Medulloblastoma: treatment results and effect on normal tissues.

Thirty-three children under age 20 with medulloblastoma, treated between 1962 and 1976, at the University of California and the Claire Zellerback Saroni Tumor Institute of Mount Zion Hospital, San Francisco, were retrospectively studied. A relationship between dose and local control rate was suggested by an improved five-year survival in those patients receiving doses greater than 5000 rads to the posterior fossa. The posterior fossa, either alone or with the spinal cord, was the most frequent site of failure. Results of re-irradiation for failure were encouraging and no significant complications were noted. A study of the effects of craniospinal irradiation on the hematopoietic and immune system demonstrated a marked decrease in the peripheral lymphocyte population at the completion of therapy and suggested a functional impairment of the remaining lymphocytes. Other side effects of irradiation included suppression of the hypothalamic-pituitary axis and one instance of brain necrosis. Current treatment policy and proposals for future modifications are discussed.


Radiation in multidisciplinary treatment of children with malignant ovarian tumors.

Thirteen girls with malignant ovarian tumors treated at Memorial Hospital from August 1971 through October 1975 with extensive surgery, multi-drug chemotherapy (T-2 protocol) and postoperative radiation were reviewed. The plan of radiation was to deliver 3,000 rads to the whole abdomen and boost the para-aortic and pelvic area with 1,500 rads in five to six weeks. In only two patients was the radiation conducted as planned. In others the treatment was either interrupted or discontinued due to complications. Four of the thirteen patients are dead. The cause of death in two of these patients was distant metastases; the two other patients died without evidence of tumor due to treatment complications. Three of the nine patients who are alive without evidence of disease have late complications. Following radical surgery, lower doses of radiation given concomitantly with T-2 protocol chemotherapy seem to be sufficient for controlling the disease.


Irradiation alone or in combination with surgery in stage IB and IIA carcinoma of the uterine cervix: A nonrandomized comparison.

This is a report of a nonrandomized comparison of treatment results of 244 patients with stage IB carcinoma of the uterine cervix treated by radiation alone and 92 treated with preoperative radiation and surgery and 77 patients with stage IIA treated by radiation alone and 24 treated with a combination of radiation and surgery. The techniques of irradiation and types of operation are described in detail. The five-year tumor free actuarial survival for the patients with stage IB treated either with irradiation alone or combined with surgery was approximately 85% and the ten-year survival, 78%. For stage IIA the tumor free actuarial five-year survival without tumor was 73% and for ten years, 60%. In the 244 patients treated with radiation alone, there were ten central failures (4%) usually combined with distant metastasis. Further, 16 of these patients (6.5%) developed parametrial recurrence, in all but one instance associated with distant metastasis. In the 92 patients with stage IB treated with combined therapy, there were three local recurrences (3.8%), two of them combined with parametrial failures and six parametrial recurrences (6.5%), all of them concomitant with distant metastasis. Of the 77 patients with stage IIA treated by irradiation alone, there  was one central recurrence alone and five local and parametrial recurrences, all of them associated with periaortic nodes or distant metastasis. Four additional patients had parametrial recurrences only concurrent with distant metastasis. Of the 24 patients treated with irradiation and surgery, there were two parametrial recurrences combined with distant metastasis (8.2%). There was no significant difference in the survival or recurrence rate of the patients treated with either method. In the group treated with combined therapy, patients with stage IB who showed evidence of microscopic residual tumor after irradiation had a failure rate of approximately 42% (8/18) in contrast to only 8.6% (6/70) in those with negative specimens. In stage IIA there were three failures in eight patients with residual tumor in the specimen in contrast to only two of 16 with negative specimens (12.5%). Major complications were comparable in both groups (radiation alone approximately 8.7% and irradiation combined with surgery approximately 14%), the difference is not statistically significant. The most frequent minor complication in patients treated with radiation alone was vaginal fibrosis (30 patients--9%) or vaginal vault necrosis (10 patients--3%).


Evaluation of nutritional indices as prognostic indicators in the cancer patient.

The nutritional and immunological status of 140 malnourished cancer patients who were to receive intravenous hyperalimentation (IVH) were evaluated prior to treatment with IVH and chemotherapy (CMX), surgery or radiation therapy (XRT). Subsequently, these indices were correlated with responses to treatment. Cell-mediated immunocompetent (CMI+) patients (41%) had lost an average weight of 10 +/- 2.2 lbs., while cell-mediated immunoincompetent (CMI-) patients (59%) had lost an average weight of 31 +/- 2.5 lbs. (p less that 0.05). Total lymphocyte count (TLC) averaged 1290 +/- 250 cells/mm3 in CMI+ patients and 900 +/- 140 cells/mm" in CMI- patients (p less than 0.05). Serum albumin concentration (SA) was 3.6 +/- 0.06 g/dl in CMI+ patients and 3.3 +/- 0.05 g/dl in CMI- patients (p less than 0.05). In the CMI+ group, 49% had a positive response to CMX, whereas only 27% of the CMI-group responded to CMX (p less than 0.01). In the surgery group, 88% of the CMI+ patients, but only 65% of CMI- patients, had uncomplicated postoperative recovery periods. Although nutritional repletion with IVH was undertaken in all patients, those patients with initial CMI- responses had greater body weight loss, lower SA and TLC and either a decreased response to CMX or an increase in postoperative morbidity and mortality when compared with patients who initially had CMI+ responses.


Results of treating stage III carcinoma of the breast by primary radiation therapy.

One hundred sixteen patients with stage III carcinoma of the breast were treated by primary radiation therapy. The 5-year actuarial survival and relapse-free survival were 25% and 22%, respectively. The 5-year actuarial probability of local tumor control for the entire group was 64%. In patients undergoing an excisional biopsy and an interstitial implant of the primary tumor area, local control was 100%. In patients who had either an excisional biopsy or an implant, the 5-year actuarial probability of local control was 77% and 76%, respectively. In contrast, in patients having neither an excisional biopsy nor an implant, local control was only 41%. In patients receiving a total dose of greater than 6000 rad, from external beam treatment or from external beam plus an interstitial implant, the local control was 78% compared to 39% in patients receiving a total dose of less than 6000 rad. Forty-one patients received some form of adjuvant therapy. Both local control and relapse-free survival were improved in patients receiving chemotherapy as the sole adjuvant and in patients receiving chemotherapy combined with an endocrine ablative procedure. However, patients treated with only an endocrine ablative procedure had no improvement in survival nor in local control. These results indicate that primary radiation therapy can provide local control in a high proportion of patients with stage III carcinoma of the breast and suggest that chemotherapy is effective in improving both local control and survival in these patients.


Results of total body irradiation in the treatment of advanced non-Hodgkin's lymphomas.

Total body irradiation (TBI) was used as primary therapy for 58 previously untreated patients with Stage III or IV non-Hodgkin's lymphoma (NHL). 150 rad was administered, with 15 rad fractions twice a week, with careful monitoring of hematologic status. Thrombocytopenia was the most frequent complication, which resolved in all except 4 patients. Survival at 8 years was 52%, with 14% relapse-free survival. Patients with nodular histology had a more favorable prognosis than those with diffuse histology (median relapse-free survival of 24 vs. 12 months). There were 2 cases of erythroleukemia, which occurred after combination chemotherapy was given for relapse. Though TBI can offer complete remission and extended survival in advanced NHL, most patients eventually relapse and it should not be considered as a curative mode of therapy.


In vivo and in vitro studies of experimental ovarian adenocarcinoma in rats.

When 7,12-dimethylbenz[a]anthracene-impregnated sutures were directly applied to the ovarian parenchyma of 8-week-old Sprague-Dawley rats (the clipping method), adenocarcinomas developed in 29 (39%) of the 75 rats during the 50-week observation period. When 20-methylcholanthrene was used, adenocarcinomas developed only in 1 (3%) of the 31 rats. Thus, the clipping method using 7,12-dimethylbenz[a]anthracene is satisfactory as an animal model of ovarian adenocarcinoma which comprises 85 to 90% of human malignant ovarian tumors. On the other hand, attempts were made to isolate cloned cell lines from these experimental ovarian adenocarcinomas in vitro, and two cloned cell lines were obtained. They were epithelioid and produced undifferentiated adenocarcinomas by back-transplantation into isologous newborn rats.


DNA-protein cross-linking by chemical carcinogens in mammalian cells.

The induction of DNA cross-linking in mammalian cells by various carcinogens was investigated by the method of alkaline elution. A dose-dependent increase in DNA cross-linking was seen following exposure of human fibroblasts to N-acetyoxy-2-acetylaminofluorene and following exposure of mouse embryo cells to 7,12-dimethylbenz[a]-anthracene. No cross-link effect was seen following treatment with N-methyl-N'-nitro-N-nitrosoguanidine, benz-[a]anthracene, benz[A]anthracene-5,6-dihydroepoxide, or metabolic inhibitors. The cross-linking appeared to be DNA-protein in nature since proteinase treatment removed the effect. DNA single-strand breaks were also induced by several of these agents in the case of N-acetoxy-2-acetylaminofluorene and N-methyl-N'-nitro-N-nitrosoguanidine, approximately 70 to 90% of these breaks were rejoined after an 18-hr incubation in fresh medium, whereas repair of the cross-links induced by N-acetoxy-2-acetylaminofluorene was slight at this time.


The cytotoxicity of gamma-L-glutaminyl-4-hydroxybenzene for cells that contain tyrosinase, a study of melanocytes in the hair follicle of the mouse.

The s.c. administration of gamma-L-glutaminyl-4-hydroxybenzene (GHB) to neonatal black mice produced a prompt, generalized, and selective swelling and lysis of the melanocytes of the hair follicles. The findings indicate that this cytotoxic effect was dependent upon the intracellular activation of GHB by tyrosinase. Supportive of this conclusion were: (a) an absence of comparable cytological alterations in adjacent keratinocytes; (b) a lack of response by melanocytes of albino mice; and (c) patterns of deficient pigmentation produced by GHB in juvenile black mice, suggesting that susceptible follicles were those in the tyrosinase-producing growth phase. The administration of GHB also induced condensation, or "apoptosis," of individual follicular keratinocytes of both black and albino mice and in the melanocytes in the latter. This response was apparently independent of tyrosinase. It was transitory and without appreciable effect on hair growth. The findings further characterize the selective cytolytic properties of GHB for mammalian cells that possess tyrosinase and suggest a potential for this natural compound as a chemotherapeutic agent against melanocarcinoma.


Use of molecular hybridization to detect type D retrovirus markers in rhesus placentas and other tissues.

We have shown previously that approximately 20% of the Mason-Pfizer virus (MPV) genome is present as endogenous provirus in rhesus monkeys. We report here that several full-term rhesus placentas examined contain additional MPV proviral sequences in their DNA. Competitive molecular hybridization experiments demonstrated that some of these placentas also contain RNA complementary to the entire MPV 60 to 70S RNA genome. Examination of internal organs of rhesus monkeys captured in the wild also revealed the presence of additional MPV proviral sequences and expression of MPV RNA in some tissues. These results provide further evidence that MPV is being transmitted via a non-germ line mechanism in the rhesus population and now demonstrate the placenta as a good source for the identification of retrovirus transcriptional products and proviral DNA.


Comparative randomized study of protected environment plus oral antibiotics versus oral antibiotics alone in neutropenic patients.

A series of 24 patients with severe neutropenia, most of whom had acute myeloblastic leukemia, were treated in an isolation unit with oral nonabsorbable antibiotics and were compared to 21 similar patients receiving oral antibiotics alone. The frequency of bacterial infections was lower in the patients receiving both isolation and oral antibiotics compared to the patients who received only oral antibiotics. The responses to chemotherapy in terms of remission rates were identical for the two groups.


cis-Dichlorodiammineplatinum(II) and hexamethylmelamine in the treatment of non-oat cell lung cancer: a pilot study of the Southeastern Cancer Study Group.

Fifty-one patients with metastatic non-oat cell carcinoma of the lung were treated with a combination of cis-dichlorodiammineplatinum(II) and hexamethylmelamine. The overall response rate (all cell types) was 16%. Four of 20 patients with adenocarcinoma had a partial response and an additional five patients classified as having stable disease had tumor regression less than 50%. The median survival of responders and of those with stable disease (all types) was 8 and 7 months respectively, significantly longer than the median survival of patients who progressed (median survival, 2 months [P less than 0.05]). The major dose-limiting toxicity was nausea and vomiting in over half of the patients; hematologic toxicity and peripheral neuropathy were the other adverse effects of the combination.


Controlled clinical studies with an antidote against the urotoxicity of oxazaphosphorines: preliminary results.

A randomized study in 20 patients with cancer was carried out to test the clinical efficacy of sodium-2-mercaptoethane sulfonate (ASTA D-7093; mesnum) as an agent to prevent urotoxic side effects (in particular, hemorrhagic cystitis) during cytostatic therapy with the oxazaphosphorines cyclophosphamide and ifosfamide. Eleven patients received mesnum iv and nine patients received a standard prophylaxis. The frequency of microhematuria was significantly lower in the patients receiving mesnum. A slight microhematuria was observed in one patient. With the standard prophylaxis, all nine patients receiving single-agent therapy with ifosfamide or cyclophosphamide had hematuria and three of these had macrohematuria. According to the available results, a daily mesnum dose of 60% (wt/wt) of the ifosfamide or cyclophosphamide dose is recommended. This dose should be divided into three equal fractions. The first administration should be given concurrently with the cytostatic agent and the subsequent two administrations at 4 and 8 hours after administration of the cytostatic agent. Significantly higher doses of mesnum (eg, 133% of the cyclophosphamide or ifosfamide doses) lead to gastrointestinal disorders, which are easily reversible.


Production, purification, and properties of dextransucrase from Leuconostoc mesenteroides NRRL B-512F.

The production of dextransucrase from Leuconostoc mesenteroides NRRL B-512F was stimulated 2-fold by the addition of 0.005% of calcium chloride to the medium; levansucrase levels were unaffected. Dextransucrase was purified by concentration and dialysis of the culture supernatant with a Bio-Fiber 80 miniplant, and by treatment with dextranase followed by chromatography on Bio-Gel A-Fm. A 240-fold purification, with a specific activity of 53 U/mg, was obtained. Contaminating enzyme activities of levansucrase, invertase, dextranase, glucosidase, and sucrose phosphorylase were decreased to non-detectable levels. Poly(acrylamide)-gel electrophoresis of the purified enzyme showed only two protein bands, both of which had dextransucrase activity. These bands also gave a carbohydrate stain, indicating that the dextransucrase could be a glycoprotein. Acid hydrolysis, followed by paper chromatography, of the purified enzyme showed that the major carbohydrate was mannose. Concanavalin A completely removed dextransucrase activity from solution, confirming the mannoglycoprotein character of the enzyme. Dextransucrase activity was not altered by the addition of 0.008-4 mg/ml of dextran, but its storage stability was increased by the addition of 4 mg/ml of dextran. As previously shown by others, the activity of dextransucrase was decreased by EDTA, and was restored by the addition of calcium ions. Zinc, cadmium, lead, mercury, and copper ions were inhibitory to various degrees.


alpha-L-iduronidase, beta-D-glucuronidase, and 2-sulfo-L-iduronate 2-sulfatase: preparation and characterization of radioactive substrates from heparin.

Radioactive disaccharide substrates for alpha-L-iduronidase, beta-D-glucuronidase, and 2-sulfo-L-iduronate 2-sulfatase have been prepared from heparin by deaminative cleavage followed by reduction with NaBT4. Six disaccharides were isolated from this reaction mixture and identified. Acid hydrolysis of the major disaccharide, O-(alpha-L-idopyranosyluronic acid 2-sulfate)-(1 linked to 4)-(2,5-anhydro-D-mannitol-l-t 6-sulfate (IdAs--Ms), produced 48% of O-(alpha-L-idopyranosyluronic acid)-(1 linked to 4)-(2,5-anhydro-D-mannitol-l-t 6-sulfate) (IdA--Ms) and 25% of O-(alpha-L-idopyranosyluronic acid)-(1 linked to 4)-2,5-anhydro-D-mannitol-l-t. The most-sensitive substrate for determining alpha-L-iduronidase activity was IdA--Ms which, when incubated with leucocyte and skin-fibroblast homogenates prepared from patients having a deficiency of alpha-L-iduronidase (Mucopolysaccharidosis Type I; MPS-I), was hydrolysed to yield 2,5-anhydro-D-mannitol-l-t 6-sulfate at a rate 50-times less than that found for normal control-preparations. Similarly, O-(beta-D-glucopyranosyluronic acid)-(1 linked to 4)-(2,5-anhydro-D-mannitol-l-t 6-sulfate) was degraded by whole-cell homogenates prepared from beta-D-glucuronidase-deficient (Mucopolysaccharidosis, Type VII) fibroblasts, to yield 2,5-anhydro-D-mannitol-l-t 5-sulfate at a rate 60-times less that that found for MPS-I and normal control-preparations. IdAs--Ms was degraded by 2-sulfo-L-iduronate 2-sulfatase at a rate more than 45-times greater than that found for O-(alpha-L-idopyranosyluronic acid 2-sulfate)-(1 linked to 4)-2,5-anhydro-D-mannitol-l-t. C-6 Sulfation of the anhydro-D-mannitol-l-t residue is an important structural determinant in the mechanism of action of both alpha-L-iduronidase and 2-sulfo-L-iduronate 2-sulfatase on disaccharide substrates.


Studies on metabolic activation of vinyl chloride in Drosophila melanogaster after pretreatment with phenobarbital and polychlorinated biphenyls.

It is known that vinyl chloride is metabolized by the mixed function oxygenase system in the liver to reactive mutagenic and carcinogenic metabolites. This metabolic activation was studied in Drosophila melanogaster by measuring the uptake of 14C from labelled vinyl chloride in different strains and with different pretreatments with phenobarbital and polychlorinated biphenyl (PCB) Clophen A50), well known inducers of cytochrome P-450. In accordance with previously obtained data on vinyl chloride induced sex linked recessive lethals, it was shown that pretreatment with inducers increased the uptake of labelled compound up to ten times. There was, however, a marked difference in response between the five strains used. In particular, the strain Hikone, known to be resistant to insecticides, had a comparatively high initial radioactivity from vinyl chloride without any pretreatment, but it was not or insignificantly inducible with phenobarbital or PCB. Crosses between Hikone and an inducible strain indicated essentially a dominance for the Hikone genotype. Tests on inducible strains showed the same response to phenobarbital by 2 h old larvae and adult male and females. Dimethylsulphoxide (DMSO) used as a solvent decreased both the initial uptake of 14C and particularly the induction by PCB. The use of Tween 80 as an emulsifier did not have such an effect. It is emphasized that the interstrain variation in metabolic activation and inducability has to be taken into consideration in order to optimize the use of Drosophila for mutagenicity testing. This variation also opens up new possibilities of analyzing the mixed function oxygenase system biochemically and genetically.


Sensitivity differences displayed by Drosophila melanogaster larvae of different ages to the toxic effects of growth on media containing aflatoxin B1.

Using Drosophila melanogaster, the relative sensitivities of various larval stages to the toxic effects of growth on media supplemented with either 0.44 or 0.88 ppm aflatoxin B1 (AFB1) were determined. Two strains of fruit flies were tested: strain A-11 which is relatively resistant to AFB1 induced toxicity, and strain A-9 which is quite sensitive. Eggs, mid-first, mid-second and early-, mid- and late-third instar larvae were transferred onto AFB1 media and allowed to complete larval and pupal development and eclose as adults. At the 0.44 ppm concentration, strain A-11 showed no effect, while only first instar larvae of strain A-9 showed significant mortality rates for first instar larvae, but the A-9 larvae die at higher rates than the A-11 larvae. In addition, second and third instar larvae of strain A-9 show significant mortality rates when grown at 0.88 ppm AFB1, while these stages are not affected in strain A-11.


[Role of the Raphe Magnus nucleus in morphine analgesia : studies with intracerebral microinjections in the rat].

Microinjections of low concentration of morphine (5 micrograms) into the nucleus Raphé Magnus of the Rat produce a strong analgesia that can be reversed by systemic naloxone, an opiate antagonist. The administration of naloxone (5 micrograms) into the Raphé Magnus considerably reduces the effects of intravenous morphine. The effects of microinjections of morphine are strongly reduced by Cinanserin, suggesting a role for serotoninergic mechanisms in morphine analgesia.


[I and i antigens of human erythrocytes : their detection by means of immunofluorescence technic and their changes during ontogenesis].

I and i antigens were detected by indirect immunofluorescence on human erythrocytes. 80% of the cord blood cells were brightly fluorescent with anti-i, 15% only with anti-I. In adult blood, this ratio is reversed, 2% only of the cells being intensively labelled with anti-i. The percentages characteristic of adult blood are established within the first 8 months of life.


[Criteria for the evaluation of the antirheumatic (antiinflammatory) drugs in man].

Comparison of 5 antiinflammatory drugs and placebo in a double blind, short term, cross-over trial conducted in patients suffering from rheumatoid arthritis. 15 measurements were used and studied. With classical unidimensional statistical tests it was not possible to discriminate active drugs between them. Multidimensional analysis (correspondance analysis and discriminant analysis) are more sensitive and allow us to distinguish active drugs with only 11 criterion.


[Increased contents of phenylethanolamine, m-octopamine and p-octopamine in the hypothalamus and brain stem of spontanously hypertensive rats (S.H.R. Kyoto)].

Phenylethanolamine, p-octopamine and m-octopamine contents were determined in the hypothalamus and the brain stem of spontaneously hypertensive Rats (S.H.R. Kyto) and the corresponding controls (Wistar Kyoto). In three-week-old Rats, phenylethanolamine and p-octopamine are found to be present in S.H.R. hypothalamus and brain stem at concentrations twice as high as compared to Wistar Kyoto Rats. The amount of m-octopamine is 5-fold higher in the brain stem of S.H.R. Rats as compared to Wistar Kyoto Rats.


[Evidence of a circadian sequence of photogonadsensitivity in the ferret].

Using "skeleton" photoperiods with a primary photofraction of 3 hrs. 30 min. and secondary photofraction of 30 min. located at various intervals after the onset of darkness, the occurrence of a photoinducible phase in the androgenic function has been evidenced in the Ferret. This photo-inducible phase seems to be located around 12 hrs. after the onset of the primary photofraction, since the ferrets who received the 30 min. photofraction 8 hrs. after the onset of darkness responded with a maximal increase in plasma testosterone.


Baroreceptor control of atrioventricular conduction in man.

Although human baroreflexes are known to exert a powerful physiological control on heart rate, little information exists on the physiological control they exert on the atrioventricular conduction system. In 11 normotensive subjects with normal atrioventricular conduction, we altered baroreceptor activity by injection of pressor and depressor drugs (phenylephrine and trinitroglycerin) and recorded mean arterial pressure (MAP, catheter measurements), R-R interval, and pre-His and post-His intervals (A-H and H-V, His bundle recording). With the subjects in sinus rhythm, increasing MAP by 21+/- 1 mm Hg caused a marked lengthening (250 +/- 28 msec), and decreasing MAP by 17 +/- 2 mm Hg a marked shortening (142 +/- 16 msec) of the R-R interval. There was little change in the A-H interval and no change at all in the H-V interval. However, when the R-R interval was kept constant in these subjects by atrial pacing, a similar increase and decrease in MAP caused, respectively, a marked lengthening (49 +/- 6 msec) and shortening (19 +/- 3 msec) of the A-H interval, although the H-V interval remained unaffected. Thus physiological ranges of baroreceptor activation have a marked influence on the atrioventricular node but apparently not on the ventricular portion of the atrioventricular conduction system. This influence is unmasked when pacing prevents the baroreceptor influence on the sinoatrial node.


Effects of nitroglycerin on echocardiographic measurements of left ventricular wall thickness and regional myocardial performance during acute coronary ischemia.

The effects of nitroglycerin on regional left ventricular performance, assessed by echocardiographic techniques, were investigated in anesthetized, open-chest dogs during acute myocardial ischemia. During transient occlusion of the left anterior descending coronary artery, there was end-diastolic thinning and marked reduction in systolic thickening in the central ischemic zone. Similar changes of lesser degree were noted in the border zone. The normal zone was unaffected. Infusion of nitroglycerin during ischemia in dosages of 2.5--50 microgram/kg/min reduced left ventricular end-diastolic pressure without changing the abnormalities of systolic wall thickening. Effects of bolus injections of 20 and 50 microgram/kg of nitroglycerin were similar, although this also lowered aortic pressure. In a subgroup of animals in which nitroglycerin infusion was unaccompanied by tachycardia, there was also no evidence that ischemic dysfunction was altered. We conclude that nitroglycerin does not improve regional myocardial performance in acutely ischemic canine myocardium. The decrease in preload is probably entirely due to the peripheral effects of the agent.


The relationship between endogenous hyperprolactinaemia and plasma aldosterone.

It has been suggested that prolactin is a regulator of aldosterone secretion. In order to test this hypothesis, we measured prolactin, thyrotrophin and aldosterone by radioimmunoassay and plasma renin activity by the radioimmunoassay of angiotensin I in eight normal women before and after the intravenous injection of 200 microgram of thyrotrophin releasing hormone (TRH). Prolactin increased from 4.1 +/- 1.1 ng/ml (mean +/- SE) to a peak of 27.4 +/- 3.8 (P less than 0.005) at 15 min following TRH. Plasma renin activity was not different from control levels (1.0 +/- 0.2 ng/ml/h) during the first hour following the administration of TRH, nor did the plasma aldosterone concentration differ significantly from the control levels (39 +/- 7 pg/ml) during this period. However, with upright posture, an increase in aldosterone (from 31 +/- 3 pg/ml at 1 h to 68 +/- 9 at 2 h, P less than 0.005) and in plasma renin activity (from 0.9 +/- 0.2 ng/ml/h at 1 h to 2.0 +/- 0.5 at 2 h, P less than 0.05) was noted, demonstrating a normal capacity to secrete aldosterone in these subjects. Similarly, no change in aldosterone was seen in nine patients with primary hypothyroidism given TRH, despite the fact that the increase in prolactin was greater than normal. Chronic hyperprolactinaemia was not associated with hyperaldosteronism in six patients with pituitary tumour. These data demonstrate that acutely or chronically elevated serum prolactin levels do not result in increased plasma aldosterone levels in humans.


The effect of lisuride hydrogen maleate in the hyperprolactinaemia-amenorrhoea syndrome: clinical and hormonal responses.

Eleven women with secondary amenorrhoea and hyperprolactinaemia were treated with lisuride, a new semisynthetic ergot derivative. Acute administration of lisuride (100 microgram orally) significantly suppressed serum prolactin (PRL) levels in nine out of eleven subjects. In these nine patients, prolonged treatment with lisuride (50--200 microgram daily) lowered PRL levels into the normal range, menstrual cycles were resumed and eight patients ovulated. Two patients, in whom lisuride failed to lower PRL levels into the normal range, did not resume normal menstruation. These data demonstrate that lisuride may be used clinically to lower PRL levels and to restore cyclic gonadotrophin secretion and ovulation in amenorrhoeic patients with hyperprolactinaemia.


Bacterial and strain specificities in opsonization, phagocytosis and killing of Streptococcus mutans.

Opsonization of Streptococcus mutans, followed by phagocytosis and killing by polymorphonuclear leucocytes has been postulated as an effector mechanism in protection against dental caries. Opsonization was studied by using sera from monkeys immunized with killed Strep. mutans (sero-type c) and compared with sera from sham-immunized monkeys. Antibodies to Strep. mutans (sero-type c) induced maximal phagocytosis and killing of serotypes c and e, and this was significantly greater than with serotypes a and d; there was no significant phagocytosis or killing of serotype b. There was little or no opsonization with Actinomyces viscosus, Lactobacillus casei, Strep, sanguis and Strep. salivarius. The exception was Strep. CHT which showed significant phagocytosis and killing. The results suggest that immunization with the serotype c strain of Strep. mutans might offer protection against four of the five common serotypes of this organism.


Primary meningococcal conjunctivitis in an infant.

A case of primary meningococcal conjunctivitis in an infant was presented and its treatment is discussed. Because of the potential for systemic spread of the infection in the patient, local and systemic therapy was instituted. The current literature on meningococcal conjunctivitis with its potential complications is briefly reviewed.


Proportional polyploidization of 5S RNA genes in the ovary of Drosophila melanogaster mutants containing three 5S RNA gene loci.

The 5S RNA gene content of polyploid cells of the ovary of Drosophila melanogaster has been compared in animals with two or three gene clusters. The amount of 5S RNA genes is exactly proportional to the number of gene clusters as determined by DNA-RNA filter hybridization. In contrast, the number of rDNA genes in endomitotic cells remains constant regardless of different numbers of nucleolus organizer regions (Spear, 1974).


Control of human postprandial pancreatic exocrine secretion: a function of the gastroduodenal region.

Regulation of the exocrine pancreatic secretion elicited by a meal in man is incompletely understood. In this study, we attempted to localize in the gastrointestinal tract areas that control postprandial trypsin secretion and to determine the effects of individual components of jejunal chyme on the meal-stimulated trypsin secretion. Trypsin outputs elicited by ingesting a mixed-nutrient meal and diverting it at the ligament of Treitz proximal to an occlusive balloon were compared with those occurring when the same meal was ingested, diverted at the ligament of Treitz, and immediately reinfused distal to the balloon, a procedure that exposed the entire gastrointestinal tract to chyme. Two different meals, one of semielemental and one of complex nutrients, were used with similar results. Trypsin outputs were similar whether or not jejunal chyme was diverted. In addition, no component--exogenous nutrients or endogenous secretions--of chyme reaching the jejunum after a meal further modified the trypsin secretion elicited by the gastroduodenal segment. This finding suggests that the gastroduodenal segment is sufficient to elicit the entire postprandial trypsin output and is the physiologic determinant of meal-stimulated trypsin secretion.


[Continuous measuring of intracranial pressure (ICP) via epidural pressure transducer (author's transl)].

Continuous epidural monitoring of ICP was performed on 20 patients during a 6 day's period with the model 77450 of Philips. A significant decrease of intracerebral pressure after application of sorbitol and dexamethasone could be demonstrated. Comparing the very low rate of complications with the resulting advantages, the described method can be recommended for controlled treatment of elevated ICP.


[Long-lasting EEG abnormalities in alkylphosphat intoxication (author's transl)].

In the reported case of a severe alkylphosphate poisoning the first EEG-observation at the twentieth day after intake showed marked abnormalities although there was a complete physical and humoral recovery. Progressive normalization of the EEG occured in the following six weeks. Psychopathological symptoms of a mild apathy and slowening of reactions disappeared gradually.


[The 4 c/sec rhythm (case report and differential diagnosis) (author's transl)].

Report on a female patient with a 3 c/sec rhythm. The 3 c/sec rhythm is a special case of the 4 +/- 1 c/sec rhythm. The characteristics of this variant are reviewed in tabulated form. The differential diagnosis has to take into consideration other slow posterior rhythms like the so-called delta-parenrhythmia and delta-parenrhythmia, abortive forms of spike and wave complexes and the abnormal delta-rhythms and sinusoidal delta-rhythms in the infantile EEG.


Serum estradiol, testosterone and dihydrotestosterone in male monkeys treated with testosterone propionate.

Serum estradiol (E2), testosterone (T) and dihydrotestosterone (DHT) were measured in juvenile (pre-pubertal) male rhesus monkeys injected with either 8 mg or 80 mg of testosterone propionate (TP). After one week, the three steroids were elevated and remained essentially unchanged for the duration of the study. There was little difference in serum E2 or DHT when comparing the two groups of steroid-treated monkeys. In contrast, T levels were consistently greater in the animals given the high dosage of TP.


An unusual sex steroid-binding protein in mature male rat liver cytosol.

Mature male rat liver cytosol contains a moderate affinity and capacity estrogen-binding protein in at least a 200-fold higher level than mature female or immature male rat liver cytosol. Binding of estradiol to this protein is very rapid, is stabilized by EDTA, and is inhibited by divalent cations. This is the major binding protein for [3H]estradiol ([3H]E2) in mature male rat liver cytosol, and it has properties clearly distinguishing it from putative liver or uterine estrogen receptors. In addition to binding [3H]E2, this protein seems to rapidly bind a [3H]5alpha-dihydrotestosterone ([3H]DHT) metabolite at the same binding site. The binding of this androgen metabolite is stabilized by EDTA and is inhibited by divalent cations. The binding properties of the [3H]DHT metabolite suggest that these binding sites are not classical androgen receptors. Cytosol binding levels of both the [3H]E2 and the [3H]DHT metabolites change in a similar direction in resonse to endocrine manipulation. The putative liver estrogen receptor level, determined after partial purification (in a redissolved 30% ammonium sulfate-precipitated fraction), seems to change in an opposite direction in response to these same endocrine manipulations.


"Short" loop feedback regulation of hypothalamic and brain thyrotropin-releasing hormone content in the rat and dwarf mouse.

In order to determine whether T4, TSH, or both affect hypothalamic TRH content, primary or secondary hypothyroidism was induced in the rat by thyroidectomy (Tx) or hypophysectomy (Hx), respectively. Two weeks later, rats were treated with T4, TSH, or both for 14--16 days. Tx or Hx significantly decreased hypothalamic TRH content, and T4 treatment restored hypothalamic TRH to normal in the Tx but not in the Hx rats. When TSH was administered simultaneously with T4 to Hx rats, hypothalamic TRH content was restored to normal. Whole brain TRH content was not affected by Tx, Hx, or by the various treatment regimens. Similar experiments were performed in Snell dwarf (dw/dw) mice. Hypothalamic TRH content was significantly decreased when compared to normal litter mates (dw/+). As in Hx rats, T4 administration to dw/dw mice for 16 days did not restore hypothalamic TRH content to normal. Brain TRH content was similar in both groups and was not altered by T4 treatment. It is concluded that T4 affects hypothalamic TRH content, but that TSH is required for this T4 effect. Whole brain TRH, on the other hand, is unaffected by any of these alterations in thyroid-pituitary function.


The developmental pattern of thyrotropin-releasing hormone-degrading activity in the plasma of rats.

The developmental pattern of TRH-degrading activity in rat plasma was determined by measuring the ability of plasma from rats of various ages to degrade TRH into degradation products. From a rate of 0.519 pmol TRH degraded/microliter-1.h-1 on day 8, plasma TRH-degrading activity increased to 18.1 pmol TRH by day 90 in female rats. The increase in rate of formation of proline, a major plasma degradation product, was in very good agreement with the increase in rate of TRH degradation. The major increase in TRH-degrading activity occurred between the third and seventh week of life. A similar pattern of development was observed in male rats; the only significant difference was that plasma from day 90 male rats was approximately 30% more active than that from female rats (P less than 0.002). Daily T3 administration to rats from days 8--26 resulted in a 2-fold increase in plasma TRH-degrading activity (P less than 0.05). The contribution of plasma degradation to the physiological control of TRH activity is not clear, but the magnitude of the increase in plasma TRH-degrading activity (approximately 30-fold) during the maturation of the rat is suggestive of a mechanism of biological significance.


Studies on the mechanism of growth hormone and thyrotropin responses to somatostatin antiserum in anesthetized rats.

An iv administration of 1 ml sheep antiserum to somatostatin (anti-SS) resulted in marked increases of both serum GH and TSH, with a peak 10--20 min after administration in male rats anesthetized with urethane or pentobarbital. Administration of anti-SS had no effect on serum PRL. Ablation of the basal medial hypothalamus abolished the rises of both serum GH and TSH after anti-SS administration. Intravenous injection of 1 ml rabbit antiserum to TRH (anti-TRH) decreased serum TSH levels 15 min after injection, whereas injection of normal rabbit serum did not affect TSH levels. Serum TSH levels did not rise after injection of anti-SS in rats pretreated with anti-TRH. On the other hand, pretreatment with anti-TRH did not affect the basal serum GH levels nor the anti-SS-induced GH release. The enhanced secretion of GH and TSH after anti-SS injections was not blocked by pretreatment with indomethacin, an inhibitor of prostaglandin synthesis. The following conclusions were made: 1) both GH and TSH responses to anti-SS require an intact basal medial hypothalamus; (2) TSH response to anti-SS is mediated by hypothalamic TRH; and 3) the GH response may be mediated by hypothalamic GH-releasing hormone which is not TRH or prostaglandins.


Kinetic and magnetic properties of cobalt(III) ion in the active site of carbonic anhydrase.

Cobalt(III)bovine carbonic anhydrase B was prepared by the oxidation of the cobalt(II) enzyme with hydrogen peroxide and was purified by affinity chromatography. The oxidation reaction is inhibited by specific inhibitors of carbonic anhydrase. The inhibition is explained by the fact that the Co(II)-enzyme . inhibitor complex cannot be directly oxidized by hydrogen peroxide, but has to dissociate to give free Co(II) enzyme which is then oxidized. The Co(III) ion in Co(III) carbonic anhydrase cannot be directly substituted by zinc ions. It can be reduced by either dithionite or BH-4 ions to give, first, their complexes with the Co(II) enzyme, and upon their removal, a fully active Co(II) enzyme. Cyanide and azide bind to cobalt(III) carbonic anhydrase with similar rate constants of 0.060 +/- 0.005 and 0.070 +/- 0.007 M-1 S-1 respectively. These rates are faster than those found for Co(III) inorganic complexes. The Co(III) ion in both Co(III) carbonic anhydrase and Co(III) carboxypeptidase A was found to be diamagnetic, indicating a near octahedral symmetry.


The primary structure of the coat protein of the broad-host-range RNA bacteriophage PRR1.

The complete amino acid sequence of the coat protein of RNA bacteriophage PRR1 is presented. After thermolysin digestion, 26 peptides were isolated, covering the complete coat protein chain. Their alignment was established in part using automated Edman degradation on the intact protein, in part with overlapping peptides obtained by enzymic hydrolysis with trypsin, pepsin, subtilisin and Staphylococcus aureus protease, and by chemical cleavage with cyanogen bromide and N-bromosuccinimide. To obtain the final overlaps, a highly hydrophobic, insoluble tryptic peptide was sequenced for seven steps by the currently used manual dansyl-Edman degradation procedure, which was slightly modified for application on insoluble peptides. PRR1 coat protein contains 131 amino acids, corresponding to a molecular weight of 14534. It is highly hydrophobic, and the residues with ionizable side chains are distributed unevenly: acidic residues are absent in the middle third of the sequence, whereas a clustering of basic residues occurs between positions 44 and 62. PRR1 coat protein was compared with the coat proteins of RNA coliphages MS2 and Q beta, and the minimum mutation distance was calculated for both comparisons. It is highly probable that PRR1. Q beta and MS2 share a common ancestor. The basic region present in the three coat proteins is recognized as an essential structural feature of RNA phage coat proteins.


Biosynthesis of peptidoglycan in Pseudomonas aeruginosa. 1. The incorporation of peptidoglycan into the cell wall.

Ether-treated cells of Pseudomonas aeruginosa catalyze the formation of crosslinked peptidoglycan from the two nucleotide precursors uridinediphospho-N-acetylglucosamine and uridinediphospho-N-acetylmuramyl-L-alanyl-D-gamma-glutamyl-meso-diaminopimelyl-D-alanyl-D-alanine. The main enzymatic reactions of biosynthesis were similar to those found in Escherichia coli. Part of the reaction products were soluble in 4% sodium dodecylsulfate whereas the other part was covalently bound to the preexisting cell wall peptidoglycan sacculus. The incorporation into cell wall is carried out by a transpeptidation reaction in which the nascent peptidoglycan functions mainly as the donor and the preexisting one as acceptor. The detergent-soluble peptidoglycan is composed of partially crosslinked peptidoglycan strands as well as low-molecular-weight peptidoglycan fragments. Pulse-chase biosynthesis experiments show that the detergent-soluble peptidoglycan is an intermediate that eventually becomes covalently bound to the wall. The DD-carboxypeptidase activity of P. aeruginosa is membrane-bound and does not hydrolyse C-terminal D-alanine residues from the L-lysine-containing nucleotide-precursor analogue. An LD-carboxypeptidase was also detected in P. aeruginosa.


Biosynthesis of peptidoglycan in Pseudomonas aeruginosa. 2. Mode of action of beta-lactam antibiotics.

The intrinsic effect of various beta-lactam antibiotics on the biosynthesis of peptidoglycan of Pseudomonas aeruginosa X-48 was investigated. Most of the cephalosporins and penicillins tested already at 0.5 microgram/ml strongly inhibited (a) the incorporation of nascent peptidoglycan into the detergent-insoluble fraction (greater than 75%), (b) the formation of peptide crosslinkages (greater than 60%) and (c) the activity of the DD-carboxypeptidase and partially that of the transpeptidase (approximately 90% and approximately 40% respectively). Another group of beta-lactum drugs did not inhibit incorporation into the material insoluble in sodium dodecylsulfate, the formation of peptide crosslinkages nor transpeptidase activity. They only partially inhibited the activity of the DD-carboxypeptidase--endopeptidase system (40--50% at 0.5 microgram/ml). The results obtained differ from those of Presslitz and Ray [Antimicrob, Agents Chemother. 7, 578--581 (1975)] and show some resemblance to the effects of beta-lactams on the biosynthesis of Escherichia coli peptidoglycan.


Effect of FR 7534, a new calcium antagonist, on myocardial oxygen demand.

The actions of FR 7534, a new calcium antagonist, nitroglycerin, and dipyridamole on myocardial oxygen demand have been compared. Six anaesthetized dogs received two infusion levels of each drug which produced equivalent arterial hypotension. All three compounds reduced the tension-time index (TTI). FR 7534 and nitroglycerin but not dipyridamole produced significant decreases in myocardial oxygen consumption (MVO2). FR 7534 demonstrated oxygen-sparing actions comparable to nitroglycerin. FR 7534 may prove beneficial in alleviating myocardial ischemia.


Centrally administered neurotensin: activity in the Julou-Courvoisier muscle relaxation test in mice.

Intracisternal (IC) neurotensin (NT) produces muscle relaxation in the Julou-Courvoisier traction test, a screening procedure utilized for assessing neuroleptic drug activity. A dose-response relationship was not observed. IC administration of thyrotropin-releasing hormone (TRH) totally abolished the effects of NT. Two other peptides, substance P and bradykinin, were inactive in the traction test. These data provide further evidence for CNS effects on NT and indicate that this peptide exerts neuroleptic-like activity in a screening test for antipsychotic agents.


[Automatic rearing system of mice and transmission of pseudomonas aeruginosa].

Mice either excreting or not excreting Pseudomonas aeruginosa in feces were maintained in stainless steel mesh cages on an automated rearing apparatus with automatic water-supply nozzles and intermittently flusing metal racks. No evidence of transmission of the organisms from positive mice to negative ones was obtained during at least eight weeks.


Effect of active and passive antibacterial immunization on skin allograft survival in rabbits.

The effect of active and passive antibacterial immunization on transplantation immunity was investigated in rabbits. Immunization with dead bacterial vaccines prepared from E. coli, Proteus mirabilis and Pseudmonas aeruginosa had no effect on skin allograft survival, whereas immunization with vaccines prepared from streptococcal and staphylococcal strains shortened allograft survival. Similar results were obtained with specific antisera against individual microbial species. Skin allografts survived in rabbits treated with antisera against E. coli, Proteus mirabilis and Pseudomonas aeruginosa in the same manner as in untreated controls, whereas allograft survival was shortened with antisera against streptococci or staphylococci. The possible mechanism of the stimulatory effect of Gram-positive cocci on transplantation immunity is discussed and the possibilities of using active and passive antibacterial immunication in clinical transplantation are considered.


[Anterior pituitary function in patients with cerebrovascular disease: with special reference to growth hormone secretion in response to TRH administration (author's transl)].

HGH secretion in response to TRH was studied in patients with cerebrovascular disease in order to elucidate an influence of cerebrovascular lesions on the hypothalamus-anterior pituitary function. Blood specimens were obtained before and at the time of 10, 20, 30, 40, 60, 90 and 120 minutes after the intraveneous administration of 200 microgram of TRH in 17 patients with cerebral hemorrhage and in 8 patients with cerebral infarction. With regards to the natural fluctuations of serum HGH caused by cerebrovascular disease, the blood specimens were obrained at the time of 0, 30, 60, 90 and 120 minutes without the administration of TRH in 4 patients with cerebral hemorrhage and in 4 patients with cerebral infarction. Serum HGH was measured using the RIA method and the following results were obtained: 1) An increase in serum HGH was observed in 1 patient with cerebral hemorrhage out of 8 patients with cerebrovascular disease without TRH administration. 2) In 5 out of 17 patients with cerebral hemorrhage, an increase in serum HGH was observed following TRH administration. Three out of these 5 patients were included in the group of non-coma, and 4 out of the 5 patients were female. 3) In 2 out of 8 patients with cerebral infarction, an increase in serum HGH was observed following TRH administration.


Implantable artificial capillary unit for pancreatic islet allograft and xenograft.

Implantable artificial capillary units containing approximately 1,200 allogeneic rat islets, or approximately 3,000 xenogeneic rabbit or human islets as an implantable artificial endocrine pancreas (IAEP) were implanted in streptozotocin-induced (55 mg/kg) diabetic rats. A total of 26 rats received IAEP containing allogeneic islets. Twenty were short term experiments which lasted for 12-24 h. Four recipients survived between 1-3 days and the remaining two for 4 and 11 days respectively. Five diabetic rats received IAEP containing xenogeneic islets. The four recipients of IAEP containing rabbit islets survived up to 4 days while the recipient of IAEP containing human islets survived for 8 days. Following implantation, a decrease of plasma glucose from the initial value of 500 mg/dl to normoglycaemia and a corresponding increase in circulating levels of insulin up to 100 muU/ml were observed in the recipient animals. Furthermore the IAEPs were shown to produce a near normal plasma glucose and insulin response to an intravenous glucose tolerance test. These findings suggest the feasibility of achieving amelioration of diabetes with allogeneic or xenogeneic pancreatic islets implanted as an artificial endocrine pancreas unit in the experimental animals which has the potential of future clinical application in man.


Unusual base sequence arrangement in phage phi 29 DNA.

Susceptibility of Bacillus subtilis phage phi 29 DNA to 34 different restriction endoculceases was determined. Three enzymes, BglI, XbaI and BstEII, were found to cleave phi 29 DNA only once at specific sites. The sites of these single cleavages have been mapped. Thirteen enzymes did not cut phi 29 DNA. phi 29 HindIII DNA fragments inserted into pBR313 plasmid and propagated in Escherichia coli, were resistant to these restriction endonucleases. This result suggests that the insusceptibility is due to the absence of the nucleotide sequences on phi 29 recognized by the enzymes, and not to the presence of modified nucleotides.


Disorders in peripheral arterial system in asymptomatic elderly: plethysmographic semiology at rest, during postural, effort and pharmacological tests.

The vascular system of 30 old asymptomatic patients (average age 62.5 years) was studied by reflection plethysmography. The plethysmogram (PTG) was recorded from the forefinger of the left hand at rest, during two postural tests (+45 degrees arm-up and -45 degrees arm-down), after physical work, and during the infusion of nitroglycerin and the beta-agonist metaproterenol. 50% of the subjects presented a normal PTG under basal conditions. However, pathological PTGs were recorded after the various tests: 23.3% with the arm-up test, 6.6% with the arm-down test, 52% with the effort test, 31% with the nitroglycerin test and 73.9% with the metaproterenol test. Clearly, the effort and metaproterenol tests are pathological in the majority of cases, both tests inducing physiologically vasodilation and increase in the peripheral pulse. The arm-down test, which usually induces vasoconstriction, is almost always normal. Since the percentage of pathological responses to the nitroglycerin test is significantly smaller than that to the effort and metaproterenol tests, it is concluded that the vascular changes induced by aging are primarily functional, at least in asymptomatic subjects. Aging more negatively influences the vasodilating than the vasoconstricting ability.


[The treatment of hepatic emergencies].

Hepatic emergency means impending or manifest hepatic insufficiency. It is caused by acute and even more often by chronic liver diseases. The patient has to be hospitalized for treatment. Basic treatment is of utmost importance. It can be done in every intensive-care unit: Careful monitoring and treatment of any complication as well as reduction of toxic protein degradation products by protein restriction, bowel emptying, and enteral application of lactulose and/or neomycin. The infusion of special aliphatic amino acid solutions is still an experimental treatment. In addition to this basic treatment there are only very few indications for a special treatment: These are the prophylactic application of cimetidine in fulminant hepatitis, the hemoperfusion in intoxications by amanita or paracetamol, and the forceful hyperventilation (respiration volume 25-30 l/min) in intoxications by halogenated carbohydrates, and perhaps steroid treatment in severe alcoholic hepatitis. All heroic forms of treatment like exchange transfusion and monkey liver perfusion still have to prove their effectiveness.


[Ureter replacement by means of the umbilical vein].

Using a mixed lymphocyte culture and the 51-chromium release cytotoxicity assay, the antigenicity of umbilical vein of mice was found to be lower than the skin and the vena cava of the same embryo. In 13 dogs the middle-third of the ureter, and in 4 dogs the entire ureter, were replaced. The mean success rate was 60%. After two years, the umbilical vein showed ureter-like behavior functionally and histologically.


Thyroid hormone levels in the developing obese-hyperglycemic syndrome.

Examination of the thyroid hormone levels of C57BL/6J ob/ob mice and their lean littermates between the ages of 6 days and 22 weeks revealed that obese mice have significantly reduced hormone concentrations between 10 and 21 days of age. Thereafter, the values remained equal to, or above those or their lean littermates. The present data indicate that the hypometabolism and abnormal thermal regulation of weanling mice could be a result of diminished thyroid hormone levels, but that other mechanisms must be responsible for the persistence of these abnormalities in adult ob/ob mice.


Effects of propylthiouracil and methylmercaptoimidazol on metabolism of thyroid hormones by cultured monkey hepatocarcinoma cells.

Monkey hepatocarcinoma cell monolayer cultures (NCLP-6E) metabolized thyroxine, 3,5,3'-triiodothyronine, 3,3',5'-triiodothyronine and 3,3'-diiodothyronine by phenolic and nonphenolic ring deiodinations and sulfation of the deiodinated products, as shown in previous work with this system. The effects of the antithyroid drugs, propylthiouracil (PTU) and methylmercaptoimidazole (MMI), on these processes was investigated. PTU, at 0.1 and 1 mM, inhibited only phenolic ring deiodination. MMI at 1 mM had no effect, but 32 mM inhibited deiodination of both rings as well as sulfation. The findings suggest that the increased serum rT3 level caused by PTU in vivo is the result of decreased rT3 deiodination, in contrast to the increased rT3 production which is caused by starvation.


Relationship of iodide-induced increase in TSH response to TRH to changes in serum thyroid hormones.

Large doses of iodide (500 mg three times a day) administered to normal men for 10--12 days caused a rise in basal serum TSH and a concomitant rise in the peak TSH response to TRH. The basal and peak levels of TSH were highly correlated (p less than 0.001). However, the iodide-induced rise in the peak TSH after TRH was poorly correlated with concomitant changes in serum thyroid hormones. Serum T3 wa not lower after iodide and, while serum T4 was somewhat lower, the fall in serum T4 was unexpectedly inversely rather than directly correlated with the rise in the peak TSH response to TRH. Thus, increased TSH secretion after iodide need not always be directly correlated with decreased concentrations of circulating thyroid hormones even when large doses of iodide are used. Clinically, a patient taking iodide may have an increased TSH response in a TRH stimulation test even though there is little or no change in the serum level of T3 or T4.


Malignant schwannoma of the stomach in a patient with von Recklinghausen's disease.

A patient with von Recklinghausen's disease died with a malignant schwannoma of the stomach and was found at autopsy to have neurofibromatosis of the gastrointestinal tract, a plexiform neurofibroma of the myocardium and a phaechromocytoma. Malignancy of the gastrointestinal tract in von Recklinghausen's disease is rare, and this case highlights the difficulties in histological diagnosis of malignant nerve sheath tumours.


Diploma schools foster professionalism.

Data show that diploma schools still offer and probably will continue to offer an extremely important means of nursing education.


Is ambulatory surgery less expensive?

To determine whether an inhospital or a freestanding ambulatory surgery program can contain costs for its community, a hospital must analyze a variety of considerations.


Is medical technology safe?

The public, policymakers, and health care professionals increasingly are asking whether the risks of certain technologies outweigh their benefits.


Immunological surveillance against neoplasia: an immunological quandary.

Immunological endeavor in recent years calls for a reappraisal of the concept of immunosurveillance against neoplasia. This concept proposes an immunological policing system capable of aborting tumor growth by the recognition of "nonself" tumor associated antigens on neoplastic cells. The model is supported by evidence of tumor induction in the immunosuppressed host and the demonstration of an immune response to tumors in animals. The occurrence of tumor, regarded as a failure of immunosurveillance, is attributed to selection of neoplastic cells for immunological or other reasons or abnormal humoral or cellular antitumor immune responses. However protagonists of the postulate are faced with mounting evidence that fails to support the surveillance hypothesis. These observations include, inter alia, the monoclonality of certain tumors, the low incidence of spontaneous tumors in genetically immunodeficient mice and immunological privileged sites, and new ideas about the pathogenesis of lymphoproliferative neoplasms. However, contradictory arguments are not sufficiently substantiated to prosecute the case against surveillance conclusively. In citing highlights of the evolving quandary, both the pros and cons of immunological surveillance are presented here.


A two-year-old patient with an atypical expression of GM1-beta-galactosidase deficiency: biochemical, immunological, and cell genetic studies.

Cultured skin fibroblasts from a 2-year-old boy with an atypical form of beta-galactosidase deficiency have been studied. With the artificial substrate 4-methylumbelliferyl-beta-D-galactopyranoside, 5--15% residual activity was found in fibroblasts from this patient. Most of this activity was in the monomeric A form of the enzyme, very little in the multimeric B form. Km value, pH profile, and heat lability of the mutant enzyme were similar to those of beta-galactosidase from control fibroblasts. Immunological studies showed that the mutant enzyme cross-reacted with an antiserum raised against human liver beta-galactosidase, but the catalytic activity per unit antigenic activity was lower than normal. It was demonstrated by somatic cell hybridization that the gene mutation in this patient is different from that in patients with type 1 or type 2 GM1-gangliosidosis. No genetic complementation was found after fusion of fibroblasts from this patient with those from two other clinical variants of GM1-gangliosidosis formerly designated type 3 and adult type 4.


[The behavior of bioelements during long-term parenteral feeding].

26 adult patients were investigated for 26--35 postoperative days while being fed parenterally. Serum levels and urine excretion of zinc, magnesium, calcium and phosphate were measured. From the results it can be assumed that only phosphate substitution is necessary during parenteral long-term nutrition.


[Comparative studies on perioperative infusion therapy in children].

Various infusion solutions--containing different amounts of sodium, potassium, chloride and carbohydrates--were compared within two groups of infants and children from six months to fourteen years of age. Among various parameters particularly blood electrolytes and urinary electrolytes were taken as important parameters to recommend certain types of the basic infusion solutions, to be used for the perioperative period in pediatric surgical cases.


[Postoperative parenteral feeding with a complete solution].

Post-aggression metabolism leads even after moderately severe surgical interventions to postoperative catabolic derangement if there is inadequate replacement of carbohydrates, amino-acids and electrolytes. An investigation was carried out in 12 patients to see whether, and to what degree, the parenteral administration of a solution for total parenteral nutrition helped to bridge the postoperative nutrient deficiency. It is shown that with this form of nutrition and a complication-free postoperative course there was no imbalance of the nutritive metabolism.


[Use of maltose and a mixture of maltose, fructose and xylitol in parenteral feeding].

Maltose or maltose in combination with fructose and xylitol was administered intravenously to eight healthy male subjects. Constant maltose levels could not be attained in the blood at an infusion rate of 0.125 g maltose/kg body-weight and hour. Maximal concentrations of maltose were found at the end of the infusion period. 8.6 +/- 1.2% of the administered radioactivity was excreted into urine within 8 hours. Regarding the enzymatically determined maltose and glucose, the maltose balance was more favorable with the loss of only 3--3.5% carbohydrates into urine. The highest oxidation rate of the administered maltose was 1.5 g maltose/human volunteer and hour. During the experimental period of 8 hours 7.4 g maltose, corresponding to 37% of the applied dosis of the disaccharid, has been oxidized to and excreted as 14CO2. Xylitol and fructose did not effect utilisation and balance of maltose. Only the urinary excretion of glucose was higher when the combined solution was applied. At a limited infusion rate (0.125 g maltose/kg body-weight and hour) maltose or the combined solution maltose--xylitol--fructose may be recommended for parenteral nutrition.


[Supply of amino acids only in the early postoperative phase. Preliminary report].

The effect on protein metabolism was studied in patients, who received 4 different types of peripheral-venous infusions in the early postoperative period. Group 1 (control): Electrolytes + vitamins; group 4: 1 g/kg b. w. amino-acids alone; group 6: 1 g/kg b. w. amino-acid plus 2 g/kg b. w. carbohydrates, and group 7: 1 g/kg b. w. amino-acids plus 5.5 g/kg b. w. carbohydrates. Compared to the control-group in all patients supplemented with amino-acids a significantly improved 24 h-nitrogen balance was observed. Regarding the cumulative nitrogen balance (4 days), it was noticed that the balance was more negative in patients given amino-acids alone than in patients, who received carbohydrates additionally. At no time serum protein levels (albumin, transferrin, prealbumin and retinol-binding protein) were found to be significantly improved by the addition of amino-acids alone. Therefore, our study indicates that there is no reason to leave the proven concept of peripheral-venous feeding with amino-acids plus carbohydrates to the favor of sole amino-acid infusions in the early postoperative period.


Clinical-pharmacological aspects, application and effectiveness of total parenteral nutrition in surgical patients.

The term "total parenteral nutrition" (TPN) refers to the maintenance of an adequate nutritional status, normal body weight and positive nitrogen balance solely by intravenous means. It requires solutions providing calories, amino acids and other nutrients in amounts much greater than those indicated for maintenance of normal body weight. Nutrient solutions have been studied, selected and prepared in our Hospital Pharmacological Service utilizing a sterile closed system, which allows large-volume filtering, sterilizing and bottling devices. For maintenance of weight gain in adults, a basic formula is employed, which provides 1,100 Kcal/1 with pure crystalline amino acids mixed with 50% anhydrous dextrose in water in a ratio of 5.8:1 (160 Kcal:1 g nitrogen). Minerals and vitamins are added to the base solution prior to use and may be increased or decreased by simple addition or omission depending on the patient's condition. This paper is based on 192 surgical patients who received TPN and have been followed in strict cooperation between the Hospital Pharmacological Service and the Surgical Department. The patients, ranging from 23 to 79 years of age, with life threatening diseases and unable to maintain adequate nutrition by the oral route, received TPN through a central catheter inserted via subclavian puncture (146 cases) or through a surgically created internal A-V fistula (46 cases). The condition of the patients generally improved within a few days after starting TPN; and weight gain, wound healing, general improvement and a shorter period of hospitalization were observed. TPN could be efficiently combined with oncologic treatment, and a significant improvement of the patients' performance status and decrease of toxic side-effects due to chemotherapeutic agents were observed. TPN has been successfully applied also in patients with fistulas of the alimentary tract obtaining spontaneous closure and in patients with ulcerative colitis, showing its beneficial effect in allowing complete bowel rest for healing. No major complications or deaths could be attributed to TPN or to the route of administration.


Penetration through the gram-negative cell wall: a co-determinant of the efficacy of beta-lactam antibiotics.

Resistance of gram-negative bacteria to beta-lactam antibiotics is based mainly on two mechanisms: hydrolysis by beta-lactamases and exclusion of the antibiotics from their target sites in the inner membrane. This article describes the use of Pseudomonas aeruginosa K 799/WT and a mutant of this strain (K 799/61) to assess the role of the outer membrane as a permeability barrier to penicillins and cephalosporins. The data confirm the importance of good penetration for a beta-lactam to be active against Pseudomonas. The second part illustrates the interplay of beta-lactamases and the outer membrane in the resistance of Escherichia coli to beta-lactams. A method to determine membrane permeability parameters parameters is given. The results support the idea that only a combined consideration of inactivating enzymes and penetration barriers can lead to a better understanding of the efficiency of the defence mechanisms which gram-negative bacteria can invoke against beta-lactam antibiotics.


The groove of the mandibular neck as a risk factor in vertical osteotomy of the ramus of the mandible.

Surgical damage of anatomical structures within the mandibular canal represents one of the risks of dysgnathia operations performed on the ramus of the mandible. The paper presents a review of the various measurements of anatomical landmarks available in the pertinent literature. The occurrence of the sulcus coli mandibulae, as well as the depth of the groove have been studied. In some cases the groove has to be considered as a surgical risk factor. On the basis of the examination, attention is called to the individual variations and to the necessity of a cautious surgical technique.


Treatment of oral hemangiomas with local sclerosing agents.

Twenty-four cases of hemangiomas of various sizes in the oral cavity and lips were treated with local injections of a sclerosing agent (3% Sotradecol - sodium tetradecyl sulfate). No side effects were noticed and the lesions disappeared without scarring. The article discusses the practical aspects of this therapy.


Influence of Alvogyl on the healing of extraction wound in man.

Eight volunteer patients free of any systamic disease underwent extraction of two teeth on the same day. One extraction socket was packed with Alvogyl, the other served as a control wound. Biopsies for histological assessment were taken from both sockets 1 and 2 weeks after extraction. Histological study revealed the normal sequence of wound healing in the sockets allowed to heal spontaneously. On the other hand, the signs of retarded wound healing (acute inflammatory infiltrate, persistent granulation tissue, failure to form connective tissue scar and frequent foreign bodies/foreign body giant cells) were encountered in the sockets packed with Alvogyl. On the basis of these preliminary results, it seems warranted to state that the systematic use of Alvogyl as a preventive and curative treatment for postextraction alveolar infections cannot be advocated.


Postoperative swelling and trismus after mandibular third molar removal with the lingual split bone technique.

Several factors have been suggested as possible causes for the development of the swelling and trismus which occur after the surgical removal of impacted third molar teeth. In this investigation, measurements of swelling and trismus were made before and after lower third molar removal by the lingual split technique, in a series of 53 patients. No single cause of these surgical sequelae was identified and the duration and pattern of both swelling and trismus were recorded.


Collection of cytological specimens with a cell aspirator with reference to the wooden spatula method. II. Comparative quantitative and qualitative studies.

This article describes the results of comparative quantitative and qualitative examinations of specimens obtained by testing 65 individuals by means of a cell aspirator and a wooden spatula. The studies were performed on a clinically normal mucosa as well as on different mucosal lesions. A statistical analysis of the cells obtained showed that the cell aspirator quantitatively produced significantly better results both with regard to the total number of cells obtained and to the number of epithelial cells in the different stages of maturity. The number of cells obtained turned out to be dependent on surface structure and the size of the lesion, however. Qualitatively the cell aspirator collected epithelial cells which were better preserved morphologically, but also a larger number of compact cell clusters. The number of erythrocytes was low. The structure of these blood cells was well preserved, indicating bleeding arising in connection with the taking of the specimens. The number of leucocytes was larger in the aspirated material. The degrees of fixation and staining were similar. The authors make recommendations for using the cell aspirator in such a manner as to obtain the best qualitative results.


Effect of different levels of dietary zinc on development of chemically induced oral cancer in rats.

Four-week-old female rats distributed among three experimental groups were fed a 0.23 (zinc-deficient), 0.77 (zinc-adequate) or 3.06 (zinc-supplemented) mmol zinc/kg diet and oral cancer was induced by repeated application of the water-soluble carcinogen 4-nitroquinoline N-oxide (4NQO) to the palatal mucosa. The clinical development of oral cancer differed significantly among the three groups (P less than 0.05). In rats fed the zinc-supplemented diet, the macroscopic oral cancer appeared early and the survival time was shorter than in either of the other groups. It was concluded that a zinc-supplemented diet accelerates, and a zinc-deficient diet retards the development of chemically induced oral cancer in rats.


Basal cell nevus syndrome. A case report.

An 11-year-old boy with multiple dentigerous cysts in the maxilla and mandible is described. Other findings seen in the face, plantar skin, skeletal system and oral cavity indicated the lesions to be due to the basal cell nevus syndrome. This was further confirmed by the presence of similar abnormalities in his father and brother.


Oral bony lesion in a patient with medical history of hyperparathyroidism.

Primary hyperparathyroidism is an uncommon disease in which the amount of parathyroid hormone circulating in the bloodstream is increased. Therefore, bony changes are often found in the prolonged or severe cases of hyperparathyroidism. This study describes and discusses the relationship between cystical radiolucent lesion in the left mandible of 29-year-old Japanese male and hyperparathyroidism which has the feature of bony changes.


Clinical evaluation of Warthin's tumor. An analysis of 43 cases.

Forty-three patients with Warthin's tumor (26 males and 17 females) were analyzed and the following results were obtained: All tumors were related to the parotid gland with no bilateral involvements. The ages of the patients were 20-84 years old and the average age was about 62. The interval between tumor recognition and first examination was 1 week to 16 years with the average interval of 20-22 months. The method of treatment was complete enucleation. However, radiation therapy was also done in three cases, two of which were irradiated after operation. The other received palliative radiation therapy because of old age and poor general condition. Recurrence was observed in two cases in the course of the follow up, but the examination at the end of June 1977 revealed neither recurrence nor metastasis in the cases followed. A rare case of synchronous occurrence of warthin's tumor and another histological tumor was included in this series and similar cases are reviewed briefly in the literature.


The effects of topical pilocarpine on the morphology of the outflow apparatus of the baboon (Papio cynocephalus).

The morphology of the outflow system of the baboon (Papio cynocephalus) was studied by light microscopy and transmission electron microscopy in tissue from eyes treated with topical pilocarpine. In one group of five animals the intraocular pressure (IOP) was maintained at 18 mm /g in both the experimental and the control eye of each baboon for the duration of the experiment. The endothelial meshwork was more distended, and there was a significantly greater number of giant vacuoles in the lining endothelium of Schlemm's canal in the pilocarpine-treated eyes than in the corresponding controls. However, in another series of animals, IOP was not maintained, and the differences between the treated and untreated eyes was much less striking. There appeared to be little evidence that the increase in vacuolation was produced by the direct action of pilocarpine on the endothelium of Schlemm's canal; more likely, it was a consequence of an increased passage of fluid through the drainage system. In addition, we have been able to show that (1) the mode of primary fixation can produce partial reversal of the pilocarpine effect and (2) the mechanical manipulation which is required to determine outflow facility has a deleterious effect on the morphology of the outflow tissues.


A light microscopic study of the effects of testicular hyaluronidase on the outflow system of a baboon (Papio cynocephalus).

An attempt was made to investigate the effects of hyaluronidase on the morphology of the baboon outflow system. The eyes of six adult baboons provided the material for this histological study. For each animal the IOP in both eyes was maintained at 18 mm Hg, and 150 IU of testicular hyaluronidase were introduced into one eye and a control solution into the other. It was observed by light microscopy that the outer meshwork was more distended in the hyaluronidase-treated outflow system than in the corresponding controls. Further, it was found from a quantitative analysis that the incidence of giant vacuoles in the endothelium of Schlemm's canal was greater in the experimental than the control eye of each animal. The difference between the vacuole counts in the experimental and control eyes was significant in five of the six animals. These preliminary findings provide morphological evidence which indicates that there is a hyaluronidase-sensitive barrier to aqueous outflow in the baboon drainage system.


Rubella virus maturation and production in two host cell systems.

When inoculated at the same MOI, Vero cells released a larger amount of infectious rubella virus into the culture medium than did BHK21 cells, However, BHK21 cells (in monolayer or in suspension) produced more intracellular infectious virus than Vero cells when tested 24 h after infection. Maturation of the virus in BHK21 cells occurred at the plasma membrane and, in a larger quantity, in the cytoplasm (Golgi apparatus and vacuoles). Viral particles consisted of an electron-dense core (32 nm) surrounded by a capsid and enveloped by a single membrane (8-10 nm). Aberrant forms (elongated and twisted) in the vacuole and double virions in the plasma membrane were observed as early as 65 h after infection.


Antibodies in human sera reacting with an insect pathogenic virus.

Precipitating antibodies to an insect pathogenic RNA virus of Darna trima from East Malaysia have been found in a small percentage of human sera from several different groups of persons in West Malaysia and the United Kingdom. No associated illness was identified. The results suggest that an antigenically related virus or viruses are present in the environment that may be associated with symptomless or inapparent infections in man.


Effect of human serum thymic factor on immature T lymphocytes in acute leukemia and other hematologic malignancies.

In 37 patients with acute leukemia and in 13 patients with other hematologic malignancies, E-rosette formation of peripheral blood mononuclear cells, before and after incubation with human serum thymic factor, was studied. This assay showed that incubation with thymic factor caused a clear-cut increase of E-rosette-forming cells in 8 of the 37 acute leukemia patients. Among the 13 patients with other hematologic malignancies, a similar effect was observed in two with pediatric Hodgkin's disease and in one adult with Waldenström's macroglobulinemia. These results support the contention that a significant percentage of the so-called "null" cells circulating in the blood of the above-mentioned patients were, in fact, immature T cells (T0 lymphocytes) that were shifted to a more mature stage (T1) by thymic factor.


Leukocyte and differential counts after cardiac surgery in children.

In order to obtain information about the extent and duration of the changes in the leukocyte and differential counts following cardiac surgery done with and without extracorporeal circulation in the absence of any infection, 28 children were studied serially during the first ten postoperative days. In patients operated without extracorporeal circulation, normal values are reached after 24--48 h; in patients operated with extracorporeal circulation, normal values are reached only after 4--8 days, depending on the length of extracorporeal circulation.


Histochemical localization of glycosidases in dog epididymis.

The histochemical localization of five glycosidases was studied in the epididymis of mature dogs. beta-Galactosidase showed a distinct to strong reaction in the epithelium of the ductuli efferentes and throughout the whole length of the ductus epididymidis. beta-N-Acetylglucosaminidase reactivity was weak in the initial segment, but increased significantly in the middle and terminal segment. The maximum beta-glucuronidase activity was found in the ductuli efferentes and in the initial segment. The alpha-mannosidase reaction was weak in all segments except the middle segment where a distinct activity was seen. With the method employed, no alpha-fucosidase activity could be detected. The physiological role of the glycosidases in the epididymis is discussed briefly.


Rapid demineralization in acidic buffers.

The demineralization of routine histological specimens in buffers of weakly ionized organic acids, unbuffered formic acid, and EDTA was investigated. The rate of demineralization was measured by a chemical method and from radiographs. Lactate-containing buffers and buffers of formic acid with its potassium salt were more rapid in effect than any other agent. Acidic buffers and unbuffered formic acid produced rapid diffuse demineralization with secondary precipitation of calcium salts. Preservation of dental enamel in such buffers resulted from the significantly slower rate of enamel demineralization than that for bone and dentine. In rapid demineralizing agents the secondary salts were quickly redissolved while in slow buffers these salts persisted. Multivalent ions such as citrate and maleate slowed the rate of demineralization, and a citrate-containing buffer was the slowest of all the agents tested. Demineralization in EDTA exhibited a different pattern with the establishment of a well-defined front of demineralization without apparent reprecipitation. EDTA attacked enamel, bone and dentine at the same rate. An attempt was made to relate the observed rates of demineralization to current theories of the demineralization process.


Amino acid sequences of the alpha and beta chains of adult hemoglobin of the brown lemur, Lemur fulvus fulvus.

Globin prepared from hemoglobin of the brown lemur (Lemur fulvus fulvus) was separated into alpha and beta chains by chromatography on a CM 52 column. The S-aminoethylated alpha and beta chains were each digested with trypsin and resulting peptides were isolated. The amino acid sequences of the tryptic peptides were established. The ordering of these peptides in the alpha and beta chains was deduced from the homology of their amino acid sequences with that of human adult hemoglobin. The primary structure of brown lemur hemoglobin thus obtained differs from that of human hemoglobin in 15 amino acids in the alpha chain and 26 in the beta chain.


Regulation of teichoic acid synthesis during phosphate limitation.

Bacillus subtilis W-23, when placed in phosphate-free medium, ceases to synthesize teichoic acid and synthesizes teichuronic acid. The enzymatic basis for the cessation of teichoic acid synthesis is the irreversible inhibition of the first membrane-bound enzyme involved in teichoic acid synthesis which catalyzes the reaction Undecapenol-P + UDP-GlcNAc leads to undecaprenol-P-P-GlcNAc + UMP.


Purine nucleoside phosphorylase cleaves the C--O bond of ribose 1-phosphate. Evidence from the 18O shift in 31P NMR.

An equilibrium mixture of highly enriched [18(O)]Pi (represents the mixture of [[18(O)4]Pi, [[18(O)3]Pi, [18(O)2]Pi as represented in the figures, unless otherwise specified), alpha-D-ribose 1-[16(O)]phosphate, and hypoxanthine plus inosine was equilibrated with calf spleen purine-nucleoside phosphorylase (EC 2.4.2.1). The 31P NMR spectrum clearly indicated the formation of alpha-D-ribose 1-[18(O)4]-phosphate and of [16(O)]Pi. Incubation for the same time span in the absence of alpha-D-ribose 1-phosphate left the [18(O)4]Pi isotopic distribution unchanged. The results clearly demonstrated that the C--O bond of alpha-D-ribose 1-phosphate is cleaved in the enzymatic reaction. It is unlikely that the enzyme catalyzes the exchange of oxygen between Pi and H2O. Several possible mechanistic pathways are ruled out by the results, which demand attack by a phosphate oxygen at the anomeric C-1' atom.


Metabolism of thyrotropin releasing hormone in brain extracts. Isolation and characterization of an imidopeptidase for histidylprolineamide.

An extract of porcine brain acetone powder incubated with thyrotropin-releasing hormone (TRH; pGlu-His-ProNH2) produces acid TRH (pGlu-His-Pro), histidine, and prolineamide. Fractionation of the brain extract by DEAE-cellulose chromatography produces three protein fractions which metabolize TRH. The activity of these fractions was characterized using TRH with a 3H-label on the histidine or proline as well as [His-3H]His-ProNH2. Fraction I contains pyroglutamate aminopeptidase and Fraction II contains TRH deamidase. Fraction III was found to contain a previously unrecognized enzyme which cleaves His-ProNH2 to histidine and proline. The histidylprolineamide imidopeptidase has been characterized. A competition study using a variety of compounds containing histidine or proline suggests that the best substrates for the imidopeptidase contain a free alpha-amino group on histidine and a blocked carboxyl group on proline, as is found in His-ProNH2. A survey of a variety of polypeptide hormones indicates that many of them inhibit the imidopeptidase activity. A kinetic study of the inhibition of the enzyme by adrenocorticotropic hormone (1-24) shows that the inhibition by polypeptide hormones is noncompetitive. We hypothesize that pituitary hormones may stimulate the production of (cyclo)-His-Pro by inhibiting alternate routes of TRH metabolism.


Microtubules and the regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase.

Cultured C-6 glial cells were utilized to evaluate the effect of antimicrotubular drugs on 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase and cholesterol synthesis. Colchicine, Colcemid, and vinblastine (1.0 muM) caused a marked reduction in HMG-CoA reductase activity and, as a consequence, the rate of cholesterol synthesis in these cells. No effect was observed with lumicolchicine, a mixture of colchicine isomers with no effect on microtubules. The effect of colchicine was apparent within 1 h after addition to the culture medium, and, after 6 h, HMG-CoA reductase activity in treated cells was only approximately 15 to 30% of that in untreated cells. Reductase activity was very sensitive to the concentration of drug added, i.e. cells treated with just 0.1 muM colchicine for 6 h exhibited a 50% lower enzymatic activity than did untreated cells. The lack of a generalized, nonspecific toxic effect on the cells was indicated by the finding of no change in the activities of fatty acid synthetase and NADPH-cytochrome c reductase and the rate of total protein synthesis in cells treated with colchicine (1 muM) for 6 h. A close temporal and quantitative correlation was observed between the effects of colchicine on HMG-CoA reductase and on a parameter of microtubular function, i.e. maintenance of glial cell shape. The data suggest that microtubules are involved in the regulation of HMG-CoA reductase and cholesterol synthesis in C-6 glial cells.


Role of hemoglobin in proton transfer to the active site of carbonic anhydrase.

The binding of bovine oxyhemoglobin to bovine carbonic anhydrase with a dissociation constant between 10(-5) and 10(-7) M has been determined by countercurrent distribution using aqueous, biphasic polymer systems. This result provides an explanation for the very efficient proton transfer between hemoglobin and carbonic anhydrase, a transfer which enhances the catalytic activity of carbonic anhydrase as measured by 18O exchange between bicarbonate and water at chemical equilibrium (Silverman, D. N., Tu, C. K., and Wynns, G. C. (1978) J. Biol. Chem, 253, 2563-2567). Two rate constants describing 18O exchange activity of carbonic anhydrase at pH 7.5 show saturation behavior when plotted against hemoglobin concentration consistent with a dissociation constant of 2.5 X 10(-6) M between bovine hemoglobin and carbonic anhydrase. Interpretation of these rate constants in terms of a two-step model for 18O exchange indicates that hemoglobin enhances the rate of exchange from carbonic anhydrase of water containing the oxygen abstracted from bicarbonate, but does not affect the catalytic interconversion of CO2 and HCO3- at chemical equilibrium.


Primary structure of a human IgA1 immunoglobulin. I. Isolation, composition, and amino acid sequence of the chymotryptic peptides.

As the initial phase of the determination of the complete covalent structure of a human immunoglobulin A, 52 chymotryptic peptides, ranging in length from 2 to 37 residues, were isolated and characterized from the reduced and carboxymethylated alpha1 heavy chain of the myeloma IgA protein Bur. The peptides were subjected to sequence analysis by the dansylation technique, manual and automatic Edman degradation, and carboxypeptidase digestion. The results, in conjunction with the data on the tryptic and thermolysin peptides and the cyanogen bromide fragments reported in the accompanying papers, established the complete primary structure of a human IgA chain.


Primary structure of a human IgA1 immunoglobulin. IV. Streptococcal IgA1 protease, digestion, Fab and Fc fragments, and the complete amino acid sequence of the alpha 1 heavy chain.

In order to establish the complete amino acid sequence of the human IgA alpha1 chain Bur, IgA1 protease from Streptococcus sanguis was employed to generate Fabalpha and Fcalpha fragments in the final stage of this investigation. Cyanogen bromide cleavage of the Fabalpha fragment followed by reduction and aminoethylation produced the Fd' fragment (residues 84 to 227); this contains part of the variable region (VR), the whole first constant domain (Calpha1), and part of the hinge region of this heavy chain. The tryptic peptides of the Fd' fragment were isolated, characterized, and sequenced. The results together with the data in the preceding papers on chymotryptic, tryptic, and thermolysin peptides permitted the complete amino acid sequence of the human IgA alpha1 chain to be established.


Retrograde alteration of 6-phosphogluconate dehydrogenase in axotomized superior cervical ganglia of the rat.

Mechanisms underlying increased activity of 6-phosphogluconate dehydrogenase (6-phospho-D-gluconate: NADP oxidoreductase [decarboxylating] EC 1.1.1.44) in axotomized rat superior cervical ganglia were explored using a highly sensitive micro-immunochemical assay employing antibodies raised in rabbits against the purified enzyme. 6-Phosphogluconate dehydrogenase was purified from rat brain more than 1700-fold by salt fractionation, anion exchange, and immunoaffinity chromatography. The purified enzyme consisted of identical subunits having molecular weights of about 48,800 which could aggregate to catalytically active isomers of various sizes; however, only one form of the enzyme was detected in freshly prepared homogenates of rat neural tissue. Physical and immunological properties of the enzyme from rat brain were similar to those from superior cervical ganglia and liver. Augmented 6-phosphogluconate dehydrogenase activity noted in superior cervical ganglia 2 days after transection of major postganglionic nerve trunks was accompanied by a parallel increase in immunoreactive protein. Michaelis constants of the enzyme were the same in control and axotomized ganglia, and the presence of activators and inhibitors was not detected. It is concluded that increases in 6-phosphogluconate dehydrogenase subsequent to axotomy can be accounted for entirely by an increase in the steady state concentration of this protein.


Isolation and characterization of a guanine insertion enzyme, a specific tRNA transglycosylase, from Escherichia coli.

A guanine insertion enzyme (tRNA transglycosylase) was purified to a homogeneous state from Escherichia coli B by ammonium sulfate fractionation and DEAE-cellulose, DEAE-Sephadex A-50, phosphocellulose, and Sephadex G-200 column chromatographies. The molecular weight of the enzyme, which appeared to be a single polypeptide, was 4.6 X 10(4) by sodium dodecyl sulfate gel electrophoresis. The enzyme catalyzes exchange of guanine with guanine located in the first position of the anticodon of tRNATyr, tRNAHis, tRNAAsn, and tRNAAsp, but unlike the enzymes isolated from rabbit reticulocytes and Ehrlich ascites tumor cells it does not catalyze the exchange of guanine with queuine (7-(3,4-trans-4,5-cis-dihydroxy-1-cyclopenten-3-ylaminomethyl)-7-deazaguanine) present in these tRNAs. The pH optimum of the reaction was 7.0, and the pH1 value was 4.6 to 4.8. The reaction required Mg2+ ion. 7-Methylguanine inhibited guanine insertion, but the other purine analogues tested were not inhibitory and could not replace guanine.20


Overall pathway of mononucleosome production.

Five electrophoretically distinguishable classes of mononucleosomes (MI, MII, ...MV) are produced upon treatment of mammalian nuclear chromatin with micrococcal nuclease. These five forms differ in their initial DNA lengths, relative mass proportions, stability, contents of histone H1, and presence of certain nonhistone proteins. A new "chromatin fingerprinting" technique has been developed in order to trace nuclease-mediated interconversions between these mononucleosomes and their polynucleosomal precursors. Application of this technique, together with earlier findings from this laboratory, has made possible the elucidation of the overall pathway of nuclease cleavage of chromatin which leads to the production and interconversion of these mononucleosomes, and has permitted reconstruction of the organization of these mononucleosomes in undigested chromatin...


Biological properties of antibodies against rat adipocyte intrinsic membrane proteins. Dependence on multivalency for insulin-like activity.

Antisera from rabbits injected with rat adipocyte plasma membranes or intrinsic proteins from such membranes, obtained by a dimethylmaleic anhydride extraction step, mimicked the action of insulin on both glucose transport and lipolysis in intact adipocytes. Biological activity in both types of antisera was mediated by immunoglobulin binding to one or more intrinsic proteins of the adipocyte plasma membrane since fat cells were unresponsive to all antisera absorbed with dimethylmaleic anhydride-extracted membranes. Acid treatment of immunoprecipitates released antibodies which activated glucose uptake and reacted with solubilized adipocyte membranes on immunodiffusion plates. The biologically active immunoglobulin preparations failed to form immunoprecipitin lines when tested against membranes from brain, liver, lung, muscle, kidney, and spleen. Insulin-sensitive glucose uptake in rat soleus muscle did not respond to the antisera. The antibodies activated hexose uptake into fat cells and reacted with solubilized adipocyte membranes on immunodiffusion plates when rat or mouse adipocytes were studied, but not when monkey fat cells were used. The anti-membrane antibody preparations readily activated hexose uptake in trypsinized fat cells which had lost the capacity to bind or respond to insulin. These data are consistent with the concept previously proposed (Pillion, D.J., and Czech, M.P. (1978) J. Biol. Chem. 253, 3761-3764) that the anti-membrane immunoglobulins do not interact with the insulin binding site of the insulin receptor. Monovalent Fab fragments of the biologically active antisera, prepared by papain digestion of the native anti-membrane immunoglobulins, were ineffective in enhancing glucose uptake in adipocytes. However, biological activity of the anti-membrane Fab fragments was restored by the addition of goat anti-rabbit Fab antisera to cells treated with the Fab fraction. Anti-rabbit Fab antisera alone or in combination with Fab fragments prepared from control rabbit sera exhibited no biological activity. These results demonstrate that the ability of anti-membrane antisera to mimic the biological activity of insulin on isolated fat cells is critically dependent on immunoglobulin binding to one or more intrinsic plasma membrane proteins and the multivalent nature of immunoglobulin structure.