The effect of phenazine methosulfate-ascorbate on bacterial active transport and adenosine triphosphate formation: inhibition of Pseudomonas aeruginosa and stimulation of Escherichia coli.

The artificial electron-donor system, phenazine methosulfate (PMS) ascorbate, inhibited active transport of glucose by Pseudomonas aeruginosa irrespective of whether the incubation systems were in air, flushed with oxygen, or gassed with nitrogen under anaerobic denitrifying conditions. Active transport of glucose by P. aeruginosa was also inhibited by reduced 5-N-methyl-phenazonium-3-sulfonate, a membrane-impermeable electron donor. PMS-ascorbate caused rapid depletion of intracellular adenosine triphosphate (ATP) when added to respiring cell suspensions of P. aeruginosa either in the presence or absence of glucose or succinate as oxidizable energy sources. In contrast, under identical conditions, Escherichia coli formed ATP with PMS-ascorbate as the sole oxidizable energy source and ATP formation continued when glucose or succinate was present in addition to PMS-ascorbate in the incubation system.


Induction of beta-galactosidase in Streptomyces violaceus.

Synthesis of beta-galactosidase by Streptomyces violaceus was induced by D-galactose and L-arabinose, and to a lesser extent by lactose, D-arabinose, and methyl-beta-D-galactopyranoside. The synthesis of the enzyme was linear and started to increase 2--3 h after induction by galactose, reaching a maximum after 5--7 h. The highest level of specific activity was observed in 2% galactose, with an increase of 45 times over the basal level in glycerol. Isopropyl-beta-D-thiogalactopyranoside (IPTG) and methyl-beta-D-thiogalactopyranoside (TMG) inhibited induction by D-galactose, but did not influence enzymatic activity. Cellular extracts hydrolyzed O-nitrophenyl-beta-D-galactopyranoside, but did not significantly hydrolyze lactose, melibiose, p-nitrophenyl-alpha-D-galactopyranoside, p-nitrophenyl-beta-D-fucoside, or p-nitrophenyl-beta-D-glucopyranoside. Rifampicin and chloramphenicol inhibited beta-galactosidase synthesis in non-preinduced and in preinduced cells. The inhibition by chloramphenicol was reversible.


An agar--gel immunodiffusion test for detection of Brucella antibodies in human serum.

A comparison was made of results obtained with a Brucella agar--gel immunodiffusion (AGID) test and the standard tube-agglutination test on 612 human sera. Agreement between the tests was 97% when the titer was 1:160 or higher. Of 448 sera that showed no agglutination titer, 447 were negative with the AGID test. Results of the AGID test were also compared to those obtained with the 2-mercaptoethanol (2-ME) agglutination test on 148 sera that demonstrated a standard tube-agglutination titer of 1:20 or higher. All sera with a 2-ME-agglutination titer of 1:40 or higher were positive with the AGID test. Of 123 sera that showed no 2-ME-agglutination titer, 21 were positive with the AGID test. Two of these 21 sera were obtained from patients with bacteriologically proven brucellosis, and eight were from abattoir employees with suspected but not bacteriologically proven brucellosis.


Prospective randomized trial of combined oncological therapy for gastric carcinoma.

One hundred forty-two patients with all stages of gastric carcinoma were prospectively stratified into two divisions according to T.N.M. stage following, but irrespective of the type of surgical procedure. Division I (T1-3, N1-2, M0) was randomized into a control group, and a treatment group who received 2000 rad in 8 fractions over 10 days with intravenous 5 Fluorouracil (5 F.U.) at a dose of 500 mg daily x 4 days preirradiation and then 12.5 mg/kg daily for 5 days every 28 days for six courses. Division II (T4 or M1) was randomized into three groups; a control group, a group who received radiotherapy and 5 F.U. in the same schedule as division one and a group who received Thiotepa 45 mg intravenously daily for three days and then every 28 days for 6 months. Four and one-half years after commencement of the trial 86% of the patients had died. There was no difference in survival rate between the treatment and control groups, (p greater than 0.5) in Division I or II. Survival appeared to correlate with the T.N.M. stage of disease and not therapy. Blind assessment of the quality of life showed no difference between the treatment groups and the controls. In the dose schedules used, this form of oncological therapy had no effect on survival or quality of life in patients with gastric carcinoma.


Treatment of acute leukemia in protected environment units.

Thirty evaluable patients with acute leukemia (AL), aged 14 to 48-year-old received remission induction chemotherapy on a protected environment-prophylactic antibiotic program. Twenty-seven (90%) of these patients achieved complete remission and 17 remained in complete remission for 1 to 22 months. Although these patients spent 36% of their time with neutrophil counts less than 100/mm3, they spent only 20% of their time with fever. Major infection was present during only 7% of the days when neutrophil count was less than 100/mm3. No patient died of an infectious complication during remission induction therapy.


Karyotypic abnormalities and clinical aspects of patients with multiple myeloma and related paraproteinemic disorders.

Karyotypic abnormalities were detected in the malignant cells of 6 of 18 patients with multiple myeloma (MM). Six patients with benign monoclonal gammopathy, one with amyloidosis of immunoglobulin origin, and two with Waldenström's macroglobulinemia had normal karyotypes. All six MM patients with aneuploidy were in a group of 10 patients in an accelerated or relapse phase of their disease and four had high serum paraprotein levels (7.92, 6.24, 6.80, and 4.24 g/dl, respectively) when their abnormal karyotypes were detected. Five of the 6 MM patients with aneuploidy had received prior chemotherapy. Aneuploidy was not observed in 8 stable MM patients. Abnormalities of chromosome 14 were present in all 6 patients, with a 14q+ marker in 5 and loss of No. 14 in 1. A translocation between Nos. 11 and 14 was found in aneuploid cells of 2 patients who had plasma cell leukemia (PCL). However, the break point in the long arm of No. 11 differed in the 2 patients. A gain of Nos. 5, 9, and 11 was seen in 3 patients, a gain of No 1 in 2, and rearrangements of No. 1 in 5 MM patients, including all 4 who had a 14q+ marker chromosome initially. A deletion of chromosome 6 at band q25 was detected in 2 MM patients and a pericentric inversion of No. 6 (6p21 to 6q13) was seen in the patient with PCL. Three of 4 MM patients had a nonrandom loss of one chromosome 8. Two other MM patients, who were treated with melphalan and prednisone, developed acute nonlymphocytic leukemia (ANLL) 2+ and 4+ years after the diagnosis of MM. Marrow cells of one patient showed a 5q- chromosome and a constitutional translocation involving Nos. 13 and 14 during the preleukemic stage; during the leukemic phase, the karyotype evolved to 50 chromosomes including extra chromosomes 1, 6, 8, 10, and 21 and a missing 7, in addition to the originally detected 5q- and the 13/14 translocation. The peripheral blood from the other patient was hypodiploid, with a missing chromosome 7 and a translocation between 3q and 9p. These patterns of chromosome change resemble those of ANLL rather than MM and are similar to the changes seen in ANLL after treated malignant lymphoma.


Combination of radiotherapy and razoxane (ICRF 159) for chondrosarcoma.

Eight patients with 12 chondrosarcomas were treated with radiation and razoxane (ICRF 159). Two tumors in 1 patient progressed unequivocally, 3 tumors in 3 patients showed no change, and 7 tumors in 5 patients had complete or partial (more than 50%) regressions. At least 2 complete regressions have responded for more than 2 1/2 years at the present time.


Combined modality therapy of hepatic metastasis. Northern California Oncology Group Pilot Study.

Twenty-two patients with adenocarcinoma metastatic to the liver were treated with rapid fractionation whole-liver irradiation (1350-2100 rads in 300-rad fractions) with simultaneous intrahepatic 5-fluorouracil (10-15 mg/kg/day) and intrahepatic Adriamycin 2.5-10 mg/m2/day) as part of a Phase I-II study. Of the 21 patients who completed therapy, 19 had colorectal carcinoma and 2 had metastatic adenocarcinoma of unknown origin. Objective response was judged by measurement of liver size, evaluation of liver function tests, and by liver scan or CAT scan of the liver. Ten of the 21 evaluable patients responded, yielding an overall response rate of 47.6%. The response rate in patients with colorectal carcinoma was 55% (10/19). At this time, median duration of response is 14+ weeks and median survival from onset of therapy is 15+ weeks. Hematologic and gastrointestinal toxicity were tolerable. No hepatic toxicity was documented. This combined modality therapy was found to be a safe effective method for the palliation of liver metastasis.


Chronic myelomonocytic leukemia with paraproteinemia but no detectable plasmacytosis: a detailed cytological and immunological study.

A patient with chronic myelomonocytic leukemia with IgG K paraproteinemia, but no detectable plasmacytosis, is described. The patient was entering a blastic phase at the time of the most detailed studies. Cytological, cytochemical, and ultrastructural studies revealed a mixed myeloid proliferation with granulocytic forms predominating over monocytic elements. A variety of ultrastructural abnormalities, including defective granulation, was observed but no cells with highly developed rough endoplasmic reticulum were observed. Immunological marker studies showed that the mature myeloid cells possessed receptors for the Fc of IgG and weakly expressed the Ia-like P29/34 antigen. The mature myeloid cells also expressed both surface and intracytoplasmic Ig restricted to IgG K, and this IgG K persisted after 4 weeks in culture. A reverse plaque assay showed that the myeloid cells were capable of releasing IgG K in vitro, but studies involving the incorporation of radio-labeled amino acids showed no detectable Ig production by the myeloid cells. The possible interpretations of these data are discussed in some detail in relation to previous reports of paraproteinemia in myeloid proliferative disorders.


Excision of DNA damage arising from chemicals of different carcinogenic potencies.

Primary cultures of mouse embryo cells are more efficient in excising DNA-carcinogen adducts resulting from exposure to either 7-bromomethylbenz-[alpha]anthracene or the more carcinogen 7-bromomethyl-12-methylbenz[alpha]anthracene than are mouse L 929 cell suspension cultures. However, within each of these systems, the excisabilities of the adducts formed by either bromo-compound are similar, so differences in carcinogenic potency of the compounds cannot be attributed to differences in the excisability of their DNA-adducts.


Effects of prostaglandins and some anti-inflammatory drugs on the binding of 7,12-dimethylbenz[alpha]anthracene to the DNA of murine epidermal cells in culture.

The effects of prostaglandins (PG) E1, E2, F1 alpha and F2 alpha and some anti-inflammatory drugs such as acetylsalicyclic acid, flufenamic acid, indomethacin, and fluocinolone acetonide (FA) on the binding of [3H]7,12-demthylbenz[alpha]anthracene (DMBA) to DNA of murine epidermal cells have been investigated. PG E1 and E2 significantly inhibit the binding of DMBA to murine epidermal cells (MEC) DNA while PG F1 alpha and F2 alpha do not affec the binding. Salicyclic acid and flufenamic acid also do not alter the binding; whereas, indomethacin and FA lowered the binding of DMBA to DNA.


Benzo[alpha]pyrene antibody inhibition of benzo[alpha]pyrene-induced mutageneis.

An antibody to benzo[alpha]pyrene (BP) was prepared. The isolated antibody showed a specificity for BP and a low reactivity with another carcinogenic hydrocarbon, 7,12-dimethylbenz[alpha]anthracene (DMBA). The BP-antibody inhibited the in vitro cytotoxic and mutagenic activity of BP in both a rat embryo fibroblast- and a rat lung cell-mediated mutagenesis system. A possible correlation of these in vitro findings to the in vivo carcinogenesis situation is discussed.


Modification of the effect of a gonadoliberin analog on 7,12-dimethylbenz(a)anthracene-induced rat mammary tumors by hormone replacement.

A gonadoliberin analog, (D-leucyl6, desglycyl-NH2(10), prolyl ethylamide9) gonadoliberin, is known to suppress ovarian function and plasma prolactin levels. Its antitumor activity was evaluated against mammary tumors induced in Sprague-Dawley rats by dimethylbenz(a)anthracene. Observations were made when the analog, referred to as A-43818, was given alone and together with estrogen replacement or perphenazine, A-43818, 10 microgram s.c. twice a day for 6 weeks, was highly effective in producing tumor remissions. All of the 11 animals survived throughout the observation period, complete regressions occurred in 8 of 13 tumors, and 2 were classified as static. None of the 16 tumors  in 12 control rats regressed, and there were 4 deaths. When estradiol benzoate, 2 microgram s.c. each day, was administered with the A-43818, antitumor activity was suppressed; only 2 of 17 tumors regressed, 6 were static, and 5 of the 10 rats in this group died. Perphenazine, 1 mg i.m. daily, a dose known to cause hyperprolactinemia, also impaired the efficacy of A-43818. Three of 14 tumors regressed, 6 were static, and the rest continued to grow; 3 of the 12 rats died within 6 weeks of starting treatment.


Apparent rat strain-related sensitivity to phorbol promotion of mammary carcinogenesis.

It has been reported that twice-weekly i.p. injections of 4 mg phorbol for 10 weeks, after a single feeding of 6 mg dimethylbenz(a)anthracene (DMBA) in female Wistar rats, led to a significant augmentation of mammary adenocarcinoma incidence and of lymphatic leukemia incidence as compared to 6 mg DMBA alone. In an experiment reported here, in female Sprague-Dawley rats, using the same doses of DMBA and phorbol and the same injection schedule, phorbol given after DMBA did not augment mammary adenocarcinoma incidence or lymphatic leukemia incidence as compared to DMBA given alone. It thus appears that there is a strain-related sensitivity between Wistar and Sprague-Dawley rats with regard to the promoting activity of phorbol when phorbol treatment follows DMBA treatment, and mammary adenocarcinoma incidence and lymphatic leukemia incidence are studied. Further, in Sprague-Dawley rats, phorbol did not promote mammary fibroadenoma incidence in DMBA-treated rats, mammary adenocarcinoma incidence in procarbazine-treated rats, and mammary adenocarcinoma incidence or mammary fibroadenoma incidence in X-ray-treated rats. DMBA and procarbazine, with or without phorbol, tended to induce more mammary neoplasms in the anterior (thoracic) than in the posterior (abdominal) mammary glands. X-irradiation tended to induce mammary neoplasms in approximately equal numbers in the anterior and posterior mammary glands. It was suggested that regional differences in chemically induced mammary carcinogenesis were due to a difference in the transport and delivery of the chemical carcinogens to the regions rather than a difference in the amount of mammary gland tissue in the regions. An analysis of the numbers of Sprague-Dawley rats that developed either no mammary neoplasms, or only mammary adenocarcinomas, or only mammary fibroadenomas, or both mammary adenocarcinomas and mammary fibroadenomas in response to DMBA, procarbazine, and X-ray, suggested that the development of a mammary adenocarcinoma or the development of a mammary fibroadenoma are independent processes.


Inhibition of lymphocyte function in rats fed higher-fat diets.

Concanavalin A-induced blastogenesis of spleen lymphocytes was significantly inhibited when lymphocytes from rats on a high-polyunsaturated-fat diet were compared to lymphocytes from rats on a low-fat diet. Responsiveness was dependent on source of serum since lymphocytes from rats fed a low-fat diet were suppressed in serum from rats fed a high-polyunsaturated-fat diet. Alternatively, lymphocytes from rats on a high-polyunsaturated-fat diet were more responsive in serum from low-fat-fed rats compared to their response in autologous serum. One of the inhibiting factors in serum was the lipoprotein fraction; however, rats on a high-polyunsaturated-fat diet probably had additional inhibitors in their serum. While tumor incidence was highest in rats with the least responsive lymphocytes was highest in rats with the least responsive lymphocytes and lowest in rats with the most responsive lymphocytes, the significance of the observation is not known.


Serum tyrosinase in malignant disease, its activity, and the electrophoretic patterns of the enzyme as carried by immunoglobulins'.

Serum tyrosinase activity in many persons with metastatic diseases was found to be significantly higher than activity in normal persons. The highest activity was observed in melanoma and breast carcinoma. The electrophoretic patterns of serum tyrosinase, resolved by electrophoresis of a serum tyrosinase fraction followed by incubation of the gel sample with L-dopa, and represented as sets of RF's of melanin bands, were characteristically different in melanoma, breast carcinoma, and certain other diseases. The RF's of melanin and protein bands in the serum enzyme preparations from melanoma patients were concisely defined. Further, some potent serum fractions inhibiting tyrosinase melanogenic activity have been obtained, and the presence of tyrosinase inhibitors in the serum enzyme preparation has also been demonstrated. More detailed exploration of these serum tyrosinase parameters may provide more specific and sensitive detection for certain malignant diseases.


Phase II evaluation of hexamethylmelamine in advanced breast cancer: a Southwest Oncology Group study.

Ninety-eight patients with metastatic breast cancer, heavily pretreated with other agents, were entered in a phase II trial of hexamethylmelamine (HEX). Patients were randomized to receive HEX alone or combined with prophylactic pyridoxine. There was a 2% response rate in 89 partially or fully evaluable patients. Seven percent of these patients developed neurologic toxicity which occurred in the HEX-alone group only.


Hexamethylmelamine in ovarian cancer resistant to cyclophosphamide and adriamycin.

Thirty-four patients resistant to cyclophosphamide and Adriamycin received hexamethylmelamine at one of two dose regimens: 6 mg/kg/day orally for 21 days every 4 weeks or 8 mg/kg/day orally for 21 days every 6 weeks. Only five patients responded. Tolerance to the drug was, however, satisfactory; nausea, pyrosis, and vomiting were the only frequent side effects. Myelosuppression occurred in only one case.


Phase II trial of pyrazofurin, alone and in combination with trifluorothymidine, in non-Hodgkin's lymphoma.

A phase II trial of pyrazofurin, alone and in combination with trifluorothymidine, was carried out in patients with advanced non-Hodgkin's lymphoma. None of the 19 patients evaluable for response had complete or partial remissions but 14 had minor regressions. Toxicity, consisting primarily of myelosuppression and stomatitis, was of moderate severity. These studies suggest that pyrazofurin, alone or in combination with trifluorothymidine, is of limited utility in advanced non-Hodgkin's lymphoma.


The structure of the capsular polysaccharide from Cryptococcus neoformans serotype D.

The capsular polysaccharide from Cryptococcus neoformans serotype D has been studied by employing the usual methods for the elucidation of chemical structure. The results are consistent with the occurrence of a polysaccharide having both D-glucosyl-uronic acid and D-xylosyl groups present as nonreducing end-groups attached to O-2 of D-mannosyl residues which are linked alpha-D-(1 leads to 3) in a linear backbone.


Effects of nitroglycerin and dipyridamole on acute myocardial infarction: a thermographic study in the dog.

The effects on an acute myocardial infarction of equipotent hypotensive doses of nitroglycerin and dipyridamole were studied using the thermographic technique in open-chest dogs. Nitroglycerin did not reduce the cool ischaemic area in any of the nine dogs studied. In six of the dogs, in which the ischaemic region could be properly measured, the cool area slightly grew in size. Dipyridamole reduced the size of the ischaemic area in all the 20 dogs used. In the 19 in which the size could be adequately digitalised, the decrease in infarction size was statistically significant.


Immunofluorescence study of the preoptico-terminal LRH tract in the female squirrel monkey during the estrous cycle.

Reactive LRH neurons of the preoptico-terminal trace were studied with rabbit antisera to unconjugated or conjugated synthetic LRH in normally cyclic female squirrel monkeys. The specific immunoreactive material present along the preoptic-terminal LRH tract showed significant modifications that were particularly distinct (1) at the level of the perikarya, and (2) in the axons and axonal endings situated around the short and long capillary loops of the lamina terminalis. Conspicuous variations were observed during the luteal phase (with distinct and fast refilling), and the early and mid-follicular phases (suggesting a pulsatile mode of release); minimum concentrations were attained during the late and peri-ovulatory phases. A comparison is made between these modifications and those observed along the hypothalamo-infundibular LRH tract in the same animals. The preoptico-terminal LRH tract is thought to be one of the possible mechanisms controlling the sensitivity of gonadotropic cells to LRH, and the synthesis and storage of gonadotropins before their mid-cycle discharge.


Influence of thyroid hormone on the development of peoptic-hypothalamic monoaminergic neurons in tadpoles of Bufo bufo japonicus.

The effect of thyroid hormone on the development of diencephalic monoaminergic neurons was studied in tadpoles of Bufo bufo japonicus. Monoamine-containing neurons in the preoptic recess organ (PRO) appeared later than those in the paraventricular organ (PVO) and nucleus infundibularis dorsalis (NID). After deprivation of thyroidal primordium no fluorescent neurons developed in the PRO. Development of monoaminergic neurons in the PVO and NID was not affected by thyroidectomy. Thyroxine treatment brought about the fluorescent neurons in the PRO of the thyroidectomized tadpoles. Fluorescent terminals in the median eminence became conspicuous around the capillaries which penetrated to the median emience, when the tadpoles reached late prometamorphic stage. In the median eminence of thyroidectomized tadpoles, the monoaminergic axon terminals did not develop. Thyroxine induced both the fluorescent terminals and the capillary penetration in the median eminence of the thyroidectomized tadpoles. In the tadpoles hypophysectomized at tail-bud stage, thyroxine induced neither the fluorescent terminals nor the capillaries in the median eminence.


Post-replicative nonribosomal transcription units in D. melanogaster embryos.

We have analyzed transcription units from D. melanogaster embryos in which most cells were in either the late S or the G2 phase of a synchronous cell cycle. Much of the chromatin of these embryos was observed in the form of parallel strands which appear to be newly replicated sister chromatids (McKnight and Miller, 1977). Lateral ribonucleoprotein (RNP) fibers were commonly observed attached to chromatin strands and are believed to be nascent transcripts. Very similar lateral fiber arrays were found on sister chromatid strands. Such paired, post-replicative RNP fiber arrays offer a unique opportunity to compare the transcription patterns on two copies of the same transcription unit. Analyses of paired transcription units yield the following conclusions. Transcription initiation and termination sites are the same for both copies of each transcription unit. Each transcription unit has a characteristic and constant RNP fiber frequency, and the fiber frequency is identical for both copies of the transcription unit. Active transcription units can have internal, fiber-free gaps which may result from interruptions in initiation of transcription. Transcription units differing in polarity and fiber frequency can occur in immediate juxtaposition. The relevance of these findings to eucaryotic transcriptional control mechanisms are discussed.


Deletion mapping of two D. melanogaster loci that code for the 70,000 dalton heat-induced protein.

Using deficiencies in D. melanogaster that lack either the 87A or 87C heat-induced puffs, we have shown that the 70,000 dalton heat-induced protein (hsp 70) is encoded at both these loci. Embryos deleted for one of the two loci retain the ability to make hsp 70 after heat shock, but deleting both loci eliminates synthesis of hsp 70. Thus both loci encode hsp 70 and can be active following heat shock. We have analyzed the proteins made by embryos lacking either 87A or 87C, and have compared the 87A- and 87C-coded hsp 70 by isoelectric focusing and tryptic peptide fingerprinting. The hsp 70 made by the two loci is very similar, although a variant tryptic peptide appears to be encoded only at 87C. Using deficiencies with slightly different breakpoints, we have mapped the 87A locus to band 87A7, the site of the 87A heat-induced puff. The 87C locus maps within 87C1.


Differential replication of ribosomal gene repeats in polytene nuclei of Drosophila.

The genes coding for the 18S and 28S rRNAs in D. melanogaster were examined using Southern transfers of DNA from diploid or polytene tissue. A ribosomal gene repeat 12 kb in length is present in DNA from diploid tissue of males and is the major repeat on the Y chromosome. This repeat is present in low amounts on the X chromosome, which contains major repeats of 17 and 11.5 kb. In polytene nuclei of males, the 12 kb band is disproportionately replicated, and only a very low amount of the 11.5 kb repeat and no 17 kb repeat are detected. Polytene nuclei of females contain reduced amounts of the 17 kb repeat relative to the 11.5 kb repeat. This disproportionate replication of specific ribosomal gene repeats suggests that polytenization of the rDNA may involve an extrachromosomal mechanism. Evidence that genes from only one nucleolus organizer are replicated during polytenization in X/Y and X/X flies is discussed. A method for analyzing DNA from tissue of individual larvae was developed to test for population heterogeneity in ribosomal gene structure. Heterogeneity was observed in the ribosomal genes of three Ore R lines, four other D. melanogaster strains and between males and females of the same strain.


Evolution of transcribed and spacer sequences in the ribosomal RNA genes of Drosophila.

Examination of the ribosomal RNA (rRNA) gene of six sibling species that make up the D. melanogaster subgroup reveals that the nontranscribed spacer is highly conserved during evolution. Indeed, the spacer is at least as conserved as the transcribed rRNA sequence in four of the six species and only slightly less conserved in the others. These data support the hypothesis previously suggested (Tartof and Dawid, 1976) that selection has a significant role in maintaining the parallel evolution of genetically separate but homologous redundant gene clusters.


Satellite Ic: a possible link between the satellite DNAs of D. virilis and D. melanogaster.

In this study, we isolated and characterized a previously undetected cryptic satellite DNA comprising 0.1% of the total nuclear genome of D. virilis. This satellite is hidden from detection in neutral CsCl by satellite I and is therefore designated cryptic satellite I or Ic. Sequence analysis reveals that Ic is the repeating heptanucleotide [poly d(AATATAG): d(CTATATT)]. It is more closely related to the three simple sequence satellite DNAs of D. melanogaster, a distantly related species, than it is to any of the major D. virilis satellite DNA sequences. Ic may therefore be a link between the simple sequence satellites of D. virilis and D. melanogaster. As an extension of this theory, we have constructed a "family tree" linking the satellites of D. virilis and D. melanogaster by a series of "simple" operations. Only one intermediate required by this evolutionary scheme has not yet been identified.


Isolated liver perfusion--a tool in mutagenicity testing for the evaluation of carcinogens.

An isolated liver perfusion system suitable for the combination with Chinese hamster V79 cells is described. With this system, it is possible to study, with the V79 cells as genetic targets, the mutagenic effect of a chemical after metabolic activation in the intact organ. Those substances commonly used in mutagenicity testing as inducers of drug metabolising enzymes, i.e. Arochlor 1254. Phenobarbital(PB) and 3-Methylcholantrene(3-MC), were studied for their effect in the isolated perfused liver. PB increased the bile flow, which was not significantly affected by the other inducers. Only Arochlor caused a significant increase in the amino acid incorporation into plasma proteins and total liver proteins (expressed per mg liver protein). None of the inducers had an effect on gluconeogenesis from lactate or urea synthesis. All three inducers caused an increase in the level of microsomal P-450 enzymes, the biggest increase being seen after Arochlor-induction (170%), followed by PB(90%) and 3-MC(50%). Arochlor- and PB-induction had a dramatic effect on N- and C-oxygenation of N, N-dimethylaniline: N-oxygenation was decreased by 35% and 40% respectively and C-oxygenation increased by 130% and 140% respectively. The advantages of the isolated perfused liver as an intact metabolising unit is discussed in relation to other mutagenicity assays, in which subcellular fractions are used as the metabolising system.


Fluorescence of hydrocarbon-deoxyribonucleoside adducts.

In comparison with the fluorescence emission spectra of 7-methylbenz[a]-anthracene-nucleoside adducts, the fluorescence emission spectra of hydrocarbon-deoxyribonucleoside adducts containing a methyl substituent in the "bay region" lack spectral resolution at room temperature and appear at substantially longer wavelength. This spectral resolution is improved when spectra are measured at 77 K and an irreversible spectral shift to shorter wavelength, accompanied by improved resolution, results from mild acid hydrolysis. These spectral properties peculiar to the "bay region-substituted" adducts presumably result from an intramolecular interaction between the hydrocarbon fluorophore and the attached nucleoside brought about, in the examples studied here, by the presence of the 12-methyl group in 7,12-dimethylbenz[awanthracene (DMBA) and in 7-hydroxymethyl-12-methylbenz[a]anthracene. This interaction suggests that the site of nucleoside attachment is in close proximity to the 12-methyl group and that binding occurs, therefore, through the intermediacy of a 3,4-diol-1,2-oxide, i.e. a "bay region" diol-epoxide in each case.


In vivo binding of 2,3,6,2',3',6'-hexachlorobiphenyl and 2,4,5,2',4',5'-hexachlorobiphenyl to mouse liver macromolecules.

The role of metabolic activation in the binding of polychlorinated biphenyls (PCBs) to cellular macromolecules was investigated in vivo by comparing the relative binding of 2,4,5,2',4',5'-[U-14C]hexachlorobiphenyl (2,4,5), a slowly metabolized PCB, with that of 2,3,6,2',3',6'[U-14C]hexachlorobiphenyl (2,3,6), a rapidly metabolized PCB, and the appropriate controls. Each hexachlorobiphenyl was administered to mice, orally for 5 days (7.28 mg/kg/day). Following the dosing schedule, animals were killed at 1, 5 and 8 days. The concentration of each PCB was determined in liver, muscle and kidney and in purified macromolecules isolated from those tissues. The concentration of 2,4,5 was consistently higher than the concentration of 2,3,6 in all tissues studied. However, the amount of 2,3,6 bound to the purified macromolecules was consistently at least one order of magnitude greater than that of 2,4,5. The greatest binding was observed in RNA followed by protein and DNA, respectively. The purity of the macromolecules and the presence of PCB-derived radioactivity at the monomer level were confirmed. This is the first report of 14C-labeled PCB being bound to purified RNA, DNA, and proteins isolated from the tissues of animals treated in vivo. The binding is thought to be covalent and to be the result of metabolic activation.


[Existence of different populations of unitary evoked postsynaptic potentials at the frog neuromuscular junction].

The postsynaptic response to monoquantic evoked transmitter release (mean quantal content about 0.3) has been studied at temperatures from 10 to 23 degrees C. The delay between nerve stimulation (1 to 10 Hz) and the unitary postsynaptic potentials fluctuates by steps. The existence of preferential delay sites can always be detected (mean number 13.5 +/- 3.1). Identical delay unitary postsynaptic potentials often shows identical amplitude and identical time to peak. These results suggest that few emitting sites are preferentially activated along the nerve terminal at low level release during long lasting stimulation. The "single process" assumption used in statistical studies of transmitter release is probably oversimplified.


[Inhibitory action of estradiol on testosterone production in rat embryo].

Embryos from Rats given an injection of 0,5-1 mg of oestradiol benzoate on days 17--19 of pregnancy show a 3--4-fold reduction in their testicular testosterone content with respect to the control embryos, 24--48 hrs later. OEstradiol most likely acts by inhibiting LH secretion, which in turn results in a lowered testosterone production.


[Association of brain glycoprotein (NSA 3) with neuronal membranes and with the nodes of Ranvier].

The localisation of brain glycoprotein NSA 3 was studied by means of indirect immunofluorescence on alcohol fixed, paraffin embedded sections of Rat brain. These techniques allowed the localisation of NSA 3 to the membrane of some (about 10%) of the neurons. In the white matter, the patterns were in agreement with the localisation of the Ranvier nodes. The nodes of Ranvier were also stained in peripheral nerves.


[In vitro inhibition of tRNA methyltransferases by queen substance, a pheromone of queen honeybees].

The Queen Substance 1, a pheromone of the queen Honeybee Apis mellifica is an in vitro inhibitor of E. coli B tRNA methylations. This activity is not specific of the methylase source, as inhibitions have been observed with preparations from queen honeybee ovaries, Rat liver or a Mouse plasmocytoma 1-adenine methylase. These results, together with preceding ones concerning t, t-farnesyl-acetone 3, are discussed.


[Consumption of vitamin A in intact and castrated rats. Relation to tocopherol and cytochrome P 450].

There are significant differences between castrated and non castrated male Rats in regard to vitamine A hepatic storage and plasma level. Cytochrom P 450 levels are the same in both groups. Those results are discussed.


[Identification of cytotoxic cells responsible for lysis of target cells pretreated with concanavalin A in spleen populations of pregnant mice with suppressive activity].

During an alloimmunization, killer cells which lyse target cells only in the presence of a lectin are generated. That these cells, as well as suppressive cells, share immunocytological properties with specific killer cells, leads to the hypothesis that these cells may be concerned with the mechanism of immunosuppression. Two experimental results presented in this paper are consistent with this hypothesis: 1) Spleens from H-2k mice pregnant by H-2d males which bear a high suppressive activity also contain a relatively large number of killer cells having the ability to lyse Concanavalin A treated target cells and 2) supernatants of suppressive systems generated through an MLC block the cytolysis of specific target cells by the bound killer cells.


[Are rat alveolar macrophages capable of phagocytizing pollen grains?].

Plane-tree pollen grains were incubated in vitro with alveolar macrophages from inbred Rats, and the possibility of phagocytosis was investigated. No phagocytosis was observed even after 48 hrs incubation. But alveolar cells were bound to pollen grains generally at the apertures. This binding did not require the Fc receptor on the macrophage membrane.


[Topography of soluble gliofibrillar protein (GFA) in aged human brain].

A great variation was observed in the amount of soluble GFA depending on the brain area. Large amounts were found in the chiasma and in the pons whereas the cortical grey matter was poor. It is suggested that soluble GFA represents a potentiality of defense of the nervous tissue.


[Fragmentation of myosin A-1 light chain of fast muscle by trypsin].

Limited proteolysis of myosin by such proteolytic enzymes as trypsin, chymotrypsin or papain produces typical fragmentation of its heavy chain. Presently evidence is given that trypsin treatment cleaves the alkali light chain A-1 (20,700 dalton) to a shorter (ca 20,000 dalton) chain. The two "essential" thiols (SH-1 and 2) of moysin were alkylated with 17-C-N-ethylmaleimide and a non-negligible amount of radioactivity was also found in the two alkali light chains. Using the specific radioactivity of alkali light chain A-1 it was possible to identify it among heavy chain fragmentation products. The molecular weight of the newly formed A-1 indicates that limited tryptic cleavage of this A-1 confers on it a closer similarity with alkali light chain A-2.


[Activity of rabbit heart on thromboxane synthetase].

The soluble and microsomal fractions of Rabbit myocardium are not able to induce the synthesis of thromboxanes. On the contrary, they inhibit the thromboxane synthetase of various sources. The chemical structure of the active constituent responsible for this activity is not yet known: it is probably neither of an enzymatic nature, nor a protein of high molecular weight.


[Elaboration of an in vivo model, in half life size, for aeronautical purposes].

Toxic effects of combustion products originating from plane cabin constituents are studied, in vivo, in Rats. The model associates an important combustion room, and an experience room containing eight revolving cages for Rats; these cages allow a remote control of the movements of the Rats which can be electrically stimulated and ejected. Such a device permits to study the incapacitance, i.e. the inability to move. For the combustion of a seat, without ventilation, incapacitation appears in 2 to 6 min. Biochemical screening demonstrates on increase of pCO2, carboxyhemoglobin and glycemia and a decrease of pO2.


[K-Lymphocyte cytotoxicity: role of human blood group ABO, rhesus and P alloantibodies].

Using human erythrocytes of known antigenic density, sensitized by ABO and Rhesus (D) alloantibodies, it is shown that K cell cytotoxicity mediated by peripheral blood lymphocytes is directly correlated with the number of IgG molecules specifically bound to the target cell surface. The lytic sensitivity of P1k but not P2 or p erythrocytes coated with anti-Tja (anti-P + P1 + Pk) antibodies, demonstrates that effector K cells are mainly triggered through the IgG ANTI-Pk component of such sera.


[Macroscopic and microscopic study of human corpse skin grafting on athymic mice nu/nu].

32 thin grafts (500 to 700 mu) of different human corpse skin set apart during the first 24 hrs after the death have retaken on athymic Mousse whatever the reason of death and the age of the donor. 20 thick grafts have retaken at 30%. These skins preserve the aspects and behaviour of normal human skin but after 1 month they have a heavy melanic hyperpigmentation. These skins preserve their normal histological aspect during the first 3 days, then, when revascularisation is setting in, superficial areas of epidermic mortification, opposite dermal hypovascularised zones, appear. Between the 15th and the 21st day the epidermic alterations disappear and a heavy melanic hyperpigmentation is set up. The revascularisation is certainly the result of the human dermal vessels repermeabilisation. Till the 76th day neither dystrophy, dysplasia nor important inflammatory process exist.


[Comparative study, in the Allium test, of the cytotoxicity of tubulosine and of 6 synthetized pseudo-tubulosines].

Comparing cytotoxicity of tubulosine and of six newly synthesized pseudo-tubulosines in the Allium test, we have characterized all the current features of protein inhibition. Nevertheless cytotoxicity of the six pseudotubulosines is much weaker. Furthermore, "floating poles anaphasis", a disturbance characteristic of the action of tubulosine, has been found with a lesser intensity with the other compounds.


[Synthesis of juvenile hormones in vitro by the corpora allata of 5th stage larva of Locusta migratoria migratorioides (R and F) (Insecta, Orthopteroida)].

Corpora allata of Locusta migratoria 5th stage larvae synthesize J.H.1, J.H.2 and J.H.3 in vitro. The C.A. of insects of different ages exbit different rates of J.H. synthesis. J.H.1 and J.H.2 synthesis is less than 1 ng/48 h/gland. During the same time the J.H.3 production may be as much as 25.6 ng/gland. J.H. synthetic activity is the same between right and left C.A. The release of J.H. from the C.A. occurs immediately following synthesis. These results are compared with in vivo haemolymphatic J.H. levels.


Hydrolase activities in the rat aorta. III. Effects of regular swimming activity and its cessation.

It is possible that one of the consequences of regular physical activity could be a change of vascular metabolism. We studied the effects of regular swimming activity on specific activities of aortic hydrolases of male rats. Enzymes included: neutral alpha-glucosidase and lysosomal beta-galactosidase, N-acetyl-beta-glucosaminidase, cathepsin C, acid alpha-glucosidase, and acid cholesteryl esterase. After 8 or 16 weeks of a 1-hour/day swimming protocol, specific activities of four of the six aortic enzymes studied were increased over control levels, increases ranging from 7 to more than 42%. Acid cholesteryl esterase was one of the enzymes most affected by the exercise, increasing 25-30% above control levels. An 8-week sedentary period, after 8 weeks of a swimming regimen, resulted in return of the activity of acid cholesteryl esterase, but not those of the other hydrolases, to control levels. Decreases in body weight, blood pressure, and serum lipid levels also occurred in the swimming rats. Weight reduction per se was excluded as an explanation for the increases in aortic enzymes or decrease in serum cholesterol found with swimming. These findings show that regular physical activity is yet another factor with discrete and significant effects on the catabolic activity of vascular tissue.


Epicardial mapping and electrocardiographic models of myocardial ischemic injury.

The amplitude and distribution of epicardial ST-segment elevation (ST) were examined for an 8-hour period after coronary occlusion in eight baboons and five pigs. ST was determined from unipolar epicardial electrograms obtained from a high-resolution matrix of fixed electrodes overlying a transmural region of ischemia. A relatively uniform degree of ST was observed overlying the ischemic region for 20 minutes after coronary occlusion. A gradient in ST from the periphery to the center of the ischemic region was documented after 20 minutes of ischemia. In 10 other pigs, change in the degree of ST was examined contingent on either an increase (five pigs) or decrease (five pigs) in the size of the ischemic region after 1 hour of preexisting ischemia. An abrupt increase in the number of electrodes that showed ST (NST) from 7.8 +/- 1.24 (SEM) to 14.8 +/- 1.35 (90%) was associated with an increase in mean ST of 58% from 4.28 +/- 0.61 mV to 6.78 +/- 0.84 (p less than 0.05). An abrupt decrease in NST from 25.2 +/- 2.63 to 14.6 +/- 2.22 (42%) was associated with a decrease in mean ST of 24%, from 8.2 +/- 0.36 mV to 6.3 +/- 0.30 mV (p less than 0.01). The results during early ischemia (less than 20 minutes of ischemia) are accurately represented by a model of ischemia in which injury current arises only at the ischemic boundary. The results during later ischemia (after 20 minutes of ischemia) may be represented by a model in which ST is considered dependent on injury currents generated throughout the ischemic region.


Enzyme linked immunosorbent assay (ELISA) for IgG and IgE antibodies to protein and polysaccharide antigens of Aspergillus fumigatus.

ELISA has emerged as a useful alternative to other more costly and complex tests. Polystyrene microhaemagglutination plates have been used as solid phase to absorb Aspergillus fumigatus protein and polysaccharide components for detection of specific antibodies in patients with various forms of pulmonary aspergillosis. IgG and IgE antibodies to the polysaccharide as well as the protein allergens have been found. For the IgE test a double antibody technique has been developed, which is more sensitive than the conventional indirect ELISA.


The evaluation of selected parameters of immune function in asthmatics after long-term corticosteroid therapy.

The availability of inhaled beclomethasone diproprionate permitted the discontinuation of continuous, long-term systemic corticosteroid therapy (SCT) in a group of asthmatics who had previously required SCT to control their asthma. Twelve patients had been on SCT for a period of 2-22 years with an average duration of 7 years. To determine whether this previous long-term SCT and the current use of inhaled beclomethasone diproprionate had an effect on leucocyte functions, a variety of studies reflecting T lymphocyte, B lymphocyte and granulocyte function was done. The results were compared with those of twelve asthmatic patients of similar age ranges who had never received steroids. Results showed that the two patient populations could not be differentiated on the basis of phytohaemagglutinin stimulation of lymphocytes, sheep erythrocyte rosette formation, IgG, IgA, IgM and IgE concentrations or granulocyte bactericidal activity. Delayed skin reactivity was present in both groups, with more positive reactions in the non-SCT group. Polymorphonuclear adherence values were slightly lower in the SCT female population using beclomethasone diproprionate. The latter two minor differences may be due to the previous SCT, the use of beclomethasone diproprionate or the limited population of patients studied. We conclude from these studies that the long-term use of SCT at the low doses required for control of asthma resulted in little permanent effect on the variety of lymphocyte and granulocyte functions tested.


Ribonucleic acid in plasma from normal adults and multiple myeloma patients.

Reports of the presence of RNA in human plasma have been numerous, often suggesting that RNA in plasma is correlated with human disease. We critically examined the methods for determination of RNA in plasma. Lack of method specificity has caused previous workers to overestimate plasma RNA concentrations by more than 50-fold. To isolate RNA from plasma, we used both a phenol-chloroform extraction and a modified Schmidt-Thannhauser procedure. We show that RNA in plasma can be identified and quantified by alkaline hydrolysis of the plasma extract and subsequent separation of the resulting 2'- and 3'-mononucleotides by "high-performance" liquid chromatography. We could not detect RNA in plasma from either apparently healthy, normal adults or multiple myeloma patients, but found 1.1 mg/L in the plasma of a patient with Waldenström's macroglobulinemia. Our method is useful for the specific determination of RNA in plasma and will detect as little as 600 micrograms/L.


Enzymic determination of fructose in seminal plasma by initial rate analysis.

We describe a spectrophotometric assay for fructose in seminal plasma. The method is based on reduction of fructose by a commercially available preparation of sorbitol dehydrogenase (EC 1.1.1.14), with the concomitant oxidation of NADH. The initial rate of NADH oxidation, which is proportional to the fructose content of seminal plasma, can be measured either with a recording spectrophotometer or by conventional two-point kinetic assay. The method was as accurate, precise, and sensitive as, and more specific and rapid than, currently used colorimetric (resorcinol) methods for fructose determination. Values (mmol/L) for fructose in seminal plasma from several species are: man, 9 +/- 2 (SD); cynamolgus monkey (Macaca fasicicularis)., 108 +/- 19; bull, 30 +/- 1; and rabbit, 13 +/- 4. These values agree with previously published results. We believe the method is appropriate for both research and clinical use.


Neuraminidase activity in liver and brain from patients with I-cell disease.

The activity of neuraminidase in liver and brain from I-cell disease (Mucolipidosis II) was investigated. Neuraminidase activities using two substrates [alpha-L-N-acetylneuraminosyl(2 leads to 3)lactose and alpha-L-N-acetylneuraminosyl(2 leads to 6)lactose] were reduced in the supernatant and sedimentable fractions obtained in isotonic KCl. The activity of beta-D-galactosidase was also reduced in the liver; on the other hand, both neuraminidase fractions were normal, although beta-galactosidase activities were markedly reduced. In view of these results, it is suggested that the defect of neuraminidase is not directly responsible for the primary etiology of I-cell disease.


"Lysosomal" enzyme activities in red blood cells of normal individuals and patients with homozygous beta-thalassaemia.

Four hydrolases, beta-galactosidase, beta-glucuronidase, beta-N-acetylglucosaminidase and acid phosphatase were examined in red blood cells (RBC) of normal donors and patients with homozygous beta-thalassaemia. Highly sensitive fluorimetric substrates were used to determine the specific activities of these enzymes. In order to avoid contamination by lysosomal activities derived from white blood cells (WBC), the mature RBV were separated from other blood elements by cellulose chromatography. The hydrolase activities in normal RBC were detected only in their plasma membranes and were found to be considerably lower than in WBC or platelets. In thalassaemic RBC, hydrolase activities were present in both plasma membranes and in the soluble fraction. The normoblast fraction contributed most of the hydrolase activity found in these preparations, suggesting the presence of lysosomal particles in thalassaemic RBC. No differences in the enzymatic activities were found when purified membranes of mature RBC from thalassemic and normal preparations were compared. The origin and roles of these hydrolytic enzymes in normal and thalassaemic RBC membranes are not known.


"Grand mal type" of spike discharge as interseizure phenomenon.

Grand mal type of sequential spike discharges at 8-12 cps are rare as interseizure phenomena. When the grand mal component is prominent, it usually indicates a greater tendency for major convulsions. The case of a 15 year old girl who developed akinetic seizures along with her menarche and whose electroencephalogram showed recurring sharp waves at 10-11 cps, is reported, and its clinical significance discussed.


Augmentation of pituitary thyrotrophin response to thyrotrophin releasing hormone during subphysiological tri-iodothyroinine therapy in hypothyroidism.

Five hypothyroid patients are reported with increased pituitary TSH response to TRH during administration of T3. In one patient treated with intravenous T3, 50 micrograms daily for 10 days, the peak serum TSH and total pituitary TSH reserve after TRH increased coincident with increases in serum T3 and T4 levels and a decrease in the basal TSH concentration. In four patients treated with oral T3, the peak serum TSH and total pituitary TSH reserve after TRH increased during administration of subphysiological doses of T3. Peak serum T3 levels occurred 4 h after ingestion and increased progressively with increasing T3 doses. Serum TSH levels decreased modestly with the nadir at 4 h after T3 ingestion and then returned to basal levels at 24 h. Augmentation of TSH responses to TRH occurred simultaneously with decreases in serum cholesterol, as well as increases in the pituitary prolactin response to TRH, and increase in the GH and cortisol response to insulin induced hypoglycaemia where these responses could be studied. These data demonstrated a positive effect of subphysiological T3 therapy in these hypothyroid patients on the TSH response to TRH as well as increases in the responses of other pituitary hormones to stimulation.


Pituitary and peripheral resistance to thyroid hormone.

A 32-year-old Caucasian male, clinically euthyroid, with paranoid schizophrenia and granulocytopenia, had elevated total and free serum T4 and T3; serum TSH was normal (2.7 +/- 0.7 micronU/ml). There was no goitre present, no evidence of Graves' disease, and no evidence of pituitary tumour. He had a normal response to methyl-TRH, with a TSH increment of 14.6 micronU/ml, T3 increment of 212 ng/dl, and T4 increment of 4.7 microgram/dl; baseline value and decreased the TSH increment in response to methyl-TRH. T3 therapy (100 microgram/day) decreased the thyroidal radioactive iodine uptake to less than half the baseline prolactin was normal with a normal response to methyl-TRH to 4.1 micronU/ml. Iodine therapy caused an increase in his baseline TSH with an increase in the TSH response to TSH. The metabolic clearance rates (MCR) and production rates (PR) of T3 and T4 were increased. Baseline serum levels of glycoprotein hormone alpha-subunit were normal and showed a slight increase in response to methyl-TRH, similar to normal subjects. This patient has evidence of partial pituitary and peripheral resistance to thyroid hormone; his only evidence for hyperthyroidism is the elevated MCR and PR of T3 and T4.


Longitudinal study or serum thyroid hormones, chorionic gonadotrophin and thyrotrophin during and after normal pregnancy.

Measurements of serum levels of thyroxine (T4), free T4, 3,5,3'-triiodothyronine (T3), free T3, 3,3',5'-triiodothyronine (reverse T3, rT3), thyroxine-binding globulin capacity (TBGcap), chorionic gonadotrophin (hCG) and thyrotrophin (TSH) were carried out prospectively in eight women with uncomplicated pregnancies, in order to examine interrelationships between the thyroid gland and thyroid stimulating hormones during pregnancy. During pregnancy the levels of T4, free T4, T3, rT3 and TBGcap were significantly elevated, and TSH was decreased. It was noted that the elevation of T4 was maintained from the 8th to the 27th week of gestation while the level of TBGcap progressively increased. The levels of free T4 and rT3 in the first and third trimesters were significantly higher than those of age-matched, non-pregnant women. The levels of hCG showed a biphasic variation, with a peak in the 8th to 15th weeks, followed by a decline in the second trimester and a small, secondary elevation in the 32nd to 39th weeks. This later elevation was positively correlated with changes in free T4 and free T3 levels. The increase of serum T4 accompanied by an increase of free T4 in the first trimester appeared due to augmented secretion of T4, rather than being secondary to the elevated levels of TBGcap.


Progressive visual failure in acromegaly following external pituitary irradiation.

Four out of twenty-three acromegalic patients selected for treatment with external megavoltage pituitary irradiation between 1961 and 1975 developed progressive visual failure. They had received megavoltage external irradiation through multiple portals from a cobalt-60 unit over a period of 3 weeks. Visual deterioration began 2 months to 6 years after irradiation. In two patients the optic nerves were explored. In both, post-mortem later confirmed radiation damage to the optic nerves and hypothalamus. In one case there was also necrosis of the right frontal lobe with necrosis and inflammation of the bone surrounding the pituitary fossa. In the two other patients, extensive clinical and neuroradiological investigation excluded the presence of a suprasellar mass as a cause for this visual failure and a clinical diagnosis of radiation necrosis was made.


Bromocriptine suppression of TRH-stimulated prolactin and thyrotrophin release and accompanying inhibition of bromocriptine induced growth hormone release by TRH in normal man.

Six normal fasting males received on four separate occasions in random order (1) a placebo tablet followed 60 min later by 200 microgram of TRH intravenously (2) bromocriptine 2.5 mg orally followed by TRH intravenously (3) bromocriptine 2.5 mg orally followed by a placebo injection and (4) placebo tablet followed by placebo injection. Plasma prolactin and TSH responses to TRH were decreased following bromocriptine pretreatment. The rise of plasma growth hormone after bromocriptine was inhibited by TRH. The rise in plasma FSH seen after TRH injection was not influenced by bromocriptine pretreatment. Circulating LH and insulin concentrations were unaffected by any drug administration. These results suggest a dopaminergic influence on prolactin and TSH release in normal men, an inhibitory effect of TRH on bromocriptine stimulated growth hormone secretion, and no dopaminergic modulation of basal insulin secretion.


Maternal serum prolactin and its response to TRH in normal and complicated early pregnancy.

Maternal serum prolactin levels (PRL) were measured by radioimmunoassay in thirty-four women with either normal or complicated early pregnancy. The basal PRL level (mean +/- S.D.) of 33.4 +/- 16.4 ng/ml in normal pregnancy (n = 15) was similar to the level of 32.7 +/- 18.8 ng/ml in threatened abortion (n = 11) and 32.8 +/- 16.9 ng/ml in hyperemesis gravidarum (n = 8). Two patients, one with blighted ovum and the other with subsequent spontaneous abortion, demonstrated PRL levels lower than the range of 20-63 ng/ml in the control group. The PRL response to 200 microgram of synthetic thyrotropin releasing hormone (TRH) administered intravenously was similar throughout the patient groups. The basal level of PRL in the whole series was more closely related to the level of serum oestradiol (r = 0.778, P less than 0.001) than to that of serum progesterone (r = 0.442, P less than 0.05). However the increments of PRL following TRH administration did not correlate with either oestradiol or progesterone.


Effects of heroin addiction on thyrotrophin, thyroid hormones and porlactin secretion in men.

Pituitary-thyroid function in male heroin addicts and addicts after abstinence (ex-addicts) was studied and compared with that of healthy euthyroid men. In heroin addicts the increases in circulating total thyroxine and triiodothyronine levels were accompanied by an increase in the thyroid hormone uptake test. These changes may reflect a quantitative increase in thyroxine binding globulin. Reverse triiodothyronine concentrations in heroin addicts were normal. The thyrotrophin-releasing hormone elicited a diminished thyrotrophin response in heroin addicts which was significantly different from that in control subjects and ex-addicts. An elevation of serum prolactin was noted in heroin addicts, while ex-addicts had normal levels. Gradual recovery of pituitary-thyroid function occurred after heroin withdrawal.


Pituitary microadenoma: diagnostic studies.

Forty-one women with oligo-menorrhoea and/or galactorrhoea were subjected to hypothalamic pituitary-thyroid testing in an attempt to establish the presence or absence of an underlying pituitary microadenoma. They were divided into two groups in accordance with the serum level of prolactin (PRL): Group I (N = 25, mean +/- SE 17.6 +/- 1.5 ng/ml) and Group II (N = 16, 102.8 +/- 29.7 ng/ml). The dynamic tests performed were a TRH test, a stimulation test with metoclopramide (MCP) and a suppression test with bromocriptine. The results of these tests were compared with those obtained in nine normal women and eleven patients with surgically proved pituitary microadenoma. Radiologically abnormal pituitary fossas were found in ten subjects from Group I and in fourteen from Group II. All patients were euthyroid. A persistently elevated serum TSH in response to TRH was observed in patients of Group II suggesting an hypothalamic abnormality and a progressive decrease in the 120-min use of serum T3 was noted with increasing evidence of the existence of a pituitary tumour. A negative correlation was found between the basal serum PRL and the rise of serum PRL with TRH. Patients from Group II showed a lower PRL response to MCP when compared to Group I and again a negative correlation between basal level of serum PRL and the change after MCP was observed. No clear difference in the 4-h response to bromocriptine was found between the different groups of subjects. In conclusion, none of the three tests analysed permitted us to establish which of the patients had an underlying pituitary microadenoma.


Local invasion by laryngeal carcinoma--the importance of focal (metaplastic) ossification within laryngeal cartilage.

Patterns of cartilage invasion by squamous carcinoma were examined in 34 consecutive laryngectomy specimens with particular reference to selective involvement of ossified cartilage. Direct infiltration of the laryngeal framework was demonstrated in 17 cases--16 (out of 17) transglottic carcinomas and in a simgle example of a combined glottic and infraglottic tumour. The susceptibility of ossified laryngeal cartilage to tumour invasion was confirmed, and morphological studies ahve clarified the underlying mechanisms. Invasion is a largely indirect process dominated by local bone destruction by osteoclasts, operating in front of the advancing tumour. One established, carcinoma cells infiltrate and erode bone alone, and the osteoclasts disappear. Reasons for the particular susceptibility of ossified laryngeal cartilage to tumour invasion are discussed and attention is drawn to the role of the tumour-associated osteoclast activating factors such as postaglandins. Therapeutic implications of cartilage invasion are noted.


Long-term complications in Hunter's syndrome.

This report describes the clinical and investigative findings in a 31-year-old man with Hunter's syndrome, showing evidence of severe multisystem involvement. Papilloedema has been present for at least 8 years with no evidence of progressive visual impairment. His most serious and life-threatening complications are laryngeal oedema and tracheal narrowing, which both compromise his respiratory function and constitute a major anaesthetic risk.


A suggestion of linkage between the Marfan syndrome and the rhesus blood group.

The linkage relationships of the locus for the Marfan Syndrome were investigated with 19 marker loci in 17 families. Close linkage with several marker loci was excluded. There was a suggestion of linkage between the locus for the Marfan Syndrome and Rh.


Immunodiagnosis of alpha chain disease.

Since the early diagnosis of alpha chain disease (alphaCD)) is essential to successful treatment and to epidemiological studies, the available immunodiagnostic techniques were compared for their sensitivity, specificity and ease of performance on a panel of sixteen sera, comprising ten alphaCD sera and six control sera containing either IgA myeloma protein or high levels of polyclonal IgA. Immunoselection by immunoelectrophoresis into gel containing a specially developed anti-Fabalpha antiserum provided the most sensitive and specific detection system for alphaCD protein. The same technique using anti-light chain antiserum for immunoselection was also highly sensitive, but proved less specific, being prone to false positives with difficult IgA myeloma proteins. Somewhat less sensitive, but specific and simple to perform, was immunoelectrophoresis using an antiserum recognizing the conformational specificities of Fabalpha as well as those of the constant region of alpha chains. Immunoselection using the Ouchterlony or rocket techniques proved to be less sensitive and prone to false positives when some IgA myeloma sera were tested.


Effect of gold sodium thiomalate on murine lymphocyte functions.

The in vitro effects of gold sodium thiomalate (GTM) on various murine splenic lymphocytic functions were tested. The presence of GTM in cultures of splenic cells suppressed anti-hapten responses to both thymus-independent and thymus-dependent antigens. GTM also suppressed the in vitro generation of cytotoxic effector cells as well as the mitogenic response to both T cell and B cell mitogens. This suppression could not be reversed by the addition of irradiated spleen cells. Spleen cells exposed to GTM for 4 hr prior to culture also exhibited similarly suppressed functions, although their functional capacity could be fully restored by the addition of irradiated spleen cells. These results show that GTM inhibits both humoral and cellular immune mechanisms and appears to act primarily at the accessory (macrophage) cell level, with perhaps a secondary effect on T lymphocytes.


Ski boot compression syndrome.

The extensor tendons and peroneal nerve can be compressed at the ankle by the tongue of the ski boot. The resulting neuritis and synovitis may be severe enough to mimick an anterior compartment syndrome. Treatment consists of conservative methods but the paresthesiae may remain for long periods of time.


Treatment of mucopolysaccharidosis: clinical and biochemical aspects of leucocyte transfusion as compared with plasma infusion in patients with Hurler's and Scheie's syndromes.

The therapeutic effectiveness of leucocyte transfusion (LT) was compared with that of plasma infusion (PI) clinically by range of motion (ROM) of joints and biochemically from the standpoint of alpha-L-iduronidase activity and urinary excretion of acid mucopolysaccharides (AMPS) in 2 patients with Hurler's and Scheie's syndromes. Both syndromes are considered to be due to the lack of alpha-L-iduronidase activity, a congenital metabolic disorder. As a result, leukocyte transfusion surpasses plasma infusion with respect to enzyme content, the grade and duration of clinical improvement in the stiffness of joints. Clinical improvement in the stiffness of joints was correlated with the degradation of AMPS when the ratio of urinary AMPS fragments to the total large molecule AMPS has become 50% or more after the leucocyte transfusion and plasma infusion.


"Normal" talar tilt angle.

A study to determine the degree of normal talar tilt in individuals with no history of ankle injuries was performed on 404 ankles of 202 midshipmen at the U.S. Naval Academy. Manual inversion stress roentgenograms were used to test this group of young healthy adults, most of whom were males. Of the 404 ankles tested, only 39 (9.7%) had any measurable talar tilt and only 7 ankles (1.7%) had a tilt over 5 degrees. Normal talar tilt does not range to 23 degrees. A normal ankle in a healthy young adult with no history of trauma has a small probability of having any talar tilt. When stress roentgenograms are used to evaluate a fresh lateral ankle ligament sprain in this type of individual, there is a very high probability that talar tilt over 5 degrees would represent significant injury to one or more of the lateral supporting structures.


Ossification of the posterior longitudianl ligament of the cervical spine: subtotal vertebrectomy as a treatment.

If the ossification is localized in the central part of the intervertebral space, anterior decompression surgery of Cloward may be indicated. However, if most posterior ligamentous ossification covers the posterior surface of the vertebral body and the lateral sides, Cloward's method is not indicated. We have developed a method of anterior decompression and fusion surgery to clear the ossified area, resecting the vertebral body (subtotal vertebrectomy). The results were excellent in 3 of 4 patients with preoperative radiculopathy; there were 2 excellent and 16 good results in 22 patients with myelopathy. Of course laminectomy can produce some good effects, but the anterior obstruction removes through the anterior approach. Anterior decompression may be indicated when ossification is localized below the C3--4 intervertebral space, and when the spinal canal is not narrowed in the upper cervical region, even if ossification extends over the whole cervical spine. Laminectomy is advised when ossification involves all levels of the cervical spine, the upper cervical spine is narrowed or the sagittal dimension of the spinal canal is reduced more than 60%.


Formation of a bone and joint following blunt injury to the pelvis.

The formation of a new bone from the lateral wall of the pelvis following blunt injury occurred in a 55-year-old woman. The bone articulated with the top of the trochanter by means of a synovial-lined joint. The case is an example of the remarkable ability of external forces to remodel newly formed bone into a functional architectural unit and to produce an accessory pseudojoint.


Drugs and pollutants in breast milk.

Mechanisms of transport of environmental pollutants and ingested drugs in breast milk and their absorption in newborn nurslings are outlined. The authors urgently call for additional research to determine which maternal medications are safe for nursing infants.


Distribution of endogenous cobalamin between the transcobalamins in various mammals.

1. Plasma samples from ten mammals were chromatographed on Sephadex G-200 columns and the total cobalamin content of each fraction was determined. 2. Unlike the situation in man, the bulk of the endogenous plasma cobalamin was found attached to a transcobalamin II-like protein in all ten animals. Transcobalamin 0 carried between 3 and 20% and this proportion appeared to be inversely related to the plasma total cobalamin. No endogenous cobalamin peak corresponding to transcobalamin I was detected in any species, though in the rabbit 5.3% of the plasma total cobalamin was attached to a protein of apparent molecular weight 176 000.


Effect of vitamin D deficiency on sarcoplasmic reticulum function and troponin C concentration of rabbit skeletal muscle.

1. Weanling rabbits were made rachitic either by a vitamin D-deficient diet or by parenteral administration of ethane 1-hydroxy-1,1-diphosphonate (EHDP) in amounts sufficient in other species to block the formation of 1,25-dihydroxycholecalciferol [1,25-(OH)2D3]. 2. The uptake of calcium into the isolated sarcoplasmic reticulum from mixed striated quadriceps muscle, and the amount of troponin C (the calcium-binding component of the troponin complex) in relation to other proteins from the same muscle, were measured. 3. In muscle from animals made rachitic by a dietary deficiency of vitamin D, the rate of uptake of calcium by the sarcoplasmic reticulum and the troponin C concentration were both significantly less (P less than 0.02) than in control littermates. In EHDP-treated animals no significant differences from controls were found. 4. These results show that dietary deficiency of vitamin D in such animals can affect muscle physiology. Since no changes are found in animals made rachitic with EHDP, who presumably have a selective deficiency of 1,25-(OH)2D3, it is possible that the effect of vitamin D on muscle is mediated through metabolites other than 1,25-(OH)2D3 such as 25-hydroxycholecalciferol.


Comparison of 15N-labelled glycine, aspartate, valine and leucine for measurement of whole-body protein turnover.

1. Whole-body protein turnover was measured in rats by constant infusion of 15N-labelled glycine, aspartate, valine and leucine and measuring the enrichment of hepatic and renal urea and ammonia nitrogen. 2. The values obtained with [15N]glycine were comparable with values reported with methods based on different assumptions. 3. [15N]Aspartate gave rise to an increased enrichment of urea and ammonia and hence to lower protein-turnover rates. 4. [15N]Valine and [15N]leucine gave low enrichments of nitrogenous end products and hence to high protein-turnover rates. 5. All 15N-labelled amino acids are not equally suitable for measuring whole-body protein turnover by the end-product method. The relative amounts of 15N going to the end products can be predicted from the known individual metabolism of aspartate and the branched-chain amino acids.


Element and polychlorinated biphenyl deposition and effects in sheep fed cabbage grown on municipal sewage sludge.

Sheep were fed a diet containing 30 per cent (dry weight) cabbage which had been grown on municipal sewage sludge from Syracuse, NY for 77 days. Slight increases were found in the concnetration of nickel in kidney, copper in liver and cadmium in certain animal organs as compared to those in sheep fed a control ration containing cabbage grown on soil. Polychlorinated biphenyls were also found at higher levels in the sludge-grown cabbage and in the livers of the respectively fed sheep as compared to controls. An increase in liver weight, proliferation of hepatic smooth endoplasmic reticulum and various degenerative changes were noted in the livers of the sheep fed the sludge-grown cabbage ration as compared to the controls.


Object sorting by chimpanzees and monkeys.

Chimpanzees and two species of monkey were compared on sorting tasks. When the sets of objects differed in multiple ways, most chimpanzees learnt to sort whereas only a few monkeys performed better than chance (and then inconsistently). When the objects within a set differed in one or two respects, so that sorting by one principle entailed "unsorting" by one or more alternative principles, chimpanzees had greater difficulty. Two young animals did rapidly learn to sort by color and size ("unsorting" by size and shape or by color and shape). They did not sort pictorial material and had difficulty in sorting by shape if "unsorting" by both color and size. The sorting task differentiated between the cognitive capacities of chimpanzees and monkeys whereas other tasks did not.


The biology of the gonococcus.

Gonorrhea has been known since antiquity. Today, this disease is the most commonly reported infectious disease in the U.S. The natural environment of the etiological agent, Neisseria gonorrhoeae, is man. In this host, the organism usually parasitizes mucosal surfaces populated by columnar epithelial cells. Under certain conditions, the gonococcus may disseminate or spread to adjacent organs. The gonococcus is well adapted to its environment and is a successful parasite. Until recently, gonococci were uniformly sensitive to penicilin. However, a plasmid encoding beta-lactamase has been identified in some isolates. Most strains exhibit specific requirements for various amino acids, vitamins, purines, and pyrimidines. Only glucose, pyruvate, and lactate are utilized as sources of energy. Glucose is dissimilated by a combination of the Entner-Doudoroff and pentose phosphate pathways. A tricarboxylic acid cycle is also present and active under certain conditions. Structurally, the cell envelope of the gonococcus resembles that of a typical Gram-negative bacterium. Gonococci are highly autolytic, especially in older cultures or after depletion of the energy source. Autolysis is not due solely to peptidoglycan hydrolysis, but appears to involve a destabilization of the outer membrane as well. Cell surface components such as pili, lipopolysaccharide, outer membrane proteins, and a capsule are associated with the virulence and pathogenicity of this organism.


Very large analogy of chromosome banding between Cebus capucinus (Platyrrhini) and man.

The karyotype of a platyrrhine, Cebus capucinus, is compared with that of man and other catarrhines. It was found that a large degree of analogy of chromosome banding exists among the different species. A common origin for the platyrrhines and the catarrhines can thus be deduced. Furthermore, it becomes possible to reconstruct, to a great extent, the karyotype of the last common ancestor of all of the simians.


Chromosomes of Lemuriformes. V. A comparison of the karyotypes of Cheirogaleus medius and Lemur fulvus fulvus.

In this report we compare the karyotype of Lemus fulvus fulvus (2n=60) with that of Cheirogaleus medius (2n=66), a species thought to retain the ancestral lemur karyotype. A culture technique was designed specifically for lemur lymphocytes to facilitate description of the complete karyotypes using G--banding, C-banding, and Ag-AS staining for nucleolus organizer regions (NOR's). Different G-banding patterns in three chromosome pairs and different NOR-bearing chromosomes between the two species, as well as additional chromosomes and interstitial C-bands in C. medius, suggest that the chromosome complement of C. medius may not perfectly reflect the ancestral morphology. However, allowing for a Robertsonian centric fusion and a pericentric inversion, the G-banding patterns of 27 of the 32 autosomal pairs of C. medius are indistinguishable from those of L. fulvus fulvus. This constitutes strong justification for assigning these chromosomes to the ancestral lemur karyotype.


In vitro efficacy of Bay k 4999, a new ureido-penicillin, in combination with aminoglycosides against Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae and Proteus strains.

The in vitro effects of Bay k 4999 in combination with gentamicin, tobramycin, amikacin sisomicin and netilmicin in bacteriostatic (MIC) and bactericidal (MBC) concentrations were compared using the checkerboard dilution technique against 20 different strains of Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae and indole-positive-negative Proteus species. On average 63% of Bay k 4999-aminoglycoside (AG) combinations inhibited Pseudomonas, Proteus and Klebsiella strains additively and/or synergistically in bacteriostatic as well as in bactericidal concentrations as compared to only 14% additive or synergistic activity on E. coli. 35% of the combinations tested proved to be synergistic in K. pneumoniae, 20% in Proteus, 13% in Pseudomonas, but only 5% in E. coli. No significant differences between various Bay k 4999-AG combination effects could be demonstrated.


A comparison of the in vitro activity of tobramycin and gentamicin against 6,042 clinical isolates.

6,042 strains of Staphylococcus aureus, Streptococcus faecalis, Proteus mirabilis, Escherichia coli, Klebsiella sp. and Pseudomonas sp. were tested against tobramycin and gentamicin. Among all species studies, except Pseudomonas sp, gentamicin turned out to be slightly superior to tobramycin.


[Bacteriological findings in patients with cholelithiasis and duodenal diverticuli].

In a prospective study of 70 patients with duodenal diverticula and calculous biliary tract disease, bacteriocholia with typical intestinal bacteria was found in 69%, whereas in cholelithiasis without duodenal diverticula this complication was present in 30% of the cases. With growing distance between duodenal diverticula and the papilla of Vater the rate of bacteriocholia decreases. Rising age of patients results in augmentation of bacteriocholia. Bacteriocholia on the basis of duodenal diverticula seems to present an additional pathogenetic factor for inflammatory biliary tract disease in 69% and for pancreatitis in 20% of the cases. Consequently the pathologic value of diverticulas close to the papilla of Vater is more prominent than reported so far.


Biosynthesis of prostaglandins.

The generation of prostaglandins is catalyzed by a membrane-bound multienzyme complex. The first reaction of the biosynthetic sequence is the generation (by the enzyme 'fatty acid cyclooxygenase') of prostaglandin endoperoxides, a reaction which involves the incorporation of two moles of oxygen: this reaction probably proceeds by an ene reaction rather than a free-radical mechanism. After biosynthesis the endoperoxides can be metabolized in various ways depending on the cell-type. For example, in platelets they may be transformed into non-prostanoid compounds called thromboxanes, whereas vascular endothelium and many other tissues generate another derivative, prostacyclin. In other tissues, the 'classical' prostaglandins E, F or D may be generated. Some products of the cyclooxygenase (e.g. hydroperoxides, malonaldehyde) may have a direct toxic action on cells: prostaglandins themselves do not, but some types (especially those of the E series) are probably responsible for many of the clinical signs and symptoms of inflammation.


[Basic conditions for electromyographic examination. Part I: EMG, nerve conduction velocity and test of motor end-plate function (author's transl)].

The purpose of the following contribution is to give a survey about the basic conditions of electromyographic examination. The necessary apparatus will be described as well as the course of examination. Moreover the parameter for the evaluation will be explained together with exogenous falsifying influences. This contribution is thought to be a guide for orientation to improve electromyographic recording and evaluation.


[The sex differences in the normal resting EEG of young adults (author's transl)].

In order to investigate the sex differences in the normal resting EEG, 488 healthy students (290 males and 198 females) were examined using a computer program for time domain analysis. Women show on the average more beta-waves, less alpha-waves, higher beta-amplitudes, higher average frequencies, and other signs of a higher average level of arousal. However, the total variability of all examined EEG parameters is present in both sexes. There are some hints that laterality of EEG in precentral leads could be more pronounced in males than in females.


[The short periodicity of paroxysmal complex waves in electroencephalographic and clinical course of Creutzfeldt-Jakob disease (author's transl)].

The Creutzfeldt-Jakob disease reveals striking bioelectrical and clinical features which provide indirect confirmation of actions in cortical, subcortical and brainstem neuronal systems determining the type, the stage and the fatal limits of the course. The periodic paroxysmal complex waves in EEG, the signs of motor, vigilance and respiratory impairment and the site of lesions are delineated in a case of cortico-striato-cerebellar type and compared with other cases in literature. Other forms of electroencephalographic periodicity in different clinical and experimental conditions are discussed. The period dynamics are referred to the course of disease and investigated under the influence of anticonvulsive drugs.


[Electroencephalographic changes in coma due to hepatic insufficiency with porto-caval shunt (author's transl)].

EEG studies were done in 19 patients comatose due to severe hepatic insufficiency associated with porto-caval shunting. The neurological deficits were graded in 5 stages and related to the abnormal EEG-pattern observed in 38 records. With deepening of coma, alpha-frequencies were gradually replaced by theta- and delta-activities. The frequency of theta- and delta-runs decreased with neurological deterioration. Triphasic waves (typical and/or atypical) were seen in 26% of the records. Typical triphasic waves were found to be limited to stage 3 of coma, whereas atypical forms appeared in earlier and later stages. After infusions of L-Valin, a transient improvement and disappearance of triphasic waves was observed, but no significant effect could be established on the final course of the disease. Sleep-like potentials and the reactivity to external stimuli decreased in deep stages of coma. Severe EEG abnormalities were correlated to low cholinesterase and high lactid acid blood level, but no correlation was found to the level of ammonia.


[Epilepsy and psychoses (from the viewpoint of a neurosurgical service)].

The experience of a neurosurgical unit devoted to the surgical therapy of the epilepsies is only based on very peculiar patients. However, relationship between a given epilepsy and associated psychiatric disorders are more easily understood because of more extensive explorations. Bilateral temporal epilepsies involving the limbic system on the one hand, bilateral frontal epilepsies on the other one, and P.M. status which may be paralleled, make these patients more susceptible to acute mental confusions, to acute thymic disorders, to delirious attacks. Direct relations between chronic psychosis and severe epilepsies are much more uncertain.


[Lithium and thyroid function. Significance of the TRH test in the diagnosis of lithium-induced thyroid dysfunction].

The treatment by lithium is known to involve certain endocrine complications. Those concerning the thyroid function, with risk of a frank hypothyroidy, are the most important. Aiming to appreciate the frequence and the intensivity of the endocrine effects of lithium, the thyroid parameters and the steady state of the hypothalamo-pituitary-thyroid axis were tested using the TRH test in 52 patients with maniaco-depressive psychosis with special attention to TSH, prolactin and growth hormone: 24 out of them were treated for 1 month to 6 years by lithium; the 28 others were considered as controls. The lithium treatment involves a decrease in the free thyroxine index (1.78 +/- 0.09 vs 2.16 +/- 0.09; p less than 0.01), an increase in the mean baseline TSH level (5.80 +/- 1.49 vs 2.70 +/- 0.24 microU/ml; p less than 0.05) and a noteworthy increase in the TSH responsiveness to TRH (22.7 +/- 2.14 vs 9.75 +/- 1.63 microU/ml; p less than 0.005). The TSH supranormal responses were neither correlated with the length of the treatment nor with the age of the patients. They appear as the consequence of a decrease in the thyroidal hormone secretion. The basal and stimulated prolactinemias remain comparable in the two groups of patients and no response of growth hormone occured after TRH. The TRH test must be considered as a useful complement for the surveillance of the patients treated with lithium because it permits to diagnose early the lithio-induced thyroid dysfunction.


Androgen and estrogen metabolism in male rhesus monkeys.

Using constant infusions of 3H-labeled androgens and 14C-labeled estrogens, the dynamics of androgen and estrogen metabolism have been studied in male rhesus monkeys. The mean (+/-) MCR, as measured in whole blood, for testosterone (T; 140 +/- 10 liters/day) was significantly smaller than the MCRs for androstenedione (A), estrone (E1), and estradiol (E2; 650 +/- 8, 1010 +/- 160, and 500 +/- 20 liters/day, respectively). These findings are compatible with the specific globulin binding of T in this species. The mean blood productions were 0.42 +/- 0.11 and 0.55 +/- 0.04 mg/day for A and T and 40 +/- 16 and 13 +/- micrograms/day for E1 and E2, respectively. The interconversions of the androgens [fractional conversion rates of A to T ([rho]A,TBB) and T to A ([rho]T,ABB)] were not different but were lower than the interconversions of the estrogens ([rho]E1,E2BB and [rho]E2,E1BB). The mean [rho]T,E2BB values were 0.015 +/- 0.001 and 0.0024 +/- 0.0003, which are remarkably similiar to those values in men. Administrations of PMS gonadotropin did not alter any measured values of androgen or estrogen metabolism, hCG resulted in a decrease in the conversion ratio of A to T, T administration increased the MCR of T, and estrogen administration resulted in no changes. The aromatization rate remained unaffected by all treatments.


Human chorionic gonadotropin in BeWo trophoblastic cells after 12 years in continuous culture: retention of intact human chorionic gonadotropin secretion in mechanically versus enzyme-dispersed cells.

Gel filtration and radioimmunological studies were used to demonstrate that BeWo malignant trophoblastic cells, subcultured for more than 12 yr as roller tube colonies using dissection by scalpel, retained the ability to secrete large quantities of intact hCG. By contrast, BeWo cells that had been subcultured weekly for 12 hr using proteolytic enzyme dispersion lost their ability to secrete intact hCG. Rather, a large form of hCG-alpha was the major secretory product, along with lesser amounts of heterogeneous low molecular weight forms of hCG-beta. The gradual loss of hormone secretory ability and fidelity of hormone product by cells in continuous culture is a well known phenomenon. Our results suggest that the habitual use of trypsin-EDTA to subculture cells may accelerate this process.


Experimental anticonvulsant cinromide in monkey model: preliminary efficacy.

Cinromide (3 brono-N-ethylcinnamide), an experimental anticonvulsant (Burroughs-Wellcome Pharmaceutical Co.), was given a preliminary evaluation in our alumina-gel monkey model. The parent drug has a biological half-life in monkey of 1-2 hr and its active metabolite, 3-bromocinnamide, a half-life of 4-6 hr. In phase 1, 6 chronically epileptic monkeys, with focal motor and secondarily generalized tonic-clonic seizures, received the drug in a vehicle of 65% polyethylene glycol 400 (PEG) by constant-rate intravenous infusion followed by baseline days of saline only and PEG only. Three different concentrations of Cinromide (12, 24, and 36 mg/ml/hr) were administered, respectively, to achieve mean steady state plasma levels of approximately 5, 10 and 20 micrograms/ml of the metabolite (0.5 to 5.0 micrograms/ml of the parent drug). In phase 2, Cinromide was administered for 7 days at the middle concentration to all monkeys. Baseline periods similar to those of phase 1 were used as controls. The data tentatively suggest that Cinromide is efficacious in the monkey model at a plasma concentration range of 7-14 micrograms/ml of the metabolite. With the exception of one animal, no secondarily generalized seizures were exhibited during drug administration (but were evident in the baseline periods), and EEG bursting decreased significantly in several monkeys. Minimal side effects were manifested at these plasma levels but withdrawal seizures were evinced with cessation of the drug. Further evaluation of Cinromide by gastric administration in our animal model is planned.


The influence of cannabidiol and delta 9-tetrahydrocannabinol on cobalt epilepsy in rats.

The mechanisms of the anticonvulsant activity of cannabidiol (CBD) and the central excitation of delta 9-tetrahydrocannabinol (delta 9-THC) were investigated electrophysiologically with conscious, unrestrained cobalt epileptic rats. The well-known antiepileptics, trimethadione (TMO), ethosuximide (ESM), and phenytoin (PHT), were included as reference drugs. Direct measurements were made of spontaneously firing, epileptic potentials from a primary focus on the parietal cortex and convulsions were monitored visually. ESM and TMO decreased the frequency of focal potentials, but PHT and CBD exerted no such effect. Although CBD did not suppress the focal abnormality, it did abolish jaw and limb clonus; in contrast, delta 9-THC markedly increased the frequency of focal potentials, evoked generalized bursts of polyspikes, and produced frank convlusions. 11-OH-delta 9-THC, the major metabolite of delta 9-THC, displayed only one of the excitatory properties of the parent compound: production of bursts of polyspikes. In contrast to delta 9-THC and its 11-OH metabolite, CBD, even in very high doses, did not induce any excitatory effects or convulsions. The present study provides the first evidence that CBD exerts anticonvulsant activity against the motor manifestations of a focal epilepsy, and that the mechanism of the effect may involve a depression of seizure generation or spread in the CNS.


Relative significance of isolated infantile convulsions as a primary cause of focal epilepsy.

Isolated infantile convulsions were noted in 206 patients (13%) of 1,572 patients who underwent cortical resection for medically refractory focal epilepsy at the Montreal Neurological Institute over the period 1928 through 1977. In 59 patients (29% of the 206), the isolated infantile convulsion was associated with an identifiable febrile systemic or neural illness. In 54 patients (26%), there was a definite history of cerebral birth injury in addition to the isolated infantile convulsions. Twenty patients (10%) had some other potential etiological factor for the later development of seizures, such as postnatal head injury, Sturge-Weber syndrome, etc., in addition to the isolated infantile convulsions. In 95 patients, 6% of the total 1,572 patient series and 41% of those with isolated infantile convulsions, the latter was the sole apparent cause for the later development of recurring seizures. Although this was the case in a small but significant percentage in this series of patients, more frequently, the isolated infantile convulsion represents a manifestation of invasion of the brain by an infectious agent of a systemic or brain disease, or is a harbinger of the later development of a seizure tendency due to some other preexisting etiological factor.


Electroencephalographic and behavioral effects of a GABA agonist (muscimol) on photosensitive epilepsy in the baboon, papio papio.

Muscimol, 0.25-1.0 mg/kg, i.v., was administered acutely to 4 adolescent baboons, Papio papio, that show photically induced epilepsy. On the EEG, slowing of background rhythms was associated with the appearance of spikes, polyspikes, and recurring symmetrical spike-wave complexes. These changes were maximal 0.5-2 hr after muscimol injection. Regular testing with intermittent light stimulation showed either no change from control responses or a more severe epileptiform EEG 0.1-3 hr after muscimol. Photically induced myoclonus was not modified by muscimol. Despite its GABA-abonist properties, muscimol is not an effective anticonvulsant.


Endocrine factors and glucose metabolism during prolonged seizures in baboons.

Changes in plasma glucose, nonesterified fatty acids, insulin, glucagon, cortisol, growth hormone, and prolactin have been studied in baboons during the course of generalized epileptic seizures induced by intravenous bicuculline. Plasma glucose rose to a peak at 25 min but fell to hypoglycemic levels after 60 min of seizure activity. This hypoglycemia was accompanied by a marked elevation in plasma insulin. Plasma glucagon rose to a peak at 14 min, then returned to normal. Plasma growth hormone levels were elevated after 60 min of seizure activity. Plasma prolactin and cortisol levels also rose during the seizure. These changes result from sequential interaction of (1) autonomic activation at seizure onset, (2) spread of neuronal activity to the hypothalamus leading to the liberation of releasing factors, and (3) indirect physiologic consequences of seizure activity.


Purification, characterization and localization of serine protease of Morris hepatoma 8999.

1. A serine protease of hepatoma 8999, isolated in the mitochondrial fraction, was purified and crystallized. The purified enzyme was apparently homogeneous on ultracentrifugal analysis and polyacrylamide disc gel electrophoresis. The ratio of absorbance at 280 nm and 260 nm, A280/A260, was 1.90 and its absorption coefficient, A280 1% was 10.5 cm-1 estimated from dry weight measurements. Its S20, w value was 2.23 S and its molecular weight was estimated to be 24000 +/- 1000. The enzyme contained twice as much lysine, arginine and histidine as chymotrypsinogen did, but had a very similar amino acid composition to serine protease from skeletal muscle. Its isoelectric point was pH 10.6. 2. The substrate specificity of the enzyme was the same as that of chymotrypsin A. Its Km and kcat values for N-acetyl-L-tyrosine ethyl ester, N-acetyl-L-phenylalanine ethyl ester and N-acetyl-L-tryptophan ethyl ester were 0.35 mM and 10.69 s-1, 0.38 mM and 10.7 s-1, and 0.11 mM and 11.8 s-1, respectively. Its activity was completely inhibited by phenylmethylsulfonyl fluoride and partially inhibited with tosylphenylalanine chloromethyl ketone. 3. The enzyme was shown to be located in different granules from the intracellular particules (light and heavy mitochondrial fraction) by sucrose density gradient centrifugation, and it was stained in mast cells of the hepatoma 8999 by the immunofluorescent technique. 4. Serine protease is present in different amounts in various organs of rat and the enzyme from hepatoma 8999 gave a single band that fused completely with those of the enzymes from skeletal muscle, heart, liver and kidney, respectively, on Ouchterlony double-diffusion analysis using antiserum to the crystalline enzyme of hepatoma 8999, but the enzyme from small intestine did not react with the antiserum.


Translation in vitro of Tetrahymena pyriformis polyadenylated mRNA. Identification of tubulin amongst the translated products and demonstration of its heterogeneity.

The capacity of poly(A)-containing RNA of the protozoan ciliate Tetrahymena pyriformis to direct the synthesis of proteins in vitro has been tested using two cell-free systems: a wheat germ extract and a rabbit reticulocyte lysate. The results obtained with these two systems are compared and the identification of alpha and beta tubulins among the products of protein synthesis in vitro, after separation by one-dimensional and two-dimensional electrophoresis, is described. By isoelectric focusing in polyacrylamide gels, each species of tubulin is resolved into several bands, suggesting that the main subunits are more heterogeneous than has been generally described. Poly(A)-containing RNA has also been fractionated on a 70% formamide/sucrose gradient and it is shown that alpha and beta tubulins are coded by separate mRNAs.


Nuclear-magnetic-resonance study of the conformation of a dinucleotide in solution using the lanthanide probe method.

The conformation of the dinucleotide adenylyl(3'-5')adenosine 2'-phosphate (ApA2'p) in aqueous solution at different pH values and temperatures has been studied using the lanthanide(III) ethylenodiaminetetraacetate(EDTA) 1:1 complexes as shift and relaxation probes. The conformational analysis, based on mixing different conformations in fast exchange within the nuclear magnetic resonance time scale, agrees well with the results from vicinal coupling constants and dimerization shifts obtained for the metal-free system. The dinucleotide exists in a temperature-dependent and pH-dependent conformational equilibrium between unstacked and base-stacked forms. At neutral pH and low temperature, the stacked form predominates, and it is a predominantly right helical structure, characterized as an anti, 3E, g-, g- g-, g'g', gg, 3E, anti conformation. This situation contrasts with adenylyl(3'-5')adenosine (ApA), where both right and left helices contribute to the stacked form. The nucleotidyl units of the unstacked form of ApA2'p have average conformations which are very similar to those of the corresponding mononucleotides in similar conditions.