Bladder cancer as seen in giant histologic sections. Giant sections from cystectomy specimens in 45 cases of bladder cancer were examined microscopically after en bloc fixation and processing. There were 35 transitional cell, seven squamous cell, and three mixed transitional cell and squamous carcinomas. Broad front invasion was associated with papillary and superficial tumors while tentacular invasion was associated with solid tumors and a generally poorer prognosis. Carcinoma in situ merged with the invasive cancer in 33 cases, and neoplasia in these cases tended to be multifocal. In 10 cases there was no carcinoma in situ next to the invasive lesion, and the cancer was unifocal. These findings support the concept that there may be two pathogenetic types of bladder cancer, one arising in an extensive field of abnormal epithelium and one arising in a focal area of abnormality. The findings also underscore the importance in clinical management of selected mucosal biopsies adjacent to the site of any visible bladder tumors.


An overview: antitumor immunity in breast cancer assayed by tube leukocyte adherence inhibition. The adherence to glass of human peripheral blood leukocytes (PBL) incubated with tumor antigen in vitro, is specifically inhibited if the PBL are sensitized to the antigen. The presence of leukocyte adherence inhibition (LAI) to tumor extracts indicates the presence of systemic antitumor immunity. By the tube leukocyte adherence inhibition assay (tube LAI), it was shown that 85% (191 of 223) Stage I and II, 45% (15 of 34) Stage III and 29% (30 of 103) Stage IV breast cancer patients had LAI reactivity. LAI responsiveness diminished with an increased tumor burden and most patients with advanced cancer exhibited no LAI reactivity. When LAI reactivity was monitored for 1 to 6 months after surgery, 13 of 25 Stage I and II breast cancer patients were negative on the first repeat assay. In general, 7 months after mastectomy most patients clinically free of cancer showed no LAI reactivity. Of thirty-five patients tested between 7 and 18 months after mastectomy, 6 were positive and 4 of the positives had local recurrence. The phenomenon of tube LAI appears to be mediated by monocytes armed with cytophilic antitumor antibody. The serum of patients whose leukocytes responded in the tube LAI assay had free cytophilic antitumor antibody that "armed" or sensitized normal leukocytes to respond in the LAI assay. Serum arming paralleled leukocyte reactivity before and after surgery. Patients with advanced cancer whose leukocytes failed to react in the LAI assay had serum blocking factors (excess tumor antigen) that abrogated the LAI reactivity of leukocytes from reactive patients.


Randomized sequential hormonal therapy vs adrenalectomy for metastatic breast carcinoma. One hundred sixty-one postmenopausal and 65 premenopausal women, a total of 226 patients with metastatic breast carcinoma, were included in this randomized study to evaluate the merits of adrenalectomy as the primary mode of therapy as compared to the customary sequential hormonal manipulation. The 145 evaluable postmenopausal patients were randomized as follows: (1) primary additive hormone therapy first followed by adrenalectomy and (2) primary adrenalectomy followed by chemotherapy and/or additive hormone therapy. When 76 patients in group 1 were compared with 70 patients in group 2 regarding their survival time, there was no essential difference, but the response rate was 20% vs 38.6%, a significant difference. The 55 evaluable premenopausal women were randomized into two groups: (1) oophorectomy followed by adrenalectomy; (2) adrenalectomy-oophorectomy as primary mode of therapy. The response rate in group 1 was 17.4% as compared with 41.9% in group 2, but again there was no difference in the survival time among these two groups. When sequential hormonal manipulation was utilized, only one-third of these patients were subjected to adrenalectomy because of their rapidly deteriorating condition. Adrenalectomy performed as a secondary procedure showed a lower response rate but the total survival time was comparable with primary adrenalectomy patients.


Effect of dose, route, and schedule of BCG on antibody titer and survival of murine skin grafts across a weak histocompatibility barrier. Skin was transplanted from male to female C57BL/6JRC mice. Bacillus Calmette-Guérin (BCG) was administered once subcutaneously, intramuscularly, or intraperitoneally either 14 days prior to grafting or on the day of grafting in a dose of 5 X 10(5), 5 X 10(6), or 5 X 10(7) microorganisms. Serum antibody to BCG was assayed using complement fixation. There was a linear inverse relation between skin graft rejection and dose of BCG (p less than .01) such that the lowest dose of BCG was the most effective. The IM and SQ routes were more effective than the IP route. Schedule did not have significant effect on skin graft survival. Time to appearance of antibody correlated directly with dose. The prolonged survival of skin grafts combined with early appearance of antibody to BCG suggested that high doses of BCG could result in antigenic competition.


Growth of human tumors in lethally irradiated mice reconstituted with syngeneic fetal liver cells. Transplantation into lethally irradiated mice of hematopoietic and lymphoid cells from immature donors which hypothetically will not mount a cell mediated attack against simultaneously inoculated human tumor cells has resulted in tumor engraftment and growth in long-term surviving radiation chimeras. Twenty-four hours after lethal irradiation, A or CBA mice were given iv injections of 2 X 10(7) fetal liver cells from syngeneic donors of 14, 16, or 18 days of embryonation and sc injections of 1, 3, or 6 X 10(6) human choriocarcinoma (C-1, C-2, and C-3) cells or human breast carcinoma (B-1) cells that had been maintained in culture. Palpable tumors greater than or equal to 5 mm were noted in 18/22 mice injected with C-1, 9/16 with C-2, 10/10 with C-3, and 18/30 with B-1. Tumors of 17 (31%) of mice remained palpable until death of the animal or until termination of the experiment 100 days post inoculation. Histologic study of autopsy specimens revealed malignant tumors with occasional pulmonary metastases. Human chorionic gonadotropin was found in the serum of mice that received choriocarcinoma cells.


Rhabdomyosarcoma: response with chemotherapy prior to radiation in patients with gross residual disease. One hundred and ten Group III patients enrolled in the IRS Study have been reviewed. Of these, 39 are fully and 56 are partially evaluable. Combining the data for both groups 77/95 (81%) have shown complete or partial response/no measurable disease and this has been maintained in 53/95 (56%). No patient received premature RT for local progression during the time of pre-radiation chemotherapy. However, 15/95 patients have shown local recurrence or direct extension to neighboring tissues; 13/15 could be reviewed. Only 3/13 have recurred at a time related to the delay in starting radiation. Overall survival for Group III patients is 72%-85% (Regimen F and Regimen E respectively).


Hyperbaric oxygen in radiation therapy.

Hyperbaric oxygen in radiation therapy. The importance of oxygen with low LET radiations has been established beyond any doubt in many different systems, both plant and animal. While some studies, especially head and neck tumors, are impressive, it has not been demonstrated unequivocally that radiation under hyperbaric conditions is superior to well fractionated, well conceived, conventional radiotherapy. Any resulting gain in survival from the addition of hyperbaric oxygen will be limited, especially with more advanced stages of disease. Well controlled studies, especially with earlier stage disease, are still necessary. It would be worthwhile to undertake such trials, especially with tumors of the head and neck constituting the most promising site of study, as others have noted, since even a 5% to 10% improvement in survival would mean many lives saved. Continued trials with hyperbaric oxygen, oxygen in other forms, neutrons and other particles, and radiation sensitizing drugs are all justified in an attempt to overcome the oxygen effect on human tumors.


Hyperthermic potentiation: biological aspects and applications to radiation therapy. Experimental studies have provided evidence that hyperthermia may be an effective agent, either alone or in combination with ionizing radiation, in the treatment of cancer. Results have shown that temperatures in the range of 42 degrees to 45 degrees C: 1) are cytotoxic, with cell lethality showing little or no dependence on levels of oxygenation; 2) inhibit the recovery of cells from sub-lethal and potentially lethal radiation damage while enhancing the levels of lethal damage; and 3) may be combined with x-irradiation in a manner to improve therapeutic ratios. The observed interaction between hyperthermia and x-rays may in part be due to differences in the Age Response Functions and reassortment of cycling cells to these two agents. Hyperthermia may also greatly change repopulation and re-oxygenation parameters in irradiated tumor and normal tissue volumes. An overall consideration of these and other factors is essential in the design of optimal schedules of combined hyperthermia and x-irradiation treatments in the management of malignant disease.


Radiotherapy in the postoperative treatment of operable cancer of the breast. Part II. A re-examination of Stjernswärd's application of the Mantel-Haenszel statistical method. Evaluation of the effect of the radiation on immune response and suggestions for postoperative radiotherapy. A re-examination of the statistical methods used to develop the claim that radiation postoperatively decreases survival was carried out. This study demonstrates that a) when correct figures are used, statistical significance is not demonstrated, i.e. there is not statistically significant evidence that irradiation of the peripheral lymphatics and/or chest wall following mastectomy has a deleterious effect on survival, b) the use of the Mantel-Haenszel procedure was inappropriate for the purpose for which it was used. A survey of the available evidence fails to substatiate the claim that the immunological system effects of local irradiation relate in any way to decreased survival. Recommendations for the use of irradiation postoperatively in the treatment of breast cancer are made.


Clinical trials of adjuvant radiation therapy for breast cancer. Years ago, radiation therapists conducted randomized clinical trials which showed that regional irradiation significantly reduces local recurrence of breast cancer but does not often affect life span. No new findings challenge these conclusions. Controlled trials in general have lost some credibility, however, as it has become more widely recognized that they are very vulnerable to fallacious reasoning and to simple human error. There is serious concern that some physicians may be unwisely abandoning well tested treatment methods because of the premature dissemination of early results of some trials. Still, many important questions remain that can only be answered by clinical trials, some including radiation therapy. This paper suggests that randomization schemes be incorporated into "routine" medical practice, although only after the most careful deliberation, in order to hasten fuller understanding of old therapeutic methods as well as new ones.


Endometrial cancer: therapeutic decision and the staging process in "early" disease. An all inclusive, widely accepted system for correlation of indices of pathophysiology in endometrial cancer with a spectrum of therapeutic management has yet to be developed. Improved understanding of tumor growth should lead to more logical, individualized treatment especially in terms of irradiation. To support these philosophies a brief review of past reports and studies, especially the Endometrial Adjuvant Study is provided. From an analysis of 574 patients in this study, it is apparent that prognostic factors could be separated as major, differentiation, and tumor penetration and minor, number of capsules of radium and depth of uterus. A pilot study under the Gynecologic Oncology Group suggests the correlation of the major factors with lymph node involvement. Since depth of penetration and lymph node involvement are most accurately determined by surgery and pathology, surgical staging is suggested as a guide for therapeutic decision.


Chemical modification of radiation effects. A number of powerful chemical compounds that modify radiation effects have been discovered and tested both in the laboratory and clinically over the past 25 years. There are four major classes of compounds: aminothiol radio-protectors which act on well vascularized euoxic cells and concentrate in tissues such as skin, gut and marrow; nitromidazole radiosensitizers which act on hypoxic tumor cells; pyrimidine analogues which are incorporated into the DNA of cycling cells and cause radiosensitization; and cancer themotherapy agents which, in addition to their ability to kill tumor cells directly, also may sensitize tumor and normal cells to radiation. The mechanism of action, experimental activity, and clinical results or the potential for each of these agents are reviewed.


Relationship of glucagon-somatostatin and gastrin-somatostatin cells in the stomach of the monkey. The stomach of the monkey Tupaia belangeri was investigated by serial sections utilizing the indirect immunoperoxidase reaction to demonstrate the distribution of glucagon, gastrin and somatostatin immunoreactive cells. A striking topographical distribution was found. Glucagon and somatostatin immunoreactive cells were located in the upper parts, whereas gastrin and somatostatin immunoreactive cells were situated in the lower parts of the stomach. The remaining regions of the stomach did not contain cells immunoreactive to the antisera applied. Similarly, the ultrastructural study revealed the same distribution of endocrine cell types identified as A-cells, D-cells, and G-cells. Thus, there may be a glucagon-somatostatin area in the upper part and a gastrin-somatostatin endocrine surface in the lower part of the stomach. This spatial relationship of the endocrine cells suggests a functional cell interaction between glucagon and somatostatin cells in the cranial stomach and between gastrin and somatostatin in the caudal parts of the stomach.


Electron dense plaques (calcium binding sites) in the plasma membrane of the endocrine cells in rodent pancreatic islets and parathyroid glands. Small electron dense plaques (EDP) were found in the plasma membrane of pancreatic islet A-, B- and D-cells and parathyroid chief cells of mice and gerbils. The identification of the EDP was facilitated by the use of special fixation techniques. The EDP may represent sites of calcium binding in the cell membranes.


Growth characteristics of postnatal rat adrenal medulla in culture. A study correlating phase contrast, microcinematographic, histochemical, and electron microscopical observations. Explants and enzyme-dispersed cells of adrenal medulla from 10-12 day old rats were studied in culture for up to 3 weeks. Adrenomedullary chromaffin cells, nerve cells and satellite cells were clearly discernible. The nerve cells were few in number and did not show catecholaminespecific fluorescence. Chromaffin cells stored catecholamines, as judged by the Falck and Hillarp method, in varying amounts decreasing with age of the cultures and the distance from the explants. Exocytosis profiles observed with the electron microscope suggested that cultured chromaffin cells also released catecholamines. Moreover, the cells formed processes and frequently migrated into the outgrowth. After 6 days in culture, the great majority of chromaffin cells stored noradrenaline as revealed by electron microscopy with few adrenaline-storing cells being visible. Granular vesicles (approximately 80-240 nm in diameter) with cores of different electron densities were occasionally present in the same cell suggesting the occurrence of mixtures of primary and secondary amines. Apart from "chromaffin" granules, small clear and dense-cored vesicles (approximately 40-60 nm) were found both in the somata and cell processes. Chromaffin cells and their processes were often closely apposed and occasionally formed specialized attachment zones. As a whole, chromaffin cells in culture resembled small granule-containing cells in sympathetic ganglia. 0.5 mM dbcAMP prevented dedifferentiation of chromaffin cells as judged by the lack of processes, the size and amount of "chromaffin" granules and the high number of adrenaline-storing cells present after 6 days in culture. NGF caused a striking increase in the number of axons growing out from explants.


Localization of somatostatin-like immunoreactivity in the pancreatic islets of the hagfish, Myxine glutinosa and the lamprey Lampetra fluviatilis. Somatostatin-like immunofluorescence has been found by immunostaining in cells of the bile duct mucosa and pancreatic islet parenchyma of the hagfish, Myxine glutinosa, and the islet lobules of the lamprey, Lampetra fluviatilis.


Immunocytochemical demonstration of thehypothalamo-hypophysial vasotocinergic system of Lampetra fluviatilis. With the use of the unlabeled antibody peroxidase-antiperoxidase (PAP) technique at the light microscopic level, it was shown that the preoptico-hypophysial neurosecretory system of the adult migrating Lampetra fluviatilis is a vasotocinergic system. It does not synthesize vasopressin. The results are entirely consistent with earlier chromatographic and pharmacological indications thatit produces little or no oxytocin-like peptide hormone. In the adenohypophysis, immunoreactive neurohypophysial peptidergic fibres are absent.


Distinction between smooth muscle, fibroblasts and endothelial cells in culture by the use of fluoresceinated antibodies against smooth muscle actin. FITC-labelled antibodies against native actin from chicken gizzard smooth muscle (Gröschel-Stewart et al., 1976) have been used to stain cultures of guinea-pig vas deferens and taenia coli, rabbit thoracic aorta, rat ventricle and chick skeletal muscle. The I-band of myofibrils of cardiac muscle cells and skeletal muscle myotubes stains intensely. In isolated smooth muscle cells, the staining is located exclusively on long, straight, non-interrupted fibrils which almost fill the cell. Smooth muscle cells which have undergone morphological "dedifferentiation" to resemble fibroblasts with both phase-contrast microscopy and electronmicroscopy still stain intensely with the actin antibody. In those muscle cultures which contain some fibroblasts or endothelial cells, the non-muscle cells are not stained with the actin antibody even when the reactions are carried out at 37 degrees C for 1 h or after glycerination. Prefusion skeletal muscle myoblasts also do not stain with this antibody. It is concluded that the actin antibody described in this report is directed against a particular sequence of amino acids in muscle actin which is not homologous with non-muscle actin. The usefulness of this antibody in determining the origin of cells in certain pathological conditions such as atherosclerosis is discussed.


Specific immunoelectronmicroscopic localization of vasopressin and oxytocin in the neurohypophysis of the rat. An immunoelectronmicroscopic method for the specific localization of neurohypophyseal hormones was developed in neurohypophyses of Wistar and Brattleboro rats, the latter strain being homozygous for diabetes insipidus. If the proper precautions were omitted, a marked cross reactivity between anti-vasopressin and anti-oxytocin preparations was found. Cross reaction of an anti-vasopressin plasma with oxytocin, at a dilution of less than 1:1600, resulted in electron density of all granules within neurosecretory fibres of the Brattleboro and Wistar neurohypophyses. However, this cross reactivity could be eliminated either by sufficient dilution of the anti-plasma, or by its purification. Purification of the antibodies was performed by absorption to agarose beads coated with the cross reacting component. Upon incubation with anti-vasopressin (diluted unpurified 1:1600 or purified 1:80) and unpurified anti-oxytocin (1:400) plasma, sections of a Wistar neurohypophysis revealed two types of neurosecretory fibres, containing either electron dense or lucent granules. Oxytocin and vasopressin containing neurosecretory fibres were found as clusters in the neurohypophysis. The specificity of both unpurified anti-vasopressin (1:1600) and anti-oxytocin (1:400) plasma was confirmed on serial sections of a Wistar neurohypophysis, alternately incubated with the solutions of the two antibodies. These data prove that the one-cell-one-hormone hypothesis holds true for the hypothalamic-neurohypophyseal system.


The hypothalamo-choroidal tract. I. Immunohistochemical demonstration of neurophysin pathways to telencephalic choroid plexuses and cerebrospinal fluid. Neurosecretory pathways were examined in normal male rats by the use of the immunoperoxidase technique for the localization of neurophysin in Bouin-fixed, deparaffinized sections. Using this technique two projections of extrahypothalamic neuorosecretory fibers can be traced to the sites of origin of the choid plexuses of both horns of the lateral ventricles. Neurophysin-containing axons originating primarily from the paraventricular field course dorsolaterad to enter the choroid fissure of the dorsal horn. A caudally direced group of fibers course ventrolaterad to enter the ventral horn choroid fissures. The supraoptic nuclear field is the primary contributor to the latter group. Scattered neurosecretory neurons are found in association with both pathways, usually in contact with blood vessels supplying the choroid plexuses, or in the telencephalic subependymal stroma. Neurosecretory fibers and terminals occur within the choroid fissures and juxtaventricular neuropil. The neurosecretory terminals in the choroid fissures appear as Herring-body type neurohemal organs; in the neuropil they appear as punctate peri-neuronal desities suggestive of synaptic contacts. Thes morphologic findings are discussed in relation to reports indicating the presence of antidiuretic, vasopressin-like activity in cerebrospinal fluid and choroid plexus extraxts together with ultrastructural evidence supportive of vasopressin-mediated transchoroidal cerebrospinal fluid absorption. These results and those of others indicated the possible involvement of the neurosecretory system in the regulation of brain interstitial-ventricular cerebrospinal fluid dynamics.


Capacity of ribosomal RNA promoters of E. coli to bind RNA polymerase. The rate of in vitro transcription of the rRNA genes of E. coli is more than 20 fold higher than the averaged transcription rate of other genome segments of the same size. This "preferential transcription" of rRNA genes reflects a high efficiency of their promoters in chain initiation. We show that the high initiation rate at rRNA promoters results from a high rate of RNA polymerase binding to these promoters as measured by the formation of heparin-resistant RNA polymerase-DNA complexes. The results indicate that the preferential binding of RNA polymerase to rRNA promoters is mainly due to their large binding capacity rather than to a high rate constant of polymerase binding to a single binding site. The polymerase binding capcity of rRNA promoters was estimated from the number of rRNA chains initiated by heparin-resistant complexes under conditions of template saturation and from the number of rRNA transcription units participating in the binding reaction. At least 30 RNA polymerase molecules were found to be protected from heparin per rRNA transcription unit. The rest of the genome (99.4%; possibly sufficient to encode 4000 nonribosomal RNA species) protects under these conditions 2000 enzyme molecules. These results suggest that a high multiplicity of RNA polymerase binding may be responsible for the high efficiency of rRNA promoters. The validity of this hypothesis is discussed.


Clonal growth and maturation to immunoglobulin secretion in vitro of every growth-inducible B lymphocyte. The frequency of normal murine B lymphocytes initiating growth in diluted suspension cultures in the presence of a B cell mitogen, such as lipopolysaccharide, can be increased approximately 10(4) fold by the addition of 2 X 10(6) normal thymus cells per ml. This increase in the frequency of growing cells by thymus cells can also be observed with X63-AG8 myeloma tumor cells secreting IgG1. Thus thymus cells may not contribute growth-stimulating factors, but may supply growth-supporting factors. Culture medium and plastic dishes can be conditioned by preincubation with thymus cells for a day after which the thymus cells may be omitted from further culture for maximal B cell growth. Irradiation of thymus cell abolishes their growth-enhancing properties. Thymus cells can be syngeneic and allogeneic with the growing B cells. The frequency of growing LPS-reactive, normal B cells in spleen of 6-8 week old C3H/Tif mice was determined by limiting dilution analysis to be one of three splenic B cells. With this limiting dilution analysis, it was also shown that the cloning efficiency of XB3-AG8 myeloma tumor cells in suspension culture in the presence of thymus cells is practically 100%. Analysis of the growth kinetics of single clones of LPS-reactive, normal B cells shown that these B cells divide every 18 hr. Within the first 126 hr of growth, every B cell in the clone divides, and every dividing B cell in this clone secretes sufficient immonoglobulin to form a hemolytic plaque. The conditions of in vitro suspension cultures of murine B lymphocytes are therefore perfect to the extent that every B cell capable of growth will grow as a single clone.


Evolutionary drift of the argF and argl genes. Coding for isoenzyme forms of ornithine transcarbamylase in E. coli K12. Considerable genetic drift has occurred during the evolution of the two genes, argF and argl, which individually code for isoenzyme forms of ornithine transcarbamylase in E. coli K12. The use of the experimental protocol described in this work established that between 25-40% of the base pairs in the genes argF and argl have changed since they diverged from a hypothetical ancestral gene. The extent of divergence of the genes was determined by mRNA-DNA hybridization utilizing arginine transducing DNA as a hybridization probe and mRNA prepared in vivo from appropriate bacterial strains and with mRNA synthesized in vitro using template DNA isolated from the specialized transducing phages gammacI857dargl and phi80dargF.


Sequential biochemical and morphological events during assembly of the fertilization membrane of the sea urchin. The fertilization membrane of Strongylocentrotus purpuratus undergoes changes in morphology, solubility, and permeability during the process of hardening. As the fertilization membrane elevates from the egg surface, it retains casts of the tips of the microvillous processes of the plasma membrane. The dome-shaped microvillar casts become angular at the same time that the fertilization membrane becomes resistant to solubilization in mercaptan solutions. 2-4 min after this morphological and chemical transition, the fertilization membrane becomes impermeable to the lectin conconavalin A, as monitored by binding of 125I- or fluorescein-labeled concanavalin A. Glycine ethyl ester inhibits the changes in morphology, solubility, and permeability, whereas sodium sulfite inhibits only the permeability block and resistance to solubilization by mercaptans. Parthenogenetic activation with the divalent ionophore, A23187, elicits fertilization membrane elevation more rapidly than does activation by fertilization; however, the morphological and permeability changes characteristic of hardening proceed more slowly. Elevation and hardening of the fertilization membrane thus appear to be discrete, multiple-step assembly processes that occur in fixed sequence, with kinetics that are affected by the mechanism of cortical granule exocytosis.


The interaction of an anti-tumour platinum complex with DNA. The interaction of the anti-tumour active cis platinum (II) complexes with DNA has been investigated using dichloro(ethylenediamine)platinum(II) and E. coli DNA. Equilibrium dialysis studies indicate that Pt(en)Cl2 binds reversibly to DNA to a saturation value of 0.57 Pt: P, which is consistent with the platinum being bound both monofunctionally and bifunctionally. Pt(en)Cl2 inhibits the intercalation of 9-aminoacridine (9AA) by cross-linking the bases of the double helix, but at no stage does all the bound platinum cross-link. It is suggested that this inhibition of intercalation is due to intrastrand cross-linking.


Regional myocardial perfusion in patients with atherosclerotic coronary artery disease, at rest and during angina pectoris induced by tachycardia. We studied regional myocardial perfusion by scintigraphic computer-assisted analysis of initial distribution, washout rates, and residual activity of 133Xe injected into the left coronary artery of four patients with normal arteriograms and 14 patients with coronary stenosis. At rest, residual activity in poststenotic regions was always greater than in control regions, but initial washout rates were not slower. During angina, following xenon injections, the amount of indicator distributed to the poststenotic regions was markedly reduced; the increase of the initial washout rates was smaller than in control regions relative to rest, and residual activity was higher. Initial washout rates did not differ as much as from those of normal myocardium because in severe ischemia too little indicator is deposited initially in these regions to produce a change of any magnitude. Indeed, when angina was induced immediately after the xenon injection, poststenotic washout rates became much slower during angina than at rest, a finding that implicates functional factors in impairing poststenotic myocardial perfusion during angina.


Evaluation of the CO2 rebreathing cardiac output method in seriously ill patients. The CO2 rebreathing cardiac output method is a totally noninvasive Fick procedure needing validation in various disease states to become clinically applicable. Simultaneous measurements of cardiac output by CO2 rebreathing and dye-dilution or direct Fick techniques were performed in 53 patients. In nine patients with pulmonary disease rebreathing cardiac output averaged 4.85 L/min compared to 5.18 L/min by dye-dilution or Fick (r = 0.16). In 14 instances of acute myocardial infarction cardiac output was 5.53 L/min by rebreathing and 5.87 L/min by dye-dilution (r = 0.95), while in nine shock cases it averaged 3.98 L/min by dye-dilution or Fick and 3.75 L/min by CO2 rebreathing (r = 0.94). In five heart failure cases with mitral insufficiency, which may distort dye durves, correlation between standard and rebreathing methods was r = 0.09, but in 16 cases without mitral regurgitation, r = 0.89. Acute interventions in ten patients increased dye-dilution cardiac output by 0.92 L/min and rebreathing outputs by 0.60 L/min (r = 0.87). The data suggest that the CO2 rebreathing cardiac output method may be useful in the CCU-MICU setting.


Multiclinic controlled trial of bethanidine and guanethidine in severe hypertension. One hundred and eight patients with initial diastolic blood pressure in the range of 100-124 mm Hg while taking hydrochlorothiazide were assigned randomly and double-blind to hydrochlorothiazide plus either bethanidine or guanethidine. The average reduction of the fifth and sixth months' diastolic blood pressure was 18.4 mm Hg for guanethidine and 13.6 mm Hg for bethanidine (P less than 0.01). The distribution of the individual values was such that 68.8% of guanethidine treated patients achieved a diastolic level below 90 mm Hg, compared to only 45.5% of the bethanidine treated group (P less than 0.025). The degree of orthostatic fall in blood pressure was greater with bethanidine than with guanethidine (P less than 0.05). The diurnal variation of blood pressure was slightly greater with bethanidine than with guanethidine. The results significantly favor guanethidine. This  study failed to demonstrate that the shorter action of bethanidine confers significantly better control of blood pressure than the longer action of guanethidine.


Aseptic necrosis of the femoral head. Treatment with bone grafts of doubtful value. A heterogeneous group of 17 patients with aseptic necrosis of the femoral head had treatment of 23 hips by bone grafting. Phemister-type tibial grafts were used in 20 and iliac grafts in 3. All hips were evaluated before and after grafting according to roentgenographic stage of disease and the Hospital for Special Surgery Rating System. We found no exact correlation between roentgenographic stage and clinical manifestations. It is concluded that bone grafting treatment of this condition is warranted in only the very earliest stage of the disease when roentgenographic examination shows that normal femoral head contour is preserved and there are only subtle mottled densities scattered throughout the anterosuperior region. In this series, no benefit was derived from bone grafting in later stages of the disease.


Serratia marcescens arthritis in heroin addicts. Serratia appears as a pathogen of increasing frequency and clinical significance in bone and joint infections in heroin users. This is the fifth case report of septic arthritis due to Serratia marcescens in intravenous heroin users. The clinical and laboratory features were not different from other acute pyogenic arthritides. Signs of infection were obscure even in the presence of debilitating disease. Although Pseudomonas and Staphylococcus are more common organism in bone and joint infections of heroin users, Serratia should be considered as a possible pathogen in such patients. In the present case, immediate open drainage followed by systemic Gentamicin treatment gave rapid relief of pain and restoration of full range of motion of the joint.


Treatment of steroid induced osteocartilagenous separation of the distal femur. A case report. A 20-year-old Negro man, with a history of febrile disease, was diagnosed to have noncaseating granulomatous inflammatory disease. He was placed on Prednisone therapy for several years. Subsequently, he was found to have osteocartilagenous separation of the lateral condyle of the right femur. This was preceded by symptoms of recurrent pain and swelling of the knee. The surgical treatment employed was anatomic repositioning of the osteocartilagenous fragment with an interposed corticocancellous graft. The postoperative management was non weight-bearing for a 6 months' period and a cast brace. The patient was symptom free at 20 months' follow-up.


Meralgia paresthetica. An anatomical interpretation. One hundred and ten lateral femoral cutaneous nerves of the thigh were dissected in 90 adult and 20 fetal cadavers. In 51 per cent of adult cases a significant enlargement or pseudoganglion was present in the area of the nerve where it passed under the guinal ligament to turn sharply downward onto the thigh. The histological examination of these pseudoganglions showed a thickened epineurium, perineurium and endoneurium. No pseudoganglions and no histological abnormalities were found in the fetal nerves. It is proposed that erect human posture, combined with the particular course of the lateral femoral cutaneous nerve are the causes of tension, mechanical friction, and irritation of the nerve, and contribute to the development of the pseudoganglion and play a role in the pathogenesis of idiopathic meralgia paresthetica.


Reduction in plasma norepinephrine during fenfluramine treatment. Plasma norepinephrine levels were reduced to 30% of placebo levels during the administration of fenfluramine, 120 mg/day, for 10 or more days. There was also a significant reduction in plasma dopamine beta-hydroxylase activity during fenfluramine treatment, suggesting that chronic administration may induce antiadrenergic effects which may be useful in the treatment of disorders affected by circulating catecholamines.


Electrically stimulated bone growth in animals and man. Review of the literature. The literature on the electrical stimulation of bone growth and fracture healing has been increasing exponentially in recent years. About 95 per cent are positive reports despite an extraordinarily wide selection of experimental techniques and models. Fourteen research groups report that electrical currents stimulated fracture healing with few if any complications in a total of 595 patients. The mechanisms of action and ideal technique for applying stimulation has yet to be determined.


Metolazone and spironolactone in cirrhosis and the nephrotic syndrome. Eighteen patients with hepatic cirrhosis or nephrotic syndrome and having edema and/or ascites were treated during successive periods with metolazone 5 to 40 mg/day, spironolactone 100 mg/day, and with both diuretics concurrently. Metolazone alone produced a marked diuresis, natriuresis, and weight loss in 8 patients. Spironolactone alone had little effect, but the addition of metolazone renewed diuresis and natriuresis and resulted in additional substantial weight losses in all patients responsive to metolazone alone. Concurrent spironolactone and metolazone also induced moderate diuretic effects in some patients who failed to respond to either drug alone. The drugs were well tolerated; the administration of spironolactone with metolazone prevented decreases in serum potassium, which had occurred during treatment with metolazone alone.


Guanabenz in essential hypertension.

Guanabenz in essential hypertension. Fifty-five patients with mild to moderately severe essential hypertension were treated with guanabenz (2, 6-dichlorobenzylidene aminoguanidine acetate) in doses from 4 to 16 mg twice daily in a randomized, placebo-controlled study. The patients treated with placebo in the initial phase of the study were subsequently treated with guanabenz. The mean arterial pressure in the guanabenz group decreased from 130.6 to 107.6; that in the placebo group decreased from 129.6 to 126.6 standing and from 126.6 tp 109.9 and 128.8 to 120.5, respectively, supine. The principle adverse effects included sedation, dry mouth, weakness, and tiredness. Of the guanabenz-treated patients 84% had sustained decrease in supine diastolic blood pressure of 10 mm Hg or more, whereas in the placebo-treated patients only 32% had such a response. There was no significant orthostatic hypotension. Guanabenz thus appears to be an effective antihypertensive drug in patients with mild to moderately severe hypertension.


Absorption of rifampin from various preparations and pharmaceutic forms. The absorption of 9 rifampin preparations was measured in two seaparate crossover studies on 10 healthy female volunteers. In the first set of experiments, one of the three capsule preparations was absorbed more slowly than the other ones. The absorption of two syrup preparations was nearly twice that of the best capsule preparation. In the second set, the absorption of the commercial tablets appeared to be somewhat faster than that of the reference capsule preparation. One tablet dissolved slowly in vitro and yielded very low serum concentrations. On the basis of these results the equiactive dose of rifampin in syrup form may be 25% to 50% lower than that of the other forms.


The classic. Arthrodesing operations on the feet: Edwin W. Ryerson,M. D. The concept of stabilizing paralytic or otherwise deformed joints by surgical fusion started with Eduard Albert of Vienna in the latter 19th century. His classic paper on arthrodesis of the knee appeared in its first English translation in this journal in 1946 (No. 34) (see Classics of Orthopaedics, Phila. J. B. Lippincott Co., 1976). However before his success in the knee in 1882, he had attempted fusion of the tarsus. Immediately following Albert's publication, attempts at fusion of the tarsus were published by Golding-Bird in England, Ogston of Scotland, and many others up to Gwilyn G. Davis of Philadelphia in 1913 whose subtalar arthrodesis became the popular procedure for a decade and seriously competed with Royal Whitman's astragelectomy. In 1923 Edwin W. Ryerson of Chicago (1872-1961) designed a surgical procedure for which he used the term triple arthrodesis. This operation, with minor modifications, became standard and except for the basic concept of Albert, remains among the classics in this subject. It was a relatively frequent procedure when polio was a world-wide orthopedic problem, but has continued a valuable part of the armamentarium of foot surgery whenever trauma or other cause leaves a painful deformed foot.


Chymopapain--past and present, future?

Chymopapain--past and present, future? Clinical experience with Chymopapain casts doubts upon the reported results of the limited double blind studies done mainly by surgeons and their residents, relatively inexperienced in the disc injection technique. Chymopapain is a useful drug when used properly for lumbar disc disease and has the potential of eliminating three-fourths of lumbar disc surgery. Chymopapain should have been released by the FDA on a Phase IV basis. In the mean-time the drug is available in Canada and England but not in the U.S.


Intermittent mandatory ventilation in the neonate. Intermittent Mandatory Ventilation (IMV) provides an alternative method of support to assisted and controlled mechanical ventilation in neonates with acute respiratory failure. Specific advantages include the use of only the level of mechanical support which is required on an individual basis. Because intrapleural pressure ventilation (IPPV), venous return and cardiac output are maintained at more nearly normal levels, with or without positive end-expiratory pressure (PEEP). Duration of mechanical ventilatory support is reduced, weaning enhanced, and the incidence of pulmonary barotrauma decreased with IMV. Physiological homeostasis in terms of acid-base changes is more readily maintained as a result of precise regulation of alveolar ventilation. Newer concepts in mechanical support, such as "reversed" inspiratory:expiratory ratio (I:E ratio) ventilation, are possible with IMV because of the lower ventilator rates employed compared to IPPV.


Cutaneous candidiasis: treatment with miconazole nitrate. In a well-controlled, double-blind, randomized study, 30 patients with cutaneous candidiasis were treated with a 2% miconazole nitrate lotion or its placebo control. By the 14th day, 13 of the 15 patients [87%] treated with miconazole nitrate achieved clinical and mycologic cures. Only a single patient treated with the placebo lotion would be classified as a therapeutic cure. In a second portion of the study those patients judged to be therapeutic failures were treated with the lotion containing 2% miconazole nitrate. By combining the results of both portions of the study we find that miconazole nitrate lotion produced both a clinical and mycologic cure in all patients receiving the active lotion. The miconazole lotion formula was well tolerated by all patients and no side effects were noted. The fact that miconazole nitrate acts rapidly in relieving symptoms, is well tolerated, and is highly effective against dermatophytes, yeasts and gram-positive bacteria, makes it a welcome addition to available topical therapy of skin infections.


Candidiasis seminar.

Candidiasis seminar. This is a review of candidiasis from its historical, epidemiologic aspect through the laboratory diagnoses, precipitating factors and the more common diseases caused by Candida. Therapeutic modalities available are listed and specific guidelines for therapy are demonstrated.


Widespread cutaneous herpes simplex type II infection. The case of an adult who survived a widespread culture-proven cutaneous herpes simplex type II infection is presented. The authors believe it is the first such case. The virus was demonstrated by its cytopathic effect in Vero cell culture. Large, swollen syncytial cells and holes in the cell sheet, typical of the changes produced by the herpes simplex virus type II, were noted. Photodynamic inactivation therapy with acriflavine dye and fluorescent light was employed.


Plasma levels of lidocaine following nebulized aerosol administration. The venous (plasma) levels of lidocaine were measured in five subjects following nebulization of a 280-mg dose via intermittent positive pressure breathing (IPPB) and nebulization of a 400-mg dose via ultrasound. Even though a lower dose of lidocaine was given by IPPB, this system of delivery produced higher plasma concentrations of the drug than ultrasound and also was more effective in eliminating the gag reflex. In the entire study, plasma concentrations of lidocaine did not exceed 1.1 mug/ml, which is far below toxic levels. Aerosolized lidocaine, administered by IPPB, is a safe and effective topical anesthetic agent which may be useful for instrumentation of the upper airway.


Treatment of chronic salmonella carriers. Study with 40 cases of S. typhi, 19 cases of S. paratyphi b and 28 cases of S. enteritidis strains. The present stage of our studies suggests that, provided a highly effective combined therapy is, and can be, carried out correctly, all excreters can be cured of their chronic carrier state by chemotherapy within 8-12 weeks. Although we cannot recommend a universal therapeutic regimen for all patients, a highly effective 'basic therapy' (RMP+TSP) is available for the majority of the cases, needing occasional modification, depending on the specific requirements of the individual patient as shown by the result of the serum activity determination. This method saves the patient from toxic inconveniences caused by inadequate treatment, it shortens the treatment time and makes cholecystectomy superfluous - unless it is considered necessary out of a different indication in which case it should certainly be done. We cannot share the often expressed view that Salmonella enteritidis excreters cannot be cured, a view which is found even in the most recent manuals. The same applies to the view that therapy is not necessary because it would delay cure. It is indispensable to establish a close cooperation between the public health authorities and the private physician, and we therefore wish to sincerely thank all colleagues and Public Health Officers for their collaboration.


[Choice of transplant in cranial defects. Analysis of 73 cases]. In many of the increasing number of primary or secondary calotte defects resulting from traumatic cranial injuries, plastic reconstruction of the cranium is required. Between 1965 and 1975 in the Surgical Department of Lucerne's Canton Hospital 73 posttraumatic defects of the cranium were repaired. In 69 cases autoplastic transplants and in 4 cases alloplastic transplants were used. The calotte plate (Elliot and Scott), pieces of pelvic bone (Converse), rib (Longacre and De Stefano), as well as costal cartilage (Peer) provided autoplastic material. Alloplasty for the cranium was carried out only in selected cases where special indications were present. Our results in the selection of autologous transplants for the reconstruction of secondary cranial defects is discussed and the surgical procedure is presented.


[Treatment of flail chest from the general surgical view. Experience with 20 cases]. Flail chest is a serious complication of multiple fractures of the ribs that needs immediate management in order to avoid its disastrous respiratory and circulatory consequences. The authors present their experience with 20 cases of flail chest injury and discuss especially some practical aspects of its treatment. Skeletal traction was used frequently to stabilize the chest wall, while intermittent positive pressure respiration was reserved only for patients with uncompensated respiratory disturbances. The authors urge that a well-organized surgical unit should be able to provide appropriate management in such cases, since most of these patients have multiple injuries that need general surgical together with more specialized treatment.


Pimozide versus fluphenazine in ambulatory schizophrenics: A 12-month comparison study. In this study, chronic schizophrenic outpatients who had been maintained on various neuroleptics for an average of about 4 years had their previous medications (approximately equivalent to 695 mg of chlorpromazine per day) changed abruptly to either pimozide or fluphenazine given in single daily oral doses on a double-blind basis for a period of 52 weeks. Average daily doses were pimozide 9.6 mg and fluphenazine 12.5 mg. Measurements of the therapeutic effects of the two drugs were made immediately prior to starting the study, at the end of the 2nd and 4th weeks, and thereafter every 4th week to the end of the study. Three psychometric scales were used for evaluation: Brief Psychiatric Rating Scale (BPRS); Evaluation of Social Functioning (ESFR); and Clinical Global Impressions (CGI). In addition, patients participated in a Social Adjustment Inventory (SAI) evaluation. Statistical analysis with the use of several statistical techniques for between- and within-drug group comparisons revealed that pimozide and fluphenazine were equally effective in maintaining control of symptomatology of chronic schizophrenics at a level commensurate with or better than that provided by their previous medication. Side effects were characteristic of marketed neuroleptics, similar in severity and occurrence between study-drug groups, mainly extrapyramidal symptoms, and readily controlled with antiparkinsonian medication. Pimozide, slightly more potent than fluphenazine, proved to be equally effective for the long-term management of chronic schizophrenic patients.


[Is cytarabine an effective drug against herpes zoster? (author's transl)]. Although herpes zoster is a viral infection with spontaneous healing, the skin eruption is painful and there may be serious complications. Cytarabine was administered to 91 patients with the disease. It was given by quick infusion or intravenously twice daily for five days. An open, uncontrolled study of 61 patients suggested that the drug was beneficial without serious side effects. But in a double-blind randomised study of 30 patients there was no significant effect on the course of the disease. The mean healing time for skin blisters in segmental uncomplicated zoster was 8.4 days with cytarabine and 6.5 days with a placebo.


Anthelmintics.

Anthelmintics. This article describes the drugs used in helminthic infections and their therapeutic indications, mode of action, toxicity and other details of each of the recommended drugs, and discusses the nature and treatment of infection by helminths important in human medicine. Infestation due to the roundworms Enterobius vermicularis, Ascaris lumbricoides and the hookworms, Ancylostoma duodenale and Necator americanus can all be treated effectively with pyrantel pamoate. For Enterobius vermicularis, however, a newer drug, mebendazole, is equally as effective. The advantage of these drugs in the indicated circumstances is that they can be administered in a single dose. Unfortunately, pyrantel pamoate is not a panacea and in the case of Necator it is not as effective as in the other roundworms. In that situation the use of tetrachlorethylene is preferable. For treatment of Strongyloides stercoralis, and important human parasite, because it can become disseminated and lead to fatal infections in immunoincompetent hosts, the only effective drug is thiabendazole. In treatment of Trichuris trichiura infection, mebendazole, administered over a period of 3 days, is the most effective available drug. For the roundworms inhabiting tissues--either as aberrant infections of man or as the normal part of their life cycle in man--therapy tends to be largely non-specific. For example, in visceral larva migrans, caused by the dog roundworm Toxocara canis, only palliative therapy with systemic anti-inflammatory agents and corticosteroids may be helpful. Cutaneous larva migrans, caused by the dog hookworms Ancylostoma brasiliensis and Ancylostoma caninum, is also treated primarily with symptomatic measures, but there is a suggestion that thiabendazole may kill the larvae and thus be effective. Trichinella spiralis may cause severe, even fatal infections in man, but only symptomatic therapy can be offered. Therapy for filarial infections is regrettably complicated and not completely effective. Diethylcarbamazine remains the best available drug, but in some of these infections local surgical excision may also be used. It is important to bear in mind that release of antigens from dying or dead worms may cause systemic inflammatory and allergic reactions that may require therapy with corticosteroids. Therapy for Cestodes is achieved most effectively with niclosamide, but the antimicrobial agent paromomycin has also been effective. For the aberrant cestode infections of man, such as echinococcal cysts or Taenia solium cycticerci, treatment is surgical if the affected areas are accessible. Treatment of schistosomal infections is quite toxic and, therefore, it is mandatory to determine viability of the worms before recommending therapy. If therapy is required, then Schistosoma mansoni infections are treated with stibophen and S. japonicum with antimony potassium tartrate, taking care in both of these instances to watch for the early signs of antimony toxicity; therapy of S. haematobium infections is based on administration of niridazole...


Digitalis in pulmonary heart disease (cor pulmonale). The use of digitalis in pulmonary heart disease has been a topic of great interest for a number of years. The physician's decision to use or not to use digitalis in pulmonary disease has often been an emotional rather than a reasoned one. The diagnostic difficulties from a clinical point of view in separation of pulmonary from cardiac symptoms and findings have also been confusing. The fact that small doses of digitalis may have an inotropic effect on the cardiac muscle has been a difficult concept for many physicians to adopt. On the other hand, the larger doses of digitalis that are often necessary to control the ventricular response in supraventricular arrhythmias sometimes gives rise to confusion. We shall attempt to review the subject in detail and examine indications, contraindications, toxicity, dosage, assessment of benefit, and role of digitalis serum levels in patient management.


Inhibition of in vivo pituitary release of luteinizing hormone in lactating rats by exogenous prolactin. The mechanism whereby the reciprocal relationship between the plasma levels of prolactin and HL is maintained in lactating rats under different degrees of suckling stimulus has been investigated in the present study. Plasma levels of luteinizing hormone (LH) in lactating rats suckling two pups could be reduced significantly by injecting prolactin (PRL). This reduction was also evident in ovariectomized and ovariectomized-adrenalectomized lactating rats, thus excluding mediation of the inhibitory effect by steroids from end-organs. The in vivo response of the pituitary to exogenous LHRH was lower in rats suckling eight pups than those suckling two pups. Prolactin administered prior to LHRH caused an inhibition of the response of the pituitary to exogenous LHRH in rats suckling two pups. These data provide evidence for the hypothesis that in the lactating rat, in the presence of minimal suckling stimulus, the causative factor for reducing serum LH levels is prolactin, which acts by altering the pituitary responsiveness to LHRH.


Stimulation of proinsulin biosynthesis by purine-ribonucleosides and D-ribose. Inosine and guanosine were potent stimuli of proinsulin biosynthesis ([3H]leucine incorporation) in isolated pancreatic islets of the rat. The effect was nearly abolished by formycin B, an inhibitor of purine nucleoside phosphorylase, but not by D-mannoheptulose. The corresponding bases had no effect on the rate of proinsulin biosynthesis. D-ribose enhance proinsulin biosynthesis at low concentrations )0.3-0.6mM) but concentrations above 5 mM were ineffective. The effect of all three compounds was highly specific for proinsulin biosynthesis, since incorporation of [3H]leucine into other islet proteins was not significantly stimulated. The data strongly indicate that metabolic signals regulate modulation of proinsulin biosynthesis in the beta cells.


A general survey of tumours in the horse. Published figures from veterinary schools indicate that tumours may account for about 1 to 3 per cent of surgical cases. In this paper an account is given of clinical and pathological aspects of the tumours that are most likely to be encountered in practice, either as visibly or palpably detectable masses of tissue, or as internal tumours responsible for clinical signs. Amongst the common and important tumours dealt with, the most prominent are "equine sarcoid", squamous-cell carcinoma (of eye region and of glans penis), granulosa-cell tumours and melanomas. Well over a hundred references provide entry to the relevant literature. The need for further pathological and clinical studies of equine tumours is stressed.


Coughing in horses--an historical aspect. A brief historical review is given of the incidence and types of respiratory disease that occurred in Britain in the 18th and 19th centuries. The significance of poor stabling and overcrowding in the causation and spread of coughing is emphasised and its dramatic reduction by simple methods of hygiene and ventilation.


Inhibition of translation in eukaryotic systems by harringtonine. The Cephalotaxus alkaloids harringtonine, homoharringtonine and isoharringtonine inhibit protein synthesis in eukaryotic cells. The alkaloids do not inhibit, in model systems, any of the steps of the initiation process but block poly(U)-directed polyphenylalanine synthesis as well as peptide bond formation in the fragment reaction assay, the sparsomycin-induced binding of (C)U-A-C-C-A-[3H]Leu-Ac, and the enzymic and the non-enzymic binding of Phe-tRNA to ribosomes. These results suggest that the Cephalotaxus alkaloids inhibit the elongation phase of translation by preventing substrate binding to the acceptor site on the 60-S ribosome subunit and therefore block aminoacyl-tRNA binding and peptide bond formation. However, the Cephalotaxus alkaloids do not inhibit polypeptide synthesis and peptidyl[3H]puromycin formation in polysomes. Furthermore, these alkaloids strongly inhibit [14C]trichlodermin binding to free ribosomes but hardly affect the interaction of the antibiotic with yeast polysomot interact with polysomes and therefore only inhibit cycles of elongation. This explains the polysome run off that has been observed by some workers in the presence of harringtonine.


Properties of the penicillin-binding proteins of Escherichia coli K12,. Benzyl[14C]penicillin binds to six proteins with molecular weights between 40000 and 91000 in the inner membrane of Escherichia coli. Two additional binding proteins with molecular weights of 29000 and 32000 were sometimes detected. All proteins were accessible to benzyl[14C]penicillin in whole cells. Proteins 5 and 6 released bound benzyl[14C]penicillin with half times of 5 and 19 min at 30 degrees C but the other binding proteins showed less than 50% release during a 60-min period at 30 degrees C. The rate of release of bound penicillin from some of the proteins was greatly stimulated by 2-mercaptoethanol and neutral hydroxylamine. Release of benzyl[14C]penicillin did not occur if the binding proteins were denatured in anionic detergent and so was probably enzymic. No additional binding proteins were detected with two [14C]cephalosporins. These beta-lactams bound to either all or some of those proteins to which benzyl[14C]penicillin bound. No binding proteins have been detected in the outer membrane of E coli with any beta-[14C]lactam. The binding of a range of unlabelled penicillins and cephalosporins were studied by measuring their competition for the binding of benzyl[14C]penicillin to the six penicillin-binding proteins. These results, together with those obtained by direct binding experiments with beta-[14C]lactams, showed that penicillins bind to all six proteins but that at least some cephalosporins fail to bind, or bind very slowly, to proteins 2, 5 and 6, although they bind to the other proteins. Since these cephalosporins inhibited cell division and caused cell lysis at concentrations where we could detect no binding to proteins 2, 5 and 6, we believe that these latter proteins are not the target at which beta-lactams bind to elicit the above physiological responses. The binding properties of proteins 1, 3, and 4 correlate reasonably well with those expected for the above killing targets.


The primary structure of yeast alcohol dehydrogenase. Eight different types of peptide mixtures from [14C]carboxymethylated yeast alcohol dehydrogenase were obtained using trypsin with or without prior maleylation of the substrate, chymotrypsin, pepsin, microbial proteases or CNBr. Each mixture was fractionated by exclusion chromatography and peptides were further purified on paper. From results of analyses of all fragments it seems possible to to deduce a primary structure of 347 unique residues in three segments. Together, the segments can account for the whole protein monomer with the exception of a small connecting region. Many unfavourable structures complicated the determination and made single sequence conclusions tentative, but known data are consistent and for most segments of the monomer results are abundant. Several microheterogeneities in the protein are indicated and one apparent amino acid exchange is characterized, suggesting that different types of subunits occur. This may probably be correlated with genetic polymorphism in yeast. Multiple desamidations are also characterized and a few of these affect particularly labile structures. Many residues are unevenly distributed and unexpected patterns are shown. Elements of repetitive sequences occur, reducing the uniqueness of structures. Hydrophobic segments are found, and the uncharacterized region is, at least in some subunits, in a core-like tryptic segment. These and other aspects of the structure may explain some properties of the monomer, and form the background for evolutionary, structural and functional correlations with related enzymes.


Differences between alcohol dehydrogenases. Structural properties and evolutionary aspects. Comparisons of the primary structures of yeast and horse liver alcohol dehydrogenases reveal that the enzymes are homologous but distantly related. The overall positional identity is 25% between common regions, and several deletions/insertions occur in either enzyme, the longest apparently corresponding to 21 residues, showing that the different subunit sizes are largely explained by internal differences. Variabilities in the structural similarities can be coupled with functional requirements but not directly with whole domains in the previously known tertiary structure of the horse protein. The two most similar regions of the enzymes affect active-site segments and the two most dissimilar regions seem to affect a loop structure without known function, and a segment participating in subunit interactions. The dissimilarities may probably be correlated with changes in zinc-binding properties and quaternary structures. The extra region corresponding to the large internal chain-length difference shows an apparent coincidence in sequence to a following segment of the horse enzyme, and additional elements of internal coincidences, or superficial similarities with other dehydrogenases, are noticed. These characteristics are not fully distinguishable from chance distributions but in view of the extensive species variations in alcohol dehydrogenases some evolutionary considerations may not be excluded, in which case a model relating all regions of these and associated enzymes to a common ancestor is shown to be compatible with all known observations.


Selective inhibition of tRNATyr transcription by guanosine 3'-diphosphate 5'-diphosphate. Guanosine 3'-diphosphate 5'-diphosphate (ppGpp) selectively reduces the synthesis of su+III tRNA from omega 80 psu+III DNA relative to the synthesis of omega 80 RNA in a system in vitro containing DNA and Escherichia coli RNA polymerase holoenzyme as the sole macromolecular components. The response of su+III tRNA synthesis to increasing salt and to temperature in the presence of ppGpp suggests that the nucleotide may reduce the affinity of the enzyme for su+III promoters. The Ki for the selective inhibition of tRNA synthesis by ppGpp is 4 muM in contrast to the value of 150 muM for the inhibition of rRNA synthesis.


Bacteriophage-T7-induced DNA-priming protein. A novel enzyme involved in DNA replication. The T7gene-4 protein has been purified to near homogeneity using a complementation assay in vitro, and it is designated T7 DNA-priming protein (DNA primase). The purified enzyme enables T7 DNA polymerase to initate DNA synthesis on various circular single-stranded DNA templates by a mechanism which involes the synthesis of a very short RNA primer. The oligoribonucleotide, which is linked to the product DNA via a 3':5'-phosphodiester bond, starts with pppA-C and terminates predominantly with AMP. When only ATP and CPT are precursors, the RNA primer is found to be primarily a tetranucleotide of the sequence pppA-C-C-A. Using oligoribonucleotides in place of ribonucleoside triphosphates as chain initators, T7 DNA-priming protein drastically  increases the efficiency with which T7 DNA polymerase can utilize particular tetranucleotide primers containing A and C residues. T7 DNA-priming protein also enables T7 DNA polymerase to make use of native or nicked duplex T7 DNA as template-primer. This reaction does not require ribonucleoside triphosphates, although their addition enhances DNA synthesis 2--4 fold. The product formed in their absence is covalently attached to the template DNA and is found to contain a few long branches when examined by electron microscopy. In the presence of ribonucleoside triphosphates most of the newly made product arises from imitation of DNA chains de novo. Incubation of three proteins: T7 DNA-priming protein, T7 DNA polymerase, and T7 DNA-binding protein, with ribonucleoside and deoxyribonucleoside triphosphates, and with phiX174DNA as template leads to the generation of 'rolling circle-like' structures as visualized in the electron microscope. Single-stranded regions at the tail-circle junction indicate that initations can occur de novo on the displaced complementary strand. This is consistent with a discontinuous mode of 'lagging' strand synthesis and suggests that the same proteins may also be responsible for fork propagation in vivo.


The functional and fluorescence properties of Escherichia coli RNA polymerase reacted with fluorescamine. 1. Fluorescamine (4-phenylspiro[furan-2,(3)1'-phthalan]-3,3'-dione) reacts rapidly with Escherichia coli RNA polymerase and produces a fluorescent derivative which is inactivated to an extent dependent upon reagent concentration. Excess fluorescamine is rapidly hydrolysed. Reaction is with xi-amino gruops of lysine residues in all subunits as revealed by gel electrophoresis and fluorescence scanning. 2. The extent of inactivation and fluorescence yield are diminished in the presence of added template, a finding which provides evidence for the existence of reactive and essential amino groups which can be at least partially shielded by DNA in the binary complexes. The relative decrease of fluorescence is greatest in the betabeta' subunits. Holoenzyme and core enzyme show essentially the same behavior. 3. The inactivation of activity by fluorescamine is primarily at the level of initiation. Template binding and chain propagation are less affected. 4. The enzyme derivatized by fluorescamine shows an intense fluorescence with a peak at 490 nm and an excitation maximum at 390 nm. The fluorescence lifetime is in the range of 3-8 ns and the emission is highly polarized. In reactions carried out at high ionic strength the fluorescence yield is approximately double that at low ionic strength and insensitive to the presence of template. 5. Energy transfer is observed between the derivatized enzyme as donor and ethidium bromide as acceptor in the presence of template to which both the enzyme and intercalating dye are bound. The transfer efficiency is a function of the relative concentrations and of the conditions of reaction with fluorescamine. An average transfer distance of approx. 4-5 nm has been calculated suggesting a close proximity between bound polymerase and helical regions of the template.


The amino-acid sequence of the dihydrofolate reductase of a trimethoprim-resistant strain of Escherichia coli. The determination of the amino acid sequence of the dihydrofolate reductase from Escherichia coli RT500 is described. The sequence, comprising 159 residues, has been derived from automatic sequencing of the intact protein in conjunction with manual sequencing of lysine-blocked tryptic peptides, Staphylococcus aureus protease peptides, and alpha-lytic protease peptides. Comparison of the sequence with that of the dihydrofolate reductase from a methotrexate-resistant strain of E. coli (MB1428) shows that 145 of the residues are identical. The distribution of the differences along the length of the molecule is discussed.


Transcription of double-stranded RNA by Escherichia coli DNA-dependent RNA polymerase. Double-stranded RNA of some virus genomes can be used as template for the DNA-dependent RNA polymerase purified from Escherichia coli. The RNA synthesis requires all four nucleoside triphosphates and manganese ions and is dependent on the presence of sigma subunit. The reaction is inhibited by rifampicin, streptolydigin and ethidium bromide, but not by DNase and actinomycin D which does not bind to double-stranded RNA. The template activity of double-stranded RNA from various viruses is different in each case. The order of template efficiency is Penicillum chrysogenum virus greater than cytoplasmic polyhedrosis virus greater than rice dwarf virus greater than reovirus. The product obtained using cytoplasmic polyhedrosis virus double-stranded RNA as template is single-stranded and hybridizes specifically to the denatured template RNA. One of the major 5'-starting nucleotide sequences of the product RNA is pppA-A-Y--. These results indicate that transcription in vitro of double-stranded RNA by E. Coli RNA polymerase is initiated at specific sites on the template.


Processing by ribonuclease II of the tRNATyr precursor of Escherichia coli synthesized in vitro. The tRNATyr precursor molecule, synthesized from phi 80 psu3+ DNA (containing a single tRNA gene) by DNA-dependent RNA polymerase and q factor, was about 205 nucleotides long. The main product of its digestion with a ribonuclease tii preparation from Escherichia coli showed the same electrophoretic mobility as tRNAtyr precursor isolated in vivo and was found to be identical to it when analysed using fingerprint techniques. This intermediate precursor synthesized in vitro was converted further by processing with ribonuclease P into an RNA identical size to mature tRNATyr. It was concluded that the initiation of transcription of the tRNATyr gene in vitro occurs at the same site as that of transcription in vivo and a termination occurs at about 80 nucleotides beyond the CCA end of tRNATyr.


Simple isolation method and assay for T4 DNA ligase and characterization of the purified enzyme. A method for the isolation of T4-amber-N82-induced DNA ligase is described which results in a nearly homogeneous enzyme preparation after two column chromatographic steps. The enzyme is detected during the purification by its ability to form a stable acid-precipitable enzyme-adenylate complex. Some properties of the assay, such as the effect of salt, temperature and incubation time, are presented. The isolated enzyme and its adenylate complex are characterized by acrylamide gel electrophoresis under native and denaturing conditions, as well as by isoelectric focusing. The purified enzyme exhibits a molecular weight of approximatel 60000. Isoelectric focusing yields at least 5 protein components, which are able to form an enzyme-adenylate complex. The main activity possesses a p1 of 6. The enzyme preparation is capable of repairing T5+ DNA known to contain about 4 or 5 single-strand breaks, to circularize lambda DNA and to join Hind111 and EcoR1 fragments.


Non-equivalence of the sites of yeast phenylalanyl-tRNA synthetase during catalysis. Yeast phenylalanyl-tRNA synthetase, an enzyme with an alpha2beta2 structure, has two active sites for phenylalanine, tRNAphe, phenylalanyladenylate and phenylalanyl-tRNAphe. Determination of phenylalanine binding properties to the free enzyme by equilibrium dialysis shows that only one mole of amino acid binds per mole of enzyme, i.e. absolute negative cooperativity. Binding of the amino acid in the presence of tRNA or of ATP and PPi unmasks the second phenylalanine binding site. The difference between the affinities at the tight and loose binding sites under such conditions is about 10--15. Titration of phenylalanyladenylate sites by the burst of ATP consumption shows the formation of a (enzyme-phenylalanyladenylate)2 complex in the presence of pyrophosphatase; however, the two sites differ widely in their affinity as shown by dialysis experiments. Measurements of hydrolysis rates of enzyme-bound phenylalanyladenylate suggests that when only the high-affinity adenylate site is occupied, the other protomer can still bind phenylalanine and ATP (in the presence of phenylalanine). Two moles of Phe-tRNAphe bind to the enzyme with a very high affinity (Kd less than 48 nM). The presence of millimolar concentrations of ATP, phenylalanine and pyrophosphate triggers negative cooperativity and under these conditions only one mole of Phe-tRNAphe is bound per mole of enzyme with a Kd value of 0.15 muM. The present results give support to interprotomer catalytic cooperativity in the mechanism of action of yeast phenylalanyl-tRNA synthetase.


The importance of adrenergic receptors in disorders of micturition. A review is presented of the adrenoreceptors in the lower urinary tract, and their relationship to disorders of micturition. It is suggested that they may be of importance of neurogenic bladder disease, bladder neck obstruction, benign enlargement of the prostate, and certain drug and hormonal effects on the lower urinary tract, and that modification of their activity can be of benefit in the management of these conditions.


A method for counting ectopic beats by computer analysis of R-R intervals. A method is described for detecting and counting ectopic beats using a small computer. The occurrence of an ectopic beat is identified by the relative duration of adjacent pulse intervals as demonstrated by simple arithmetical procedures. The method was validated by manual counting  of records from 18 patients. 89% of 1474 ectopics were correctly identified. There were few false positive identifications 42. Although the method is unsuitable for the diagnosis of arrhythmias it is likely to be of value in epidemiological studies of such disturbances.


Plasma levels and half lives of thioridazine and some of its metabolites. I. High doses in young acute schizophrenics. Plasma levels of thioridazine and four of its metabolites were determined in a series of fifteen young acute schizophrenics; Consistent individually different metabolic patterns were detected in a group of patients who had the same value for the sum of thioridazine plus metabolites. The apparent volume of distribution and half life were calculated. Clinical evolution tended to correlate best with the level of mesoridazine.


Antiarrhythmic and electrocardiographic effects of single oral doses of disopyramide. Ten patients with various heart diseases and ventricular arrhythmia received a single oral dose of disopyramide (DE) 200 mg. The ECG was recorded continuously for about 50 h from 2-4 h before drug administration. A statistically significant reduction in the number of ventricular ectopic beats (VEBs) was seen 1.0-3.5 h after drug intake; the average number of VEBs per 30 min decreased from 317 during the control period to 92 by 1.0-3.5 h after treatment and if one patient who did not respond is excluded, the corresponding figures were 272 and 14, respectively. Consecutive VEBs were seen in seven patients before DE was given and decreased significantly (p less than 0.05) 1.5-5.5 h after drug administration. There was no change in the PQ interval, the QRS interval showed a slight increase, whereas the QT interval was prolonged 0.5-4 h after administration of DE. A specific gas chromatographic method was used for DE assay in plasma and urine. Absorption was rapid in all patients. Urinary excretion during the first 48 h after drug intake varied between 35 and 75%. The lowest effective antiarrhythmic concentration estimated in six patients ranged from 1.4 to 7.0 mug/ml. beta-Phase half-life in five patients was between 10.3 and 22.1 h.


The hypothalamic syndrome in rats.

The hypothalamic syndrome in rats. The original conception of the hypothalamus controlling feeding by the activity of two specific and reciprocally inhibitory centers has now been largely abandoned. Detailed neural research using a wide variety of methods has demonstrated the complex morphological and functional organization of this part of the brain and has modified the earlier simplistic approach. However, examination of the feeding responses to a variety of stimuli that represent components of control of feeding indicates that much or even most feeding control is extrahypothalamic. As demonstrated by the obesity or aphagia resulting from hypothalamic damage or from reversible hypothalamic interference, the hypothalamus influences or modulates feeding control, possibly by an enabling action, but it does not itself substantially control food intake either in the short or the long term. In the cachaxia of cancer, which can tentatively be regarded as a negative obesity, and which is closely reproducible in a rat model, the decline of food intake can be attributed to failure of control components that are all extrahypothalamic, and the deterioration of control of feeding appears to be quite independent of the hypothalamus. The very detailed reconstruction of intrahypothalamic circuitry that has been developed in recent years has not yet had any real impact on the problem of where or how the active control of food intake is generated or the way in which the hypothalamus influences this control.


Gold thioglucose obesity syndrome.

Gold thioglucose obesity syndrome. Parenteral administration of gold thioglucose to mice produces an area or necrosis in the ventromedial portion of the hypothalamus. The lesion, like lesions produced by electrocautery of this area, causes hyperphagia and consequent obesity. The glucose moiety of gold thioglucose is essential for production of the lesion. Glucose analogues (2-deoxy-glucose, sodium thioglucose and phlorizin) prevent the gold thioglucose-induced lesion, and by themselves produce a transient hyperphagia. Insulin deficiency prevents the lesion. Either adrenalectomy or hypophysectomy counteracts the effect of insulin deficiency. Electron microscopic studies, in which general necrosis is avoided by administration of aspirin before gold thioglucose or by administration of subnecrotic doses of gold thioglucose, reveal that gold thioglucose primarily affects neural elements contiguous with capillaries in the ventromedial hypothalamus. The experimental observations indicate the presence of special glucoreceptor cells in the ventromedial hypothalamus that are involved in the regulation of food intake.


The Zucker-fatty rat: a review.

The Zucker-fatty rat: a review. The Zucker (fatty) rat is one of a group of animals that inherit obesity as an autosomal Mendelian recessive trait. These rats are obese, hyperphagic, and hyperinsulinemic, but blood glucose remains at normal levels. Although these rats eat more than normal rats, their response to the addition of adulterants to the food or after exposure to the cold is more like that of normal rats than rats with hypothalamic obesity. The hypertriglyceridemia which characterized these animals is due to the increased hepatic production of a very low density lipoproteins. Adipocytes are increased in size and in number with the subcutaneous fat depot showing the largest increase in the number of fat cells. Lipogenesis from glucose is brisk in the young animals but declines with age. Enzymatic patterns of glycolysis and gluconeogenesis appear to reflect the altered internal milieu rather than specific defects. Endocrine changes in the fatty rat include hyperinsulinemia, reduced levels of glucagon, hypothyroidis, and impaired reproductive function. A model is presented in which the features of the genetically obese (Zucker) fatty rat are compared with those of animals with hypothalamic obesity.


Gluconeogenesis: methodological approaches in vivo. The complexity of the process of gluconeogenesis makes it very difficult to study in vivo. Of the many approaches used to study this process, none have proven ideal. The arterial-hepatic venous catheterization technique can provide accurate determination of the net splanchnic uptake of the various gluconeogenic precursors but gives no information as to their fate once extracted. The infusion of a labeled precursor, such as [14C]alanine, will provide qualitative information about the fate of the labeled substrate but, without complicated kinetic analysis, will not provide quantitative information on the conversion of the precursor to glucose. Though each of these individual techniques has major limitations in the study of gluconeogenesis, various combinations of these methods can provide further insight into the regulation of this complicated process. By combining the arterial-hepatic venous catheterization technique with the infusion of a labeled precursor, the splanchnic extraction of the precursor can be measured as well as the amount of the extracted precursor that is converted to glucose. The concomitant infusion of two isotopes (a 14C-labeled gluconeogenic precursor to assess substrate conversion to glucose and 3H-labeled glucose to measure glucose turnover) can also provide an accurate determination of the amount of circulating precursor that is converted to glucose. The latter technique provides no information on the splanchnic extraction of the precursor but has the advantage of being less invasive and, therefore, more applicable to man. The last two techniques measure accurately the direct conversion of the circulating precursor to glucose and, therefore, provide reliable index of gluconeogenesis.


Stimulation of luteinizing hormone (LH) and follicle-stimulating hormone by (D-Leu6, des-Gly10-NH2)-LH-releasing hormone ethylamide after subcutaneous, intravaginal, and intrarectal administration to women. Women, most of whom had regular menstrual cycles, were administered D-Leu6,des-Gly10-NH2)-luteinizing hormone-releasing hormone (LH-RH) ethylamide (D-Leu6-LH-RH-EA)via different routes during the early or midfollicular phase of the cycle. Plasma LH, follicle-stimulating hormone (FSH), and estrogen levels were determined by radioimmunoassay before and after administration of D-Leu6-LH-RH-EA. plasma LH and FSH increased and reached peak levels 3 to 4 hours and 3 to 6 hours, respectively, after subcutaneous injection of 25 mug of the analog of LH-RH. Intravaginal or intrarectal application of 2 mg of D-Leu6-LH-RH-EA also increased plasma LH and FSH levels in most of the women, but the magnitude of the rise, the time of initiation of response, and the peak level varied among the women. The plasma estrogen level also rose after administration via either route.


Diftalone: a new non-steroidal anti-inflammatory agent: comparative study with phenylbutazone in the treatment of rheumatoid arthritis. A double-blind study was carried out to compare the effectiveness and tolerability of diftalone and phenylbutazone in thirty patients with classical or definite rheumatoid arthritis, randomly distributed between the two treatment groups. Both drugs were administered according to a progressively decreasing daily dosage schedule: 1,000 mg during the 1st week; 750 mg the 2nd week and 500 mg from the 3rd week on for diftalone; 400 mg, 300 mg, and 200 mg daily for the 1st, 2nd and from the 3rd week on respectively for phenylbutazone. The study lasted twelve weeks. The clinical controls and laboratory tests were performed weekly up to the 8th week, while the final evaluation was made at the end of the 3rd month. Twelve patients in the group receiving diftalone and fourteen in the phenylbutazone group completed the trial. Clinical improvement was observed in both groups. Effectiveness was somewhat more evident in the diftalone group. Tolerability was acceptable for both drugs, althoug the diftalone patients showed less frequent and intense side-effects than those treated with phyenylbutazone. No significant differences were found as regards the laboratory parameters, except a significant fall of the E.S.R. (less than 0-05) in the diftalone group. Diftalone seems to be an effective and safe anti-inflammatory agent in the treatment of rheumatoid arthritis.


An investigation into the effect on cigarette smoking of a new anti-smoking chewing gum. A preparation claimed to help patients to break the habit of smoking has recently been introduced. Material released in the mouth from a chewing gum causes an unpleasant taste when tobacco smoke is inhaled. This claim has been investigated in a double-blind trial on sixty subjects, thirty of whom took the active chewing gum and thirty the placebo chewing gum. The subjects each used one piece of chewing gum four times a day over a period of two weeks. This investigation clearly indicates that the active chewing gum is effective as an anti-smoking preparation, when used over a period of two weeks and the effect is still demonstrable one month later, although to a lesser extent.


Stimulated release of hypothalamic growth hormone-releasing activity by morphine and pentobarbital. Two minutes after the intravenous administration of 1 ml of sheep somatostatin antiserum in the rat, plasma growth hormone (GH) levels had reached a maximal 10-15-fold increase which remained approximately constant up to 90 min. The injection of sodium pentobarbital (2 mg/100 g body wt) led to a rapid and transient rise of plasma GH levels of similar magnitude (10-fold) reached 20 to 40 min after injection of the narcotic. The important finding is, however, that the combined administration of somatostatin antiserum and pentobarbital or morphine led to an almost exact additive effect on the plasma GH concentrations. These data, beside providing additional evidence for the existence of GH-releasing activity (GH-RH), indicate that morphine and pentobarbital stimulate GH secretion through increased release of GH-RH.


A comparison of pyrogenicity and related properties seen in a suspension of Corynebacterium parvum and a gramnegative organism. When the pyrogenic properties of C. parvum and a gramnegative organism, S. typhimurium, were compared it was found that the onset of the pyrogenic response to C. parvum was delayed relative to that of S. typhimurium and that secondary responses rarely occurred. In further experiments the interaction of C. parvum and S. typhimurium was studied. Although synergism was not demonstrated, the kinetics of the response to mixtures of the two vaccines was anomalous while pre-treatment of rabbits with C. parvum resulted in the elimination of the secondary pyrogenic response to S. typhimurium suspension. The implications of these results are discussed in terms of the known characteristics of endogenous pyrogen, lipopolysaccharide and peptidoglycan.


Factors affecting pyrogen testing in rabbits. A response of rabbits to toxins causing fever and originating from microorganisms may be related to different factors. In this respect the race and the age of the animals were examined as variables. Three rabbit strains of local origin were used: New Zealand White, White of Dendermonde and Holland race. As pyrogen preparations sterile surface water and preparations of E. coli strains were used. A bacterial count, as for ordinary drinking water examination, was carried out in order to evaluate the presence of bacteria to pyrogenicity. An attempt was made to remove the pyrogens by absorption on charcoal and by filtration through asbestos filters. A decrease of pyrogenicity was obtained but the fever toxins did not completely disappear with the simple procedure. The stability of the toxins found in surface waters was examined over a short range of preservation. Only a slow decrease in pyrogenic activity was seen. Pyrogens of E. coli were prepared in vitro, but used unpurified. A slight difference in the strains was observed, but all E. coli's were pyrogenic.


Effect of maternal vitamin A excess on S-100 in neonatal rat cerebellum: a preliminary study. Pregnant rats were administered a teratogenic dose of vitamin A excess on Days 17 and 18 of gestation and intubation controls received the vehicle alone. Pups were sacrificed at birth or on postnatal Days 5 or 10; the cerebellum was removed, frozen, and fixed in preparation for immunohistochemical localization of the brain specific protein, S-100. Vitamin A produced a transient delay in the appearance of S-100 in the external granular layer which was no longer evident on Days 5 and 10. These findings corroborate autoradiographic evidence that vitamin A temporarily interferes with neurogenesis. However, the early damage results in permanent behavioral deficits in adulthood despite apparent cytochemical repair.


The spontaneously diabetic Wistar rat. Metabolic and morphologic studies. A new, spontaneously diabetic syndrome has been recognized in nonobese outbred Wistar rats of both sexes. The age at detection of first glycosuria has varied from 48 to 120 days, with a mean of 67 days. Eighteen rats have been studied, 14 untreated and four during and after cessation of insulin treatment. The affected animals have demonstrated a spectrum of severity, with hyperglycemia (252-732 mg./dl.), hypoinsulinemia (0-1 ng./ml.), and hyperketonemia. The severely ketotic rats, with total blood ketone body levels between 6 and 13 mM, showed rapid loss in weight and dehydration over one to six days. The moderately ketotic (1-5 mM) declined gradually in weight over 15 days, with marked polyuria and glycosuria. The stable rats, with ketonemia less than 1 mM, sustained their weights, polyuria, and glycosuria for longer than 40 days. A relative or absolute increase in plasma immunoreactive glucagon and elevated levels of free fatty acids and branched-chain amino acids were observed in relation to the severity of the syndrome. Intraperitoneal arginine or tolbutamide elicited no insulin response, but the glucagon response to arginine was exaggerated. Pancreatic insulin content was normal or moderately decreased. Light-microscopic examination of pancreases of ketotic animals at the end stage of the disease showed islets to be very small and rare, consisting virtually of non-beta cells. In stable and earlier ketotic rats, the islets were small, with reduction in beta-cell number and a striking inflammatory cell infiltration. Surviving beta cells showed variable degranulation. This model of spontaneous diabetes in nonobese rats displays insulin deficiency, glucagon excess, and ketosis, with a dramatic inflammatory lesion during active beta-cell destruction.


Pancreatic islet-cell antibodies in diabetes mellitus correlated with the duration and type of diabetes, coexistent autoimmune disease, and HLA type. In a study of 972 patients with diabetes mellitus, humoral pancreatic islet-cell antibodies (I.C.Ab.) were detected in highest prevalence in insulin-treated diabetics with (38 per cent) and without (22 per cent) associated overt organ-specific autoimmune disease (A.I.D.) where consideration was not given to the duration of diabetes. They were also detected in 8 per cent of diabetics treated with oral hypoglycemic agents (O.H.A.), but not in diabetics requiring diet alone and in only 0.5 per cent of 434 control subjects. Six per cent of 522 patients with overt organ-specific A.I.D. but not diagnosed to be diabetic had I.C.Ab.s. I.C.Ab.s were present in the sera of 2 per cent of 157 first-degree relatives of I.C.Ab.-positive subjects. In insulin-treated diabetics and, to a lesser extent, in diabetics not requiring insulin, the prevalence of humoral I.C.Ab. was strongly dependent of the duration of the diabetes, being 60 per cent during the first year from diagnosis in the insulin-treated group and falling to 20 per cent at two to five years and to 5 per cent at 10-20 years. The prevalence of I.C.Ab. in insulin-treated diabetics showed no correlation with the patient's age at the time of testing when the duration of diabetes was taken into account. Diabetics who did not require insulin for treatment but who were I.C.Ab.-positive showed a significant tendency to subsequently require insulin and to have a higher prevalence of other autoantibodies than insulin-independent diabetics who were I.C.Ab.-negative. Persistence of I.C.Ab. for more than five years from diagnosis of diabetes was associated with coexistent overt organ-specific A.I.D. and with HLA-B8, A1, and A1 + B8.


Does insulin need a second messenger?

Does insulin need a second messenger? It is well established that specific binding sites for insulin are present on the plasma membranes of target tissues. In order to explain how insulin regulates a wide variety of biologic functions both on the surface of the cell as well as in its interior, it has been postulated that insulin generates a second messenger at the cell surface. To date, however, no second messenger for insulin has been identified that can carry out all of insulin's known actions. Recent studies have demonstrated that, in addition to the plasma membrane, other subcellular organelles, such as the nucleus, have specific binding sites for insulin. There is also evidence indicating that large serum proteins such as albumin, large protein hormones such as prolactin, and small protein hormones such as insulin can enter intact cells. It is hypothesized, therefore, that insulin has at least two mechanisms of action on target tissues. One mechanism entails the direct binding of insulin to the plasma membrane, which in turn leads to its well-known effects on membrane transport. The other mechanism requires the entry of insulin itself into the interior of the cell and its subsequent direct binding to subcellular organelles. This latter process then serves to mediate many of the known intracellular functions of insulin.


Long-term survival of adult rat islets of langerhans in artificial capillary culture units. Pancreatic islets from adult rats were cultured in artificial capillary cultury units that were perfused with nutrient medium. In this in-vitro culture system, islets remain functional for at least 97 days. As determined by insulin radioimmunoassay of the culture medium, islets continue to secrete insulin. Furthermore, islets responded to high glucose concentration in the perifusion medium by increased insulin release throughout the duration of the culture period. A normal appearance of individual epitheloid islet cells and the presence of aldehyde-fuchsin-positive granules were observed in islets fixed in medium. As indicated by the prolonged maintenance of tissue-specific function, the present simple technique for organ culture of intact islets should be useful for providing new information on the long-term effects in vitro of various factors on islet function and a means for the preservation of islets for transplantation.


Carbohydrate metabolism and pancreatic islet-cell function in thalassemia major. To investigate the development of diabetes mellitus in patients with thalassemia major, plasma glucose and immunoreactive insulin (IRI) levels following oral glucose and intravenous tolbutamide and glucose disappearance rates following intravenous insulin were measured in 10 patients before and during five years on a high transfusion program (HTP). Plasma immunoreactive glucagon (IRG) levels following oral glucose, intravenous insulin, and arginine were measured during the sixth year. Serial percutaneous liver biopsies were performed on seven patients. The oral glucose tolerance tests (OGAT) and mean peak IRI levels were normal in nine of 10 patients before HTP. After HTP was begun a progressive deterioration of OGTT occurred despite normal IRI levels. Following tolbutamide, the mean per cent fall in plasma glucose in the patients before HTP was significantly less than in controls (p less than 0.01) and similar to that of controls during five years of HTP in spite of higher than normal peak IRI levels. Of seven survivors after six years of HTP, three had normal OGTT and four had chemical diabetes; mean peak IRI levels were normal, but fasting IRG levels were significantly higher than in controls (p less than 0.05). In all seven patients, plasma IRG failed to increase following insulin-induced hypoglycemia and was significantly higher than in controls after arginine (p less than 0.01); after oral glucose, plasma IRG fell significantly below that of fasting only in the patients with chemical diabetes (p less than 0.03). Following intravenous insulin, the mean per cent fall in glucose before and during HTP was significantly less than in controls (p less than 0.01). Hemosiderosis and cirrhosis were present in all biopsied patients. Four patients died; two had chemical and two had nonketotic insulin-dependent diabetes. These data suggest that diabetes mellitus occurs frequently in patients with thalassemia on HTP and that insulin resistance and hyperglucagonemia, possibly due to cirrhosis, are important etiologic factors.