcinogens.

Chinese hamster cheek pouch mucosal cells were examined for in vivo sister chromatid exchange formation resulting from the exposure of animals to carcinogens presented in three manners: systemic, topical, and systemic-topical combination. The systemic presentation of cyclophosphamide (5 or 10 mg/kg) through i.p. injection resulted in an increase in sister chromatid exchanges from 4.8 to 9.9 per cell. Topical application of 7,12-dimethylbenz(a)anthracene (0.5% in mineral oil, 0.1 ml) resulted in an increase in the sister chromatid exchange frequency to 11.5/cell, as compared with a value of 5.0/cell in animals treated only with mineral oil. Systemic administration of 8-methoxypsoralen (0.5, 1, 2.5, or 5 mg/kg) through i.p. injection, followed by activation with topical near-ultraviolet light (3.75 x 10(4) ergs/sq mm at 365 nm) resulted in an increase in sister chromatid exchange, reaching 15.4/cell at 5 mg 8-methoxypsoralen per kg. Exposure of animals to 8-methoxypsoralen or near-ultraviolet light alone, but not in combination, did not produce an increase in sister chromatid exchange. Sister chromatid exchange frequencies in cheek pouch cells were also compared with sister chromatid exchange frequencies in marrow cells of identically treated animals to assess the importance of exposure mode and tissue specificity in sister chromatid exchange formation.


Overlapping pathways for repair of damage from ultraviolet light and chemical carcinogens in human fibroblasts.

DNA excision repair was measured in cultured human fibroblasts after single or dual treatments with ultraviolet radiation, 4-nitroquinoline 1-oxide, or N-acetoxy-2-acetylaminofluorene. Three approaches were used to monitor repair: unscheduled DNA synthesis, measured by autoradiography; repair replication, measured by the incorporation of a density-labeled DNA precursor into repaired regions; and excision of ultraviolet endonuclease-sensitive sites. When a single repair- saturating dose of one of the three carcinogens was administered, little stimulation of unscheduled DNA synthesis or repair replication could be observed by additional treatment with one of the other carcinogens. In no instance was total additivity of repair observed. These observations were confirmed by showing that the excision of endonuclease-sensitive sites produced by ultraviolet damage (i.e., pyrimidine dimers) was inhibited by exposure to 4-nitroquinoline 1-oxide and N-acetoxy-2-acetylaminofluorene. The data indicate that the repair of lesions induced by these substances may have common rate-limiting steps, a conclusion previously indicated by the repair deficiency in xeroderma pigmentosum cells in which a single mutation eliminates the repair of damage caused by each of these agents.


Effects of structural changes on the tumor-promoting activity of phorbol myristate acetate on mouse skin.

4a alpha-Phorbol-9,9a-didecanoate, 4a alpha-phorbol-9-myristate-9a-acetate, and phorbol-9-myristate-9a-acetate-3-aldehyde were tested for skin tumor-promoting activity by using 7,12-dimethylbenz(a)anthracene as the initiating agent. There were 30 female ICR/Ha mice/group, and tests were continued for 434 to 461 days. 4a alpha-Phorbol-9,9a-didecanoate and 4a alpha-phorbol-9-myristate-9a-acetate were devoid of tumor-promoting activity. Phorbol-9-myristate-9a-acetate-3-aldehyde resulted in 10 mice with papillomas, 2 of which also bore squamous carcinomas of the skin. The positive control group, in which phorbol myristate acetate was used as promoting agent, resulted in 30 mice bearing multiple papillomas and 15 bearing squamous carcinomas of the skin. The effects of structural and stereochemical changes on tumor-promoting activity suggest that a primary interaction of the phorbol ester series is binding at specific sites on the plasma membrane.


Variable oral absorption of hexamethylmelamine in man.

Plasma levels of hexamethylmelamine were measured by a sensitive and specific gas chromatographic assay in 11 patients after they received oral doses ranging from 120 to 300 mg/m2. Peak levels were reached in 0.5--3 hours and ranged from 0.2 to 20.8 micrograms/ml. The area under the curve ranged from 70.2 to 3606.6 micrograms/ml x minute. The half-life of the beta-phase of elimination also showed considerable variability, ranging between 4.66 and 10.2 hours. These parameters were not related to the dose administered. No plasma accumulation of the drug was observed after daily treatment with hexamethylmelamine for 14--21 days.


Disposition of 5-methyltetrahydrohomofolate in mice, dogs, and monkeys.

The pharmacologic disposition of [methyl-14C]5-methyltetrahydrohomofolate (MTHHF) has been studied in mice, dogs, and monkeys after parenteral administration of doses of 150 and 1500 mg/m2. Thin-layer chromatography and high-pressure liquid chromatography were used to separate the parent compound from its products. Following ip injection of MTHHF into mice, serum levels rose to a maximum within 1 hour and decreased with a half-life of about 50 minutes. The liver, kidneys, and small intestine of mice contained concentrations of MTHHF greater than that of serum. For dogs and monkeys given an iv injection of MTHHF, serum levels declined in three phases. The third phase was probably associated primarily with loss of the drug from the liver and intestine, where relatively large amounts were retained. Within 24 hours after administration of MTHHF, the percent recovery in the urine was 69% for mice, 78% for dogs, and 50%--60% for monkeys. For mice and dogs an additional 2%--17% was recovered in the feces within 24 hours. Over a period of 3 days after injection with the high dose in a monkey, 24% of the dose was present in the feces. For all three species, 90%--99% of the radioactivity in the serum, urine, and feces was unchanged MTHHF, indicating that very little metabolism or decomposition had occurred. Less than 0.3% of the dose was recovered from monkeys as radioactive CO2.


The Princess Margaret Hospital study of ovarian cancer: stages I, II, and asymptomatic III presentations.

An analysis of 231 patients with stages I, II, and asymptomatic III ovarian cancer, studied in a prospective randomized-stratified trial, is presented. None of the stage IA patients with well-differentiated tumors have had disease relapses; one fourth of the patients with poorly differentiated tumors have had disease relapses throughout the peritoneal cavity. There is therefore little justification for pelvic radiation alone as postoperative therapy for stage IA ovarian carcinoma. For stage IB, II, and asymptomatic III presentations, patients with an incomplete initial pelvic operation had poor survival characteristics with all tested therapies. For patients in whom the operation was completed, abdominopelvic radiation was superior to pelvic radiation alone or followed by chlorambucil, with respect to long-term survival and control of abdominal disease. The effectiveness of abdominopelvic radiation was independent of stage or histology. The value of abdominopelvic radiation was most strikingly seen in patients with no visible residual tumor.


Histologic grade in advanced ovarian cancer.

The histologic grade of epithelial ovarian tumors was found to be a major prognostic factor for survival in patients with advanced (stage III-IV) disease. In addition, a grading system based on cytologic detail (modified Broders' grades I-IV) identified groups with a differential response to chemotherapy. The overall improvement in survival observed with combination chemotherapy was related primarily to an increased survival of patients with grade II and III lesions. Although the survival of patients with grade I lesions was markedly better than for patients with grade IV lesions, in neither case was it influenced by the choice of single-agent or combination chemotherapy. In prospective clinical trials in advanced disease, modified Broders' grades should be included as a separate stratification factor.


Combination chemotherapy for alkylator-resistant ovarian carcinoma: a preliminary report of a Southwest Oncology Group trial.

The Southwest Oncology Group has carried out a phase III study of combination chemotherapy in patients with advanced epithelial ovarian cancer who had become resistant to prior drug treatment. Patients previously treated with only alkylating agents were given adriamycin-5-fluorouracil-hexamethylmelamine-cis-dichlorodiammineplatinum(II) combination. There have been 12 partial and two complete responses among 29 evaluable patients (at least one full treatment course) entered in this four-drug combination, for a 48% response rate. Two (7%) additional patients showed objective improvement. The median duration of response was 5.8 months. Ten (34%) of 29 patients have died. The median duration of survival was 12+ months. Patients who had previously received adriamycin plus alkylating agent therapy were treated with 5-fluorouracil-hexamethylmelamine-cis-dichlorodiammineplatinum(II). Only 23 (31%) of these 74 patients have had partial responses while an additional 7% have had objective "improvement"; 27 (36%) of these patients have died. The median duration of survival was 14 months. Both drug regimens were well-tolerated. There were no drug-related deaths. Although thrombocytopenia was dose-limiting in up to 40% of the patients, the mean lowest platelet counts in the four- and three-drug regimens were 116,000 and 123,000/mm3, respectively. There were no cases of serious azotemia (ie, BUN level greater than 40 mg/100 ml) or severe peripheral neuropathy.


Phase II trial of cyclophosphamide, hexamethylmelamine, adriamycin, and cis-dichlorodiammineplatinum(II) combination chemotherapy in advanced ovarian carcinoma.

A monthly four-drug regimen of cyclophosphamide, adriamycin, and cis-dichlorodiammineplatinum(II), each given iv of Day 1, and hexamethylmelamine, given orally on Days 1--14 (CHAP), was administered to 39 women with advanced epithelial ovarian carcinoma who had previously failed alkylating agent therapy. Of 35 evaluable patients with a measurable disease parameter, seven (20%) achieved a clinical complete response and ten (29%) achieved a clinical partial response. The median duration of complete response is greater than 9 months and the median duration of partial response is 4 months. Ninety percent of the patients required dose adjustments because of profound leukopenia, thrombocytopenia, or gastrointestinal intolerance. CHAP is an active but toxic regimen in the management of advanced ovarian cancer. The toxicity encountered reflected the intensity of the drug schedule as well as the combined influences of advanced stage of disease, poor nutritional and performance status, and prior therapy. The efficacy of this CHAP regimen warrants a controlled trial compared to other active drug programs.


The CHAD and HAD regimens in advanced ovarian cancer: combination chemotherapy including cyclophosphamide, hexamethylmelamine, adriamycin, and cis-dichlorodiammineplatinum(II).

Combination chemotherapy consisting of hexamethylmelamine, adriamycin, and cis-dichlorodiammineplatinum(II) was given to 27 patients with advanced epithelial ovarian cancer whose disease had progressed while receiving therapy including alkylating agents. Eighteen patients (67%) had objective partial or complete remissions for a projected median duration of 7 months. Based on this high level of activity in this poor-risk group, the three drugs were combined with cyclophosphamide in a four-drug regimen (CHAD) for initial therapy suitable for outpatient administration. Nineteen of 21 evaluable patients (90.4%) receiving CHAD responded. Complete clinical remissions were noted in ten patients (47.6%) and were confirmed at "second-look" laparotomy in two of these. An additional five patients without evaluable disease after initial surgery are free from relapse at 4--15 months. Toxicity was only moderately severe; 28% of the patients had wbc counts less than 2000/microliters at some point. Nephrotoxicity from cis-dichlorodiammineplatinum(II) was almost completely prevented by a 2-hour outpatient regimen of hydration and diuresis. A prospective randomized trial is planned comparing CHAD to melphalan in previously untreated women with advanced ovarian cancer.


Once weekly total-skin electron-beam therapy for mycosis fungoides: 7 years' experience.

A total of 115 patients with mycosis fungoides were given total-skin electron-beam therapy (TSEB), utilizing 3.5-mev electrons at doses of 400 rads to the entire skin surface once a week for 6--8 consecutive weeks. Prompt relief of symptoms and regression of lesions were observed in all patients. Of the 81 patients at risk for 12--91 months (median, 24 months) following TSEB, initial unmaintained remission lasted 6--69 months (median, 19 months). No untoward immediate or late effects have been noted in the bone marrow or normal skin which was irradiated. The duration of remission following TSEB correlated well with lymphocyte responsiveness to various mitogens and antigens, but not with the initial response. Thymic hormone factor levels (Facteur Thymic Serique) were elevated in the majority of these patients with mycosis fungoides.


Mycosis fungoides: electron beam therapy in England.

Since 1962, total-skin electron-beam therapy has been available in London, England, for the treatment of patients with mycosis fungoides and skin lymphomas. The 6-mev linear accelerator produces a pencil beam of electrons at 7 mev, which are scattered through a brass scatterer and decelerated by either of two carbon decelerators or a copper decelerator to produce a beam of effective energy of 2.5, 3, or 3.5 mev; these beams have an 80% isodose distribution at 5.5, 8, and 11.5 mm, respectively. The patient receives 200 rads to the anterior, posterior, and right and left lateral fields, for a total dose of between 1600 and 2600 ras in ten treatments over 5--7 weeks. Complete clearing of the disease can be predicted in all patients except those with the most advanced tumors. However, the duration of remission after electron-beam therapy is approximately 18 months and we are presently investigating the combination of psoralens and ultraviolet light therapy as maintenance treatment following lower-dose electron-beam therapy.


Combined therapy for patients with mycosis fungoides.

In a comprehensive mycosis fungoides program, 60 patients have been seen with a pathologic diagnosis of this disease. Forty-four patients with advanced disease were referred for radiation therapy. Three treatment techniques were identified in which 14 patients were treated with localized fields using electrons or whole-body electron-beam therapy with doses of less than 3000 rads, 21 patients were treated using the Stanford technique with tissue doses of between 3000 and 4000 rads, and nine patients were treated with six cycles of mechlorethamine, vincristine, prednisone, and procarbazine or cyclophosphamide, vincristine, prednisone, and procarbazine following the electron-beam therapy. The actuarial survival rate was 45% at 1 year for the 14 patients with localized electron-beam therapy, whereas  the actuarial survival rates were 83% for patients treated with whole-body electron-beam therapy and 100% for patients treated with whole-body electron-beam therapy followed by four-drug chemotherapy. The recurrence-free interval for these three groups correlates with these observations. A central nervous system recurrence has been observed in the combined-therapy group.


A 10-year experience with topical mechlorethamine for mycosis fungoides: comparison with patients treated by total-skin electron-beam radiation therapy.

A group of 243 patients with mycosis fungoides (MF) received treatment with topical applications of dilute aqueous solutions of mechlorethamine and/or systemic chemotherapy over the past 10 years. The likelihood of a complete and relapse-free remission and survival was found to correlate inversely to the magnitude of disease as denoted by a simple staging system. Although disease-free intervals of greater than 3 years have occurred thus far in 32 (13%) patients, the permanency of these remissions and the curability of disease remain uncertain because of the variability of disease progression characteristic of MF. Comparison of treatment results with those published on a large group of patients treated with total-skin electron-beam radiation therapy indicates that the chemotherapeutic approach to the treatment of MF is equally effective in promoting survival.


Electron-beam therapy for mycosis fungoides: the Stanford University experience.

The use of high-dose electron-beam therapy for mycosis fungoides at Stanford University is reviewed. Since 1966, 140 patients have been treated in this fashion. Their clinical characteristics including initial extent of disease are reviewed. The results of routine staging studies are examined. Eighty-four percent of the patients achieved an initial complete remission; this was inversely related to the initial extent of skin involvement. The overall survival rate was 46% at 10 years with the major prognostic factor being the initial extent of skin involvement. Other factors which had an influence on the patient's progrnosis included the presence of palpable adenopathy, the patient's age, the achieving of an initial complete remission, the initial dose of electron-beam therapy, and the treatment with adjuvant topical mechlorethamine. The development of a rationale for the treatment of patients with all stages of mycosis fungoides based upon these clincal observations is discussed.


[Desmodonte renewal in baboons (Papio papio)].

Turnover of desmodontal ligament is analysed in the Baboon using labeling, biochemical and histological techniques. In Baboons, as previously observed in rodents, turnover of proteins labelled by 3H glycine may be described by a single exponential after the ninth day, the half life of which is more than two weeks. Biochemical data show that proteins renewed are mostly collagen.


[Visually guided movement in the cat: difference in the effects of a bilateral lesion of the thalamic nucleus ventralis lateralis performed either before or after training].

Bilatéral lesions of the thalamic nucleus ventralis lateralis were performed in cats, either after [1] or before [2] training to reach a moving target with the forepaw. In situation [1], lesions were ineffective; in situation [2] the animal never reached normal scores. VL thus seems to be essential only in the acquisition phase of the tested visuomotor performance.


[Interaction of glucocorticosteroids with glucagon in the control of argininosuccinase activity in the fetal rat liver].

Exogenous glucagon, but not glucocorticosteroids cause a rise in argininosuccinase activity in the liver of 21.5 day old fetuses. When fetuses are deprived of glucocorticosteroids, glucagon is unable to promote the enzymic activity. This study indicates that glucocorticosteroids are required for the induction of the argininosuccinase activity by glucagon.


[Evidence of 5 alpha-androstane-3 beta, 6 alpha, 17 beta-triol and of 5 alpha-androstane-3 beta, 7 alpha, 17 beta-triol in the anterior pituitary of the prepuberal male rate].

Incubation of 3H-5 alpha-androstane-3 beta, 17 beta-diol with anterior pituitaries from immature male Rats led to the major production of polar radio-metabolites. Of these, 53 and 27% were identified with 5 alpha-androstane-3 beta, 6 alpha, 17 beta-triol and 5 alpha-androstane-3 beta, 7 alpha, 17 beta-triol, respectively. The formation of these trihydroxysteroids may be involved in the intracellular regulation of 5 alpha-androstane-3 beta, 17 beta-diol levels.


[Is the process of localized lysosomal exocytosis responsible for the cytolytic action of killer T-lymphocytes?].

In this paper, we formulate the hypothesis that in the process of target cell lysis a lysosomal enzyme regurgitation, performed by killer cells at the level of the target effector junction, accounts for the target lesion which precedes the lysis (lethal hit). This process of exocytosis, similar to the one described previously in polymorphonuclear neutrophils is supported by cytological studies performed directly on identified killers isolated by micromanipulation. Light and electron microscopy observations confirm a previous report which describes the effector cells rich in lysosomal bodies. In addition, when a killer cell is associated with a target cell to form a conjugate, lysosomes are concentrated near the cell junction and, after incubation at 37 degrees C, acid phosphatases may be detected at the junction. Lysosomal enzyme exocytosis explains why target lysis needs an effector target binding to occur and also the other conditions required for any exocytosis process such as Ca++ in the medium, integrity of the microtubular apparatus, a low level of cyclic AMP and energy dependancy.


[Effects of C1-C8 n-alcohols on the growth of 3T3 mouse fibroblasts].

The effect of C1-C8 n-alcohols on 3T3 cell growth is studied using flow cytofluorometry. Methanol and ethanol markedly lengthen either the duration of G1, or that of G2+ M when present in relatively higher doses. The effect of longer chains is always to increase G2+M significantly. This may be due to deviations depending on alcoholic chain length in membrane lipid fluidity.


[Evidence of abnormalities in net sodium and potassium fluxes in erythrocytes of patients with essential hypertension].

A new and simple laboratory test for measuring net Na+ and K+ fluxes in Na+-loaded/K+-depleted human erythrocytes was developed and applied to hypertension. Moderate essential hypertension was characterized by a constant increase in net K+ influx; more severe cases showed a drop in net Na+ efflux. Na+ and K+ erythrocyte fluxes were found to be normal in hypertension of renal origin.


Hypertension following denervation of aortic baroreceptors in unanesthetized dogs.

After cervical aortic nerve section, mean arterial pressure in the unanesthetized dog increased by an average of 7.4 mm Hg. Following a more extensive denervation of aortic arch receptors by section of intrathoracic vagal branches, arterial pressure increased by 16.7 mm Hg. The above changes were seen in the stable state after the effects of surgery had disappeared. In both cases carotid baroreceptors were functional. After administration of nitroglycerin and phenylephrine subsequent to either denervation procedure, blood pressure changes were larger and heart rate responses were smaller than in the control state. The unanesthetized dog regulates mean arterial pressure at a higher-than-normal pressure after aortic baroreceptor denervation. Reflexes from the aortic baroreceptors continuously participate in the normal control of mean arterial pressure. Section of the cervical aortic nerves only partially denervates aortic baroreceptors. Our findings may be relevant to human essential hypertension.


Neither dorsal root nor baroreceptor afferents are necessary for eliciting the renal responses to acute intravascular volume expansion in the primate Macaca fascicularis.

We determined the contribution of the dorsal roots, vagi, and sino-aortic nerves to the renal responses to acute isotonic, isooncotic intravascular volume expansion in the nonhuman primate, Macaca fascicularis. Expansion of the estimated blood volume by 15% produced a significant natriuresis and diuresis. There was no significant difference between the time to peak response for either. Neither dorsal rhizotomy (C6-T7) nor vagotomy and sino-aortic denervation had a significant effect on these responses. We conclude that these pathways are not necessary for eliciting the renal responses to hypervolemia in the nonhuman primate.


Specific antibody to hog renal renin and its application to the direct radioimmunoassay of renin in various organs.

We produced anti-hog renin antibodies using as antigens pure hog renal renin that either had been insolubilized or conjugated to tetanus toxoid. High titer antibodies were obtained, which demonstrated different cross-reactivity with renins from other species. A direct radioimmunoassay for renin was developed using antibody, monoiodinated 125I-hog renin, and various methods for separating free and antibody-bound trace. This assay was capable of detecting 40 pg of hog renin and was applied to the determination of renin in hog blood and other organs. Based on the direct measurement of renin by this radioimmunoassay, the renin-like activity (i.e., the ability to generate angiotensin I from renin substrate preparations) of the pituitary gland was found to be due mostly to true renin, whereas the renin activity of other hog tissues, including the adrenal gland, liver, lung, spleen, and submaxillary gland, was not identified as renin and may have been due to cathepsins.


Effect of acute ischemia, nitroglycerin and nitroprusside on regional myocardial thickening, stress and perfusion. Experimental echocardiographic studies.

The purposes of this study were to demonstrate that echocardiography can be used to demonstrate the systolic wall thinning of acutely ischemic myocardium, and to compare the effects of nitroglycerin and nitroprusside on systolic thinning, wall stress and perfusion of ischemic myocardium. In 37 dogs, the ratio of end-systolic-to-end-diastolic posterior wall thickness fell from 1.30 +/- 0.02 to 0.88 +/- 0.01 ((p less than 0.001) after circumflex coronary occlusion; perfusion of the area supplied by the occluded artery fell from 98.2 +/- 7.5 ml/100 g/min to 36.5 +/- 2.9 ml/100 g/min (p less than 0.001). Nitroglycerin and nitroprusside were given to lower mean arterial pressure by 7% and 15%. Despite the reduction in coronary perfusion pressure, transmural perfusion, endocardial/epicardial perfusion ratio and systolic thinning remained constant. Both drugs reduced the ischemic "wall stress index" (ventricular pressure x ventricular diameter/wall thickness) by almost 50%. Thus, both nitroglycerin and nitroprusside were equally beneficial in this model of acute myocardial ischemia.


IgE and IgG4 subclass in atopic families.

Raised levels of IgG4 were present in twelve (35%) of thirty-four asthmatic children and raised levels of IgE in twenty-three (68%). Eighteen of 104 first degree relatives also had raised levels of IgG4, thirteen had no history of atopic disease and nine failed to give positive skin reactions to Dermatophagoides pteronyssinus or to mixed grass pollens. Fifteen relatives had raised IgE, five without symptoms. No relationship was noted between either raised levels of IgE or IgG4 and infant feeding. Although these immunoglobulin patterns were not consistently associated with symptoms they did tend to be associated in atopic families and it is suggested that the same polygenic factors may govern the IgE and IgG4 levels.


Gas chromatographic determination of glibenclamide in plasma.

A gas-chromatographic method for glibenclamide determination in plasma is described. It involves derivatization of the drug with dinitrofluorobenzene and the use of an electron-capture detector. The quantitative evaluation is performed using tolbutamide as internal standard. Characteristics and specificity of the method for the principal metabolite of glibenclamide are examined.


Solid-phase radioimmunoassay for norethisterone.

Exposure of polystyrene tubes to solutions of glutaraldehyde was shown to provide a stable and consistent surface for immobilising an unpurified norethisterone-specific antiserum. Properties of the solid-phase antiserum were evaluated with respect to its application in developing a solid-phase radioimmunoassay for norethisterone. The assay proved to be sufficiently accurate, precise and sensitive for routine use. The technology is considered to be suited to automation.


Urinary excretion of orotic acid, orotidine and other pyrimidines in a patient with purine nucleoside phosphorylase deficiency.

Urinary orotidine and orotic acid have been determined in a patient with purine nucleoside phosphorylase (PNP) deficiency under various dietary therapeutic conditions. For this purpose a new procedure for the analysis of both compounds has been developed, consisting of prefractionation with Dowex 1X8, followed by two HPLC steps on a micro Bondapak NH2 and a micro Bondapak C18 column. With this method normal as well as slightly elevated excretions of orotic acid have been found in our patient. No evidence was obtained for inhibition of OPRT by purine (deoxy)nucleosides as a cause of pyrimidine starvation. A significant increase of urinary orotidine was found after loading with allopurinol. For comparison excretory values in a patient with ornithine transcarbamylase deficiency and also in a patient with orotic aciduria type I are shown. The possible cause of the slight increase in urinary orotic acid in our patient has been discussed.


Progressive myoclonus epilepsy: genetic and nosological aspects with special reference to 107 Finnish patients.

In 107 Finnish patients with progressive myoclonus epilepsy (PME), belonging to 74 families, autosomal recessive inheritance was evident. The sex ratio was 48:51, the corrected proportion of affected sibs being 0.260. Of 68 marriages 15, or 22%, were consanguineous; several of the parents were related and the geographical distribution was of the uneven type typical of young, isolated populations in Finland. The incidence in Finland was estimated to exceed 1:20,000. The clinical picture in the Finnish PME patients was uniform, being identical with that of Unverricht's and Lundborg's patients, but clearly distinct from Lafora disease. The following classification of PME is proposed: (1) PME, Lafora type: onset of grand mal attacks and/or myoclonus around the 15th year of life; rapid and severe mental deterioration, often with psychotic symptoms; short survival; histological finding of Lafora bodies; autosomal recessive inheritance. (2) PME, Unverricht-Lundborg type: onset around the 10th year of life; severity variable, progressive invalidity from myoclonic features associated with mild mental symptoms, time of survival variable, "degenerative" histological changes; autosomal recessive inheritance. (3) Autosomal dominant or otherwise atypical cases of PME. The importance of accurate diagnosis is stressed.


Bacterial endocarditis due to penicillin-resistant Streptococcus viridans.

Bacterial endocarditis remains a formidable diagnostic and therapeutic problem for clinicians. Streptococcus viridans still accounts for 45 to 50 per cent of all cases and between 5 to 10 per cent of all clinical isolates of Streptococcus viridans from patients with bacterial endocarditis may be relatively resistant to penicillin. The case of a 9-year-old child with Tetralogy of Fallot and a Waterston shunt who subsequently developed bacterial endocarditis due to penicillin-resistant Streptococcus viridans following failure of oral penicillin dental prophylaxis is presented. In the face of penicillin resistance, additional considerations for workup, including microbiological assays for antimicrobial synergism become necessary in the selection of a therapeutic regimen.


Therapy for septic shock.

To adequately appraoch the therapeutics of such a diffuse disorder as sepsis, a firm grasp of the multiple pathophysiologic subsets is imperative. With this as a basis, therapy comes as close to applied physiology as is possible in medicine today.


Management of diabetic ketoacidosis.

Many aspects of DKA treatment have become dogma without benefit of critical randomized prospective studies. Although recommendations for low-dose insulin therapy have passed the test of working in practice, many facets of treatment such as bicarbonate vs. no bicarbonate, phosphate replenishment, the merits of rapid vs. slow correction of hyperglycemia and acidosis need to be prospectively studied. In the final analysis, while insulin, fluid, and electrolyte administration are all important in the treatment of DKA, success most often hinges on careful hour-by-hour monitoring of the patient by a concerned and knowledgeable physician.


Principal sulcus and posterior parieto-occipital cortex lesions in the monkey.

In order to assess the roles of the posterior parieto-occipital and principal sulcus cortices in processing spatial information, both with and without delays, monkeys were given lesions of one or the other area or no lesion and tested on a series of conditional discrimination tasks. There were six basic tasks, and the cue locations differed from the response locations within each task. The first two had mixed dimensions in that the location of the cue indicated the color of the correct response alternative or the color of the cue indicated the correct response location. The animals were trained preoperatively on these tasks and then tested for postoperative retention. For the next two tasks color was the only relevant dimension, but different colors were used for the cues and response alternatives. For the final two tasks location was the only relevant dimension. When an animal learned a given task it was subsequently tested on it with a 5-sec. delay. The deficits observed suggest that the roles of these areas can not be expressed simply in terms of spatial or delay functions. No group appeared to be impaired on any delay task on which it was tested. The animals with the posterior cortex removed had a great deal of difficulty in using information from one dimension to identify the correct response alternative by the other dimension, but not when the relevant cue and response dimensions were the same. These results suggest that the posterior parieto-occipital area is involved in making associations across dimensions with in the visual modality. The animals with the principal sulcus removed appeared to have difficulty only when they were required to respond with reference to the colors of the response alternatives. On such tasks the relevant dimension (color) was not available until the presentation of the response array. On the other tasks the correct response location could be determined from the cue presentation alone, that is, before the presentation of the response array. This suggests that these animals had a tendency to respond without reference to the color dimension, but rather just to the presentation of the response array, and therefore a principal sulcus function of inhibiting a response until the appropriate information has been processed.


Urinary enzyme assays in toxicological studies in the rat and marmoset.

The relative merits of the automated, fluorimetric assay of urinary enzymes and cell exfoliation were compared with other commonly used tests for renal damage. Two types of nephrotoxic agent were used, causing crystal nephropathy and acute tubular necrosis respectively. Groups of marmosets were given one of two drugs known to cause crystal nephropathy. One agent caused intermittent increases in urinary enzyme excretion and an early increase in cell excretion which was not sustained. The second agent in contrast caused elevated cell and enzyme excretion, increasing throughout the period of administration. A nephrotoxic anti-tumour agent also caused increases in cell and enzyme excretion when given to marmosets. The early changes produced by this agent were studied using catheterised rats. Hourly samples of urine were collected and urinary beta-glycosidase excretion was found to give an early indication of renal damage, which correlated with albuminuria and glycosuria. The fluorimetric assay of urinary enzymes provides a sensitive, non-invasive test of nephrotoxicity.


Isolation and renal localisation of urokinase.

Physico-chemical characteristics of urokinase in urine were studied by immunological and chemical methods. By agar zone electrophoresis, commercial urokinase preparations could be separated into an anodic and cathodic fraction. The latter reacted with urokinase antibodies with two precipitation bands. Band I displayed the major part of urokinase activity and migrated as a beta-globulin with a molecular weight of 32,000 daltons. Band II showed immunological identity with human serum, human albumin, alpha-2-macroglobulin and alpha-2-HS-glycoprotein. The specific activity of the cathodic fractions was up to 80,000 ploug units/mg protein. The ratio esterase/fibrinolytic activity did not change during the purification procedure. Further purification of the fractions with higher specific activity by affinity chromatography was unable to eliminate material cross reacting with human antisera (Band II). These findings permit the conclusion, that urokinase activity in urine is not confined to a homogeneous protein fraction. Activity is found both in a low molecular weight fraction and in a high molecular weight complex which contains serum proteins. These cannot be removed by exhaustive purification procedures and may play an important role in stabilizing and/or protecting urinary urokinase against proteolytic degradation. With Todd's technique diffuse fibrinolytic activity could be demonstrated in the kidney in the iuxtamedullary border region, (venae arcuatae, venae interlobulares, vasa recta) and in the epithelium of the calyces. Urokinase activity was specifically blocked by highly purified urokinase antibodies and could thus be distinguished from nonspecific proteolytic activity. The topographic relationship to medulla and uroepithelium may point to a role of urokinase in maintaining patency in slow flow systems.


Cellular control of growth in cultures of Tetrahymena. 3. The influence of Fe-medium and aeration on cellular ultrastructure.

Cultures of Tetrahymena pyriformis were grown with and without the addition of Fe and with no aeration. The same cultures were used both for determinations of the cellular Fe and Ca concentrations and the exchange of Ca (reported earlier), and for the ultrastructural study. In all cultures there was an increase in rounded, tubuli-deficient mitochondria at the transition to the prestationary growth phase. In the non-aerated culture (high cellular Fe content) these changes were less marked. In the Fe-deficient culture, however, these mitochondrial changes were seen as early as the late exponential growth phase, and tubuli-degeneration then increased during the prestationary growth phase. In this culture an irregular infolding of the outer mitochondrial membranes occurred. These changes are discussed in correlation with cytochromes, Fe-dependent desaturation of fatty acids, and high Ca concentration (non-aerated cells). During the prestationary growth phase of the non-aerated culture there was a marked increase in the amount of mitochondrial tubuli. In the organelles identified as peroxisomes there was, in all the cultures, an increased granular density of the matrix at the transition to the prestationary growth phase (in the Fe-deficient cells this occurred in the late exponential growth phase). This was correlated with an increased peroxisomal activity. The Fe-deficient culture has cells with very irregularly formed peroxisomes. This organelle was in all the cell-material very sensitive to the method of fixation. In the Fe-deficient late exponential cells there are long, bifacial pieces of RER (one side rough and the other smooth) which later undergo degradation. Many lipid droplets were seen at the ends of RER. Structures which in the literature have been called 'ergoplasm-like stacks of flattened rough cisternae' were found in the non-aerated exponential cells. They were absent from prestationary cells. In all the cultures there was an increased aggregation of the ribosomes in the cytoplasm during the prestationary growth phase. This was correlated with the accumulation of Ca in this cell fraction. An explanation is suggested regarding the earlier reported variations in the exchange of Ca, found in all the types of cultures at the transition to the prestationary growth phase.


Retinal hemorrhage in the battered child.

The presence of retinal hemorrhage in head-injured children under 3 years of age is believed to be pathognomonic of battering. When a group of battered children was compared to head-injured children due to other causes, the high incidence of retinal hemorrhage in the battered children was contrasted with the absence of retinal hemorrhage produced by other causes of head injury.


[Causes and risks in a interruption of systemic immunosuppressive therapy following kidney transplantation].

The immunosuppressive therapy of 22 transplant recipients was temporarily suspended 25 times resulting in rejection reactions in 7 cases with the eventual loss of five transplants. One patient succumbed to fungal infection following the loss of kidney function. No connection between the risk of rejection and the previous number of rejection treatments or the histocompatibility could be established. Acute rejection reactions mostly occurred immediately after the interruption of therapy. There was no increased risk associated with a longer break in treatment. Interruption of therapy during the first period of immunosuppressive treatment, that is approximately 3 months after transplantation, was associated with the lowest risk of rejection. As a result of our experience in 25 cases it appears that the temporary suspension of immunosuppressive therapy is associated with little risk and avoids a drug intoxication as well as reduces the mortality due to infection.


[Progress in isolation technic of islets of Langerhans for transplantation].

An improved technique is described which yields up to 6,000 isolated islets of Langerhans from the pancreas of rats. The quotients for insulin/amylase and insulin/protein were estimated in vitro. For in vivo testing isologous transplantations of islets were performed. Several parameters such as total body-weight, blood sugar levels, insulin i.s., glucose-tolerance and glucose-assimilation tests demonstrated that the islets still retained their normal function despite the isolation procedure.


Management of the patient with myocardial infarction.

Coronary atherosclerotic heart disease and myocardial infarction constitute an epidemic in this century, mandating that the primary care physician be familiar with their recognition and management. However, in recent decades, an improved understanding of pathophysiologic alterations, an enormous advance in technology and significant accomplishments in pharmacology and operative procedures have virtually revolutionized the management of patients with myocardial infarction. Although patterns of care described in this monograph may well be obsolete a few years hence, to be superseded by safer, more precise, more efficient and more cost-effective therapeutic modalities, nevertheless basic principles underlying the management of the patient with myocardial infarction remain as appropriate guidelines. In the hospital phase, efforts should be directed toward enhancing survival, saving myocardium and restoring function. The long-term ambulatory care should be designed to maintain functional capacities, to control symptoms, to retard or arrest the atherosclerotic process thus decreasing the likelihood of recurrent myocardial infarction and/or of sudden cardiac death and to restore the patient to a normal or near-normal life style.


The effects of thyrotropin-releasing hormone on the endocrine pancreas.

TRH has been shown to be present in the pancreas. To examine a possible role for TRH in the control of endocrine pancreatic function, we have studied the effects of TRH on the isolated perfused rat pancreas preparation. Arginine caused release of TRH from the preparation. The mean maximum TRH peak was 85 +/- 12 pg/ml and occurred later than the first phase of glucagon release. Glucagon (2000 pg/ml) did not release TRH from the preparation. There was no detectable basal release of TRH. Glucose did not stimulate release of TRH from the pancreas preparation. TRH (10 ng/ml) by itself had no effect on insulin or glucagon release. TRH enhanced arginine-induced glucagon release; mean summated glucagon was 8228 +/- 1138 (SE) pg/ml compared to controls (4530 +/- 447 pg/ml; P less than 0.01). There was a tendency for TRH to enhance second phase glucose-induced insulin release. Pancreatic physiology is in part regulated by locally acting hormones and TRH may be one of these hormones.


Studies on rat pituitary homografts. II. Effects of thyrotropin-releasing hormone on in vitro biosynthesis and release of growth hormone and prolactin.

A study was made of the effect of TRH, administered in vivo by iv infusion or added in vitro to the incubation fluid, on tissue fragments prepared from rat normotopic and ectopic pituitaries. The latter were 30-day-old pituitary grafts transplanted under the kidney capsule in hypophysectomized animals. No detectable effect of TRH was found with normotopic glands. In contrast, the neurohormone produced a large increase in GH and PRL biosynthesis in the grafts, as revealed by the rise in hormone content as well as increased incorporation of L-[3H]leucine into the two hormones. These effects of TRH 1) are dose related; 2) appear after a latent period of at least 15 min; and 3) persist, although attenuated, for some time after removing the neurohormone. In vitro release of GH and PRL by tissue fragments prelabeled with L-[3H]leucine was studied by following the appearance of the radioactive hormones in the incubation fluid. Exposure to TRH produced a prompt, 2-fold or greater increase in hormone release from grafts but not from normotopic gland fragments. The possible mechanisms whereby pituitary somatotrophs and mammotrophs removed from the influence of the central nervous system increase their responsiveness to TRH stimulation are considered.


Estrogen-induced gonadotropin surges in female rhesus monkeys after pituitary stalk section.

In order to investigate the primary site of action of estradiol, whether pituitary or hypothalamic, gonadotropin responses to estrogen were studied in female rhesus monkeys before and immediately after pituitary stalk section. The estrogen challenge, consisting of either an injection of estradio benzoate (400 microgram) or an implant of three silastic capsules containing 17 beta-estradiol, was initiated on days 2--5 of the menstrual cycle. The estrogen was given not later than 8 h after stalk section. Estrogens induced LH surges in all five animals before and after stalk section. FSH increases were observed in four of five intact and three of four stalk-sectioned animals. Mean FSH and LH levels in three stalk-sectioned animals treated with oil alone did not differ significantly from preinjection controls. These experiments suggest that the locus of estrogens on gonadotropin release in the rhesus monkey may well reside within the pituitary gland itself.


A diurnal melatonin rhythm in primate cerebrospinal fluid.

Melatonin was measured in cerebrospinal fluid (CSF) withdrawn continuously from partially restrained rhesys monkeys. There was a daily rhythm in CSF melatonin with peak night values 2- to greater than 15-fold higher than day values. The increase occurred shortly after lights were turned off, and the decrease occurred soon after lights were turned on. There was substantial variation in the magnitude of the rhythm among animals. However, there was little day to day variation in the rhythm of individual animals studied for 3 or 6 consecutive days. Although the concentration of melatonin in CSF was lower than that in plasma, the changes in CSF melatonin concentrations seemed to reflect large daily changes in plasma melatonin concentrations.


Lobulo-alveolar development of mouse mammary glands is regulated by thyroid hormones.

The mammary glands of euthyroid female C3H/HeN mice undergo a series of morphological changes during development. In glands from immature animals, the epithelial component consists of a sparse ductal system with few branches which fills about one fourth of the fat pad. In the adult virgin gland, the epithelial component fills the fat pad with a highly branched ductal system and a few alveoli. In contrast, glands from adult animals maintained in a hypothyroid state by ingestion of thiouracil since weaning retain the primitive ductal appearance while filling the fat pad. The glands from animals made hyperthroid by adding 2 micrograms T4/ml drinking water have extensive lobulo-alveolar development. Glands from animals made hypothyroid during 7 weeks of involution after lactation have the same degree of deveopment as the euthyroid controls. When explants of tissue from adult hypothyroid virgin animals are cultured in serum-free medium containing insulin, hydrocortisone, and PRL, the specific milk protein, alpha-lactalbumin, is induced. The level of alpha-lactalbumin, measured as lactose synthetase activity, found per ng epithelial DNA is the same as that found in explants from glands of euthyroid virgins. These results suggest that thyroid hormones, in concert with PRL, play an important role in the regulation of development of the mouse mammary gland. Decreased levels of thyroid hormones in the serum result in retarded growth of the ductal system and little or no alveolar development. However, the resulting epithelial component of glands from hypothyroid mice is fully capable of differentiating in vitro when exposed to the proper hormonal environment.


Hypoandrogenism and abnormal regulation of gonadotropin secretion in rats fed a low protein diet.

The function of the hypothalamic-pituitary-testicular axis was evaluated in rats fed a low protein diet for 4 weeks beginning at 21 days of age. Compared to control, the low protein group had decreased seminal vesicle and prostate weights as well as decreased testicular testosterone output in vitro, although serum testosterone was not different. The low protein group showed no consistent alterations in serum LH (basal, post-LHRH, and postcastration) compared to control although serum FSH (basal and post-LHRH) was lower in the low protein group. Despite this lower basal FSH, the low protein group had supranormal serum FSH after castration. Seminiferous tubule diameter and testicular histology were normal in the low protein group although testicular androgen-binding protein was absent. Testicular androgen-binding protein was also undetectable in a modestly food-restricted control group which had normal testicular size, testicular histology, androgen output, and serum FSH. This finding suggests that loss of testicular androgen-binding protein may be a sensitive sign of undernutrition. We conclude that rats fed a low protein diet have hypoandrogenism, normal testicular histology, and supranormal FSH after castration despite subnormal basal FSH. The latter combination suggests overproduction of an FSH inhibitor of testicular origin.


Evidence that prolactin stimulates alpha-lactalbumin production in mannary tissues from premenarcheal rhesus monkeys.

PRL was found to stimulate marked increases in alpha-lactalbumin production in six of eight specimens of mammary tissue from premenarcheal rhesus monkeys, and a lesser increase was seen in a seventh. Even without added PRL, low concentrations of this milk protein (mean total alpha-lactalbumin production, 1.9 ng/dish) were released into the organ culture medium bathing these relatively immature tissues; most of the epithelial elements were ductal. Under similar conditions, significantly higher concentrations of alpha-lactalbumin (mean total production, 23.4 ng/dish) were released from tissues of sexually mature animals in which lobulo-alveolar elements were abundantly present in addition to ducts. When tissues from premenarcheal animals were exposed to ovine PRL, alpha-lactalbumin concentration in medium and tissue homogenates were increased significantly. Overall, mean total alpha-lactalbumin production rose to 12.9 (P less than 0.02) and 40.5 (P less than 0.02) ng/dish in response to 100 and 1000 ng/ml ovine PRL, respectively. In those cases in which both medium and tissue homogenates were analyzed, increases were parallel. These findings indicate that PRL has a lactogenic effect on mammary tissue from sexually immature and mature rhesus monkeys.


Enzyme immunoassay of angiotensin I.

A very sensitive and specific enzyme immunoassay has been developed for angiotensin I. Angiotensin I was coupled to beta-D-galactosidase by a novel cross-linking reagent, N-(meta-maleimidobenzoyloxy)succinimide. No decrease in the enzyme activity was observed during the coupling procedure. In the angiotensin I-beta-D-galactosidase conjugate, 0.39 mol immunoreactive angiotensin I/mol enzyme were present. A competitive assay with the enzyme-labeled angiotensin I was performed. Antibody-bound and free labeled antigen were separated from each other by the second antibody method, and the enzyme activity of the former was estimated. Using this assay, angiotensin I could be detected in the range of 1.2--50 pg. The sensitivity was 4.5-fold higher than that of the usual RIA. This assay distinguished clearly angiotensin I from angiotensin II, angiotensin III, and (Sar1, Ile8)-angiotensin II. The present method was applied to measure PRA in dogs; the results correlated fairly well with those obtained by the RIA (r = 0.94).


Effects of neuropeptides on adenohypophyseal hormone response to acute stress in male rats.

The effects of bombesin and other unrelated oligopeptides on hormonal changes induced by stress were studied in conscious adult male rats. Restraint in the cold for 1 h increased plasma corticosterone and PRL levels and decreased GH values but had no effect on LH levels. Bombesin (5 microgram), given intracerebroventricularly (ivt) before stress, inhibited the PRL rise without affecting corticosterone, GH, or LH response. A complete blockade of PRL rise was observed with doses of bombesin ranging from 5 microgram to 100 ng ivt, regardless of the duration (15, 30, 45, or 60 min) or the nature (cold exposure or restraint at room temperature) of the stressor agents. Bombesin was 10(3) more potent as a PRL inhibitor when given ivt than when given iv, and its ivt effect was not reversed by naloxone (1 or 10 mg/kg). Among other unrelated peptides tested (beta-endorphin, neurotensin, substance P, and TRH; 5 microgram ivt), only neurotensin decreased plasma PRL levels in rats subjected to restraint in the cold for 1 h. These results show that in conscious male rats, centrally administered bombesin has a very potent and long acting inhibitory effect on PRL release induced by acute stress. Since a bombesin-like peptide has been found in rat brain, its physiological role in PRL regulation remains to be elucidated.


Between-ovary interaction in the regulation of follicle growth, corpus luteum function, and gonadotropin secretion in the primate ovarian cycle. III. Temporal and spatial dissociation of folliculogenesis and negative feedback regulation of tonic gonadotropin release after luteectomy in rhesus monkeys.

This study was designed to examine effects of previous ovarian status on subsequent follicle growth and the role of between-ovary communication in the regulation of folliculogenesis and gonadotropin secretion during the primate ovarian cycle. Responses to luteectomy were compared in two groups of rhesus monkeys. In the first, follicle growth and corpus luteum function had been constrained chronically to a single ovary by hemiovariectomy performed 66--258 days earlier; the second group was composed of intact monkeys that underwent contralateral wedge resection at luteectomy. In each group, luteal ablation was followed by a prompt fall in serum progesterone levels, a premature onset of menses, and the next preovulatory gonadotropin surges 14.7 +/- 1.1 or 15.4 +/- 1.4 days later (mean +/- SE; P greater than 0.25). Although the patterns of circulating estradiol before and after ablation in each group were superimposable, luteectomy in monkeys lacking a contralateral ovary was followed by a large (2- to 4-fold) and prolonged (7--10 days) increase in serum FSH, whereas in monkeys with two ovaries, serum FSH levels exhibited only a small short-lived rise. The findings indicate that 1) prior chronic constraint of ovarian function to a single ovary did not alter the overall time course of new follicle growth culminating in ovulation after luteectomy; 2) the contralateral ovary provided the principal negative feedback regulation of gonadotropin secretion for some time after luteectomy even though it may not have been the exclusive site of new follicle growth; 3) whereas the ability of the luteectomized ovary to regulate tonic gonadotropin secretion was temporarily impaired, its ability to support the customary temporal pattern of follicle growth after luteal ablation was not; 4) some (contralateral) ovarian factor other than estradiol or progesterone apparently made a major contribution to the regulation of FSH secretion after luteectomy; and 5) folliculogenesis culminating in ovulation from a single follicle and the negative feedback regulation of tonic gonadotropin secretion in some circumstances may occur concurrently but separately on opposite ovaries or may occur at different times within the same ovary.


Comparison between the thyrotropin response to thyrotropin-releasing hormone in summer and that in winter in normal subjects.

A comparison was made between the thyrotropin (TSH) response to 500 microgram thyrotropin-releasing hormone (TRH) in summer and that in winter in ten healthy normal adults living in Supporo. The serum resin triiodothyronine (T3) uptake (RT3U), thyroxine (T4) and T3 levels were also measured. While the TSH response to TRH in summer was similar to that in winter, serum T3 concentration and free T3 index were significantly higher in winter than in summer, associated with the similar values in RT3U and T4 levels in serum. Independently measured 86 specimens (43 in summer and 43 in winter) from normal adults living in the same district also showed a significant increase in serum free T3 index as well as a slight elevation of serum T3 concentration in winter but not in serum T4 level. These results indicate that the primary change in cold winter would be the stimulation of peripheral conversion of T4 to T3 rather than the activation of hypothalamo-pituitary-thyroid axis. The relevance of this interpretation was discussed.


Effect of active and passive immunization with TRH on plasma TSH response to propylthiouracil.

The role of thyrotropin-releasing hormone (TRH) in the secretion of TSH from the anterior pituitary was investigated in rats by active and passive immunization with TRH. The plasma TSH response to propylthiouracil (PTU) in TRH-bovine serum albumin (BSA)-immunized rats was significantly lower than that of BSA-immunized or non-immunized rats. Similarly, the increased plasma TSH level following PTU treatment was significantly suppressed after iv injection of antiserum to TRH. However, the decline in plasma TSH levels was not complete. The results of the present study indicate, at least in part, the physiological significance of endogenous TRH in the regulation of pituitary TSH secretion.


Cerebellar stimulation in alumina-gel monkey model: inverse relationship between clinical seizures and EEG interictal bursts.

The efficacy of cerebellar stimulation was addressed in a chronic monkey model (N = 12) of spontaneous focal motor and secondarily generalized seizures using 24 hr seizure frequency monitoring and all-night EEG recording. The anterior cerebellar vermis was stimulated employing parameters similar to those used in man, 10 Hz, 1 msec pulses, 10 min on, 10 off, at an average current of 2.0 mA. Six weeks pre- and post-base-line periods were compared to a stimulation period of the same length. The results contribute to a clarification of conflicting findings of previous researchers by revealing an inverse relationship between seizure frequency and interictal EEG bursts during the weeks of stimulation. Seizure frequency increased significantly and interictal bursts decreased. Both of these effects (especially the former) were evident in the post-stimulation period, but for different reasons than hypothesized for the period of stimulation. Whereas the therapeutic value of cerebellar stimulation on seizures may be in question, its utilization in the study of mechanisms of epilepsy may be warranted.