Biocompatibility tests of components of an implantable cardiac assist device. A permanently implantable in-series left ventricular assist device, the dynamic aortic patch (DAP), has been tested in chronic animal experiments. The DAP replaces a section of the intrathoracic aortic wall. Hemothorax and hematocele at the implantation site have been complications in recent experiments. Primary postoperative hemorrhage was ruled out, and the biocompatibility of all components was therefore examined. Dacron velour, Teflon felt, conductive polyurethane, segmented polyether polyurethane, and Teflon-coated polyester fiber sutures were implanted in the pleural cavities of dogs and tested in vitro by culturing canine saphenous vein explants on them. In vivo experiments demonstrated that all components elicited mild to moderate inflammatory reactions, but hematocele occurred only when the components were implanted in the aorta with direct blood contact and exposed to arterial blood pressures. In vitro, cells were cultured on all components with no signs of toxic reactions. These results indicated that the host tolerated all implant components without major inflammatory responses. However, histological data indicated that chronic slow bleeding into or through the Dacron velour in contact with the arterial blood serum could account for hemothorax or hematocele formation. Therefore, a configuration of the assist device using materials impermeable to blood may obviate these difficulties.


Isolation and characterization of the membrane envelope enclosing the bacteroids in soybean root nodules. The membrane envelope enclosing the bacteroids in soybean root nodules is shown by ultrastructural and biochemical studies to be derived from, and to retain the characteristics of, the host cell plasma membrane. During the early stages of the infection process, which occurs through an invagination, Rhizobium becomes surrounded by the host cell wall and plasma membrane, forming the infection thread. The cell wall of the infection thread is degraded by cellulolytic enzyme(s), leaving behind the enclosed plasma membrane, the membrane envelope. Cellulase activity in young nodules increases two- to threefold as compared to uninfected roots, and this activity is localized in the cell wall matrix of the infection threads. Membrane envelopes were isolated by first preparing bacteroids enclosed in the envelopes on a discontinuous sucrose gradient followed by passage through a hypodermic needle, which released the bacteroids from the membranes. This membrane then sedimented at the interface of 34--45% sucrose (mean density of 1.14 g/cm3). Membranes were characterized by phosphotungstic acid (PTA)-chromic acid staining. ATPase activity, and localization, sensitivity to nonionic detergent Nonidet P-40 (NP-40) and sodium dodecyl sulfate (SDS) gel electrophoresis. These analyses revealed a close similarity between plasma membrane and the membrane envelope. Incorporation of radioactive amino acids into the membrane envelope proteins was sensitive to cycloheximide, suggesting that the biosynthesis of these proteins is primarily under host-cell control. No immunoreactive material to leghemoglobin antibodies was found inside or associated with the isolated bacteroids enclosed in the membrane envelope, and its location is confined to the host cell cytoplasmic matrix.


Compositional analysis of growing axons from rat sympathetic neurons. We describe culture systems for neurons of an adrenergic autonomic ganglion which: (a) permit cultivation of neurons without supporting cells, (b) permit separate harvest of somal and axonal material, and (c) permit direct access to the neuronal surface. The antimetabolites used to suppress supporting cell growth did not have demonstrable effects on neuronal polypeptide synthesis. Rapid neurite outgrowth, which characterized these cultures, was prevented by colchicine or cycloheximide and resumed promptly after their withdrawal. Axons separated from cell bodies showed no incorporation of label from leucine or fucose, but did exhibit incorporation of glucosamine. The major polypeptides present in this neuron, as demonstrated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis, are described. No major differences in polypeptide content were observed when soma and axons were compared. Likewise, there were no differences detected in polypeptides synthesized by neurons in suspension or neurons actively extending processes. Analysis of the polypeptides within the neurites after labeling with amino acids indicated transport at a number of different rates; certain of these polypeptides corresponded in size and transport characteristics to polypeptides observed in the rabbit optic nerve after labeling of retinal ganglion cells. Tubulin and actin have been definitively identified in this cell type (18); we found proteins similar in size and proportionate amounts to be among the rapidly transported soluble polypeptides. The prominent polypeptides observed after several methods of surface labeling are described.


Use of type-specific antimyosins to demonstrate the transformation of individual fibers in chronically stimulated rabbit fast muscles. Continuous stimulation of a rabbit fast muscle at 10 Hz changes its physiological and biochemical parameters to those of a slow muscle. These transformations include the replacement of myosin of one type by myosin of another type. Two hypotheses could explain the cellular basis of these changes. First, if fibers were permanently programmed to be fast or slow, but not both, a change from one muscle type to another would involve atrophy of one fiber type accompanied by de novo appearance of the other type. Alternatively, preexisting muscle fibers could be changing from the expression of one set of genes to the expression of another. Fluorescein-labeled antibodies against fast (AF) and slow (AS) muscle myosins of rabbits have been prepared by procedures originally applied to chicken muscle. In the unstimulated fast peroneus longus muscle, most fibers stained only with AF; a small percentage stained only with AS; and no fibers stained with both antibodies. In stimulated muscles, most fibers stained with both AF and AS; with increasing time of stimulation, there was a progressive decrease in staining intensity with AF and a progressive increase in staining intensity with AS within the same fibers. These results are consistent with a theory that individual preexisting muscle fibers can actually switch from the synthesis of fast myosin to the synthesis of slow myosin.


Studies on the nature of protease-induced growth stimulation in normal and transformed BHK cells. In the presence of growth-limiting serum concentrations trypsin displays mitogenic activity on actively-growing but not quiescent BHK cells. These results suggest that BHK cells arrested in G1 (G0) are not sensitive to protease-induced growth stimulation. Previous work strongly suggested that the trypsin active-site is not directly involved in its mitogenic activity on BHK cells. Additional studies on denatured trypsin fragments further indicate that the molecular conformation and size of native trypsin may not be absolutely required for mitogenic activity. Cellular multiplication induced by the addition of fresh serum to quiescent BHK cultures is not inhibited by high concentrations of soybean trypsin inhibitor. Similar to our previous findings with trypsin, it has been further observed that plasmin is not sufficient to initiate the growth of BHK cells in soft agar. Trypsin also fails to enhance the growth of a thermosensitive polyoma-transformed BHK line in soft agar at the restrictive temperature. Finally, the growth of transformed BHK cells in soft agar does not display a requirement for plasminogen and is not inhibited by soybean trypsin inhibitor. These studies argue against the involvement of plasmin or other exogenous trypsin-like enzymes in the growth and transformation of BHK cells.


Ultrastructural basis for the assay of topical acne treatments. Transmission and scanning electron microscopy of untreated comedones. After two decades of using systemic antibiotics as a main treatment for acne, emphasis is again being placed on topical agents. Thus, it is highly desirable to have a procedure whereby the activity of the various compounds can be evaluated by direct visualization. Scanning electron microscopy combined with transmission electron microscopy provide the tools for such in assay. This study describes the ultrastructure of untreated comedones and provides the baseline or control data necessary for testing topical treatments. Comedones obtained by punch biopsies or comedo extractors were processed for electron microscopy and studied with an ETEC Autoscan Scanning Electron Microscope and with Philips EM 300 and EM 301 Transmission Electron Microscopes. Microorganisms, keratinized cells, sebum, and hairs interact with each other to form the comedonal mass. Corynebacterium acnes and Pityrosporum ovale proliferate abundantly in close association with sebum and penetrate the keratinized cells. Fine structural details of bacteria and yeasts as well as features of host-microbial relationship have been elucidated.


Dermabrasion by diamond fraises revolving at 85,000 revolutions per minute. Use of a high-speed, air-driven turbo-grinder equipped with diamond fraises that revolve at 85,000 revolutions per minute has improved results in dermabrasions on the face by, it is estimated, 50%. Dermabrasions have been done with this turbo-grinder for scarring from acne, actinic damage, syringomata, lesions of adenoma sebaceum and some other facial conditions. The rapid evaporation, nonimflammability, nonnarcotic, and excellent freezing properties of Freon 12 eliminate the necessity for auxiliary cooling fans. It is thought too that the incidence of secondary infections and keloids have been decreased by use of an ointment of erythromycin (1%) topically during the healing period.


A survey of the visual and developmental-perceptual abilities of the Down's syndrome child. This pilot project was designed to survey the visual and developmental-perceptual abilities and capabilities of the Down's syndrome child. Considering the frequency of Down's syndrome in the general population, there has been a paucity of work done in this area. Each child was given a complete visual evaluation using standard pediatric techniques and careful objective observations of performance. Subjective techniques were used whenever possible, however, subjective testing could not be used on many of the subjects due to problems concerning their overall comprehension of the task and their ability to converse, as many infants were included in this study. All subjects were also given a developmental-visual examination which is composed of tests with readily available normative data, so that subjects could be compared to each other and to normal children.


[Ocular symptoms in pituitary adenomas (author's transl)]. The study of 85 patients suffering from pituitary adenoma, amongst which 56 with a visual failure, allowed to show: 1) The importance of the functional loss a revealing sign of the disease (50% cases). 2) The importance of the ophthalmological examination as a diagnostic argument. The latter, however, needs an extremely methodical examination of the visual fields. The combinated use of the Goldman Perimeter, and the Amsler schemes gave us satisfactory results: 87.5% of patent bitemporal hemianopsiae. It has been useful, in several cases to enhance these methodes by using coloured indexes. All of the patients underwent surgery, with complementary radiotherapy. The functional recovery quality has been shown, in the majority of the cases, to be proportionnal to the operation precocity in relation to the first functional symptoms.


[Prolapse of the iris through spontaneous perforation of the cornea and pathological costal fracture after cortisone therapy for rheumatoid arthritis (author's transl)]. A case of spontaneous corneal ulcer with iris prolapse is reported in a patient suffering from rheumatoid arthritis. The concomitant presence of a pathological costal fracture, recognised to be steroid nature, suggests a correlation between the corneal manifestations and the corticosteroid therapy to be established.


[Bilateral hereditary corneal dermoid (author's transl)]. The opportunity to follow the descendants of a family carrying the defect of a bilateral corneal lesion already described in the literature has led the authors to recommend surgical treatment and the lesions have been studied by light and electron microscopy. The two diagnoses are discussed: scleocornea and dermoid. They retain the latter diagnosis and give their reasons.


[Actual treatment of rhabdomyosarcoma of the orbit in children (author's transl)]. Among malignant tumor of the orbit in children, rhabdomyosarcoma remains the most frequent; it should be suspected on any unilateral exophthalmia. Twenty cases have been treated in the "Institut Gustave Roussy" (1960--1975). It can be shown with polychimiotherapy and a better coordination in timing of the other therapies, the survival rate improved clearly. In addition, since 1972 more localized forms have been treated. In the future, the survival rate is supposed to improve yet, if the diagnosis is made earlier. In such conditions a recovery with few inconveniences may be expected. Exenteration is an important mutilation and do not supply a better protection. A secondary inucleation due to complications of radio-therapy is not an exception. However a prothesis will be possible later.


[Macular dystrophy in menkes disease. Histological ocular study (author's transl)]. Pathological ocular study of a boy presenting Menkes disease and died at age 5 months showed rarefaction of outer nuclear layer and degenerative visual cells' outer segments in macular area.


[Orbital sarcoidosis (author's transl)]. In case of an isolated orbital tumour, which evolves rapidly in a patient, whom general condition is good and who shows no other signs, the possibility of a sarcoidosis must be considered. The diagnosis will be confirmed by a biopsy and often by a tomography of the pulmonary hila, a radiography of the hands and feet as well as by the efficacity of the corticosteroid treatment.


"Tie-in" trapezium prosthesis: Long-term results. Seventeen of 20 thumbs treated with a silicone-Dacron trapezium replacement arthroplasty for disabling rheumatoid and osteoarthritis at the base of the thumb were reviewed. Patients were uniformly satisfied with the results. Relief of pain was achieved in all but three patients. The latter required excision for tissue reaction to the implant. (Histological findings are reported in these cases.) Motion and stability were satisfactory but not normal. Postoperative subluxation occurred in six cases. Wear and bone erosion were present in two cases after operation.


Impairment of host vs graft reaction in pregnant mice. I. Suppression of cytotoxic T cell generation in lymph nodes draining the uterus. The fetus resulting from an allogenic (interstrain) mating represents a type of graft that is not rejected by the mother. Nevertheless, the maternal immune system seems to recognize and to react to the presence of the fetus in a number of ways. One such manifestation is significant enlargement of the lymph nodes that drain the uterus (DLN) of pregnant rodents. We have tested the DLN lymphocytes of mice for reactivity to paternal H-2 alloantigens after interstrain mating. The DLN lymphoid cells obtained from pregnant mice killed fewer newborn F1 recipients in a graft-vs-host mortality assay, and generated less cytotoxic T cell activity against paternal H-2 antigens both in vivo and in vitro. In vitro mixing experiments demonstrated the presence of a cell-associated suppressor activity in the DLN of pregnant mice. This suppressor proved resistant to treatment with mitomycin C, and appeared in the DLN early in pregnancy.


In vitro growth of cytotoxic human lymphocytes. I. Growth of cells sensitized in vitro to alloantigens. Human lymphocytes sensitized to allogeneic determinants in vitro were continuously cultured on medium prepared from phytohemagglutinin-(PHA-P) stimulated human mixed lymphocyte cultures. With such human-conditioned medium (HCM), human peripheral lymphoid cells could be grown in culture for over 90 days with a doulbing time of about 54 hr. Cytotoxic lymphocytes could be grown in culture for this time with no loss of cytotoxicity or change in cytotoxic specificity.


Effects of penicillamine and hydroxyquinoline on absorption of orally ingested 65zinc in the rat. The influence of high doses of 2 chelating agents, D-penicillamine and iodochlorohydroxyquin, on the metabolism of orally ingested 65 Zn was studied on rats by means of whole body counting assay technique. Both drugs were found to increase zinc absorption in a dose-dependent way to values significantly above those of the untreated control group. At identical doses D-penicillamine had a significantly stronger enhancing effect than the 8-hydroxyquinoline derivate, except for the highest dosage which was 100 mg daily. The turnover of the absorbed 65Zn fraction was followed during the time of observation which was 11 days. The elimination rate was practically identical in all experimental groups, except for the 100 mg D-penicillamine group in which the biological half-life of 65Zn was significantly decreased as compared to the control group.


The phage adsorption test: a simple method for differentiation between staphylococci and micrococci. A simple phage adsorption test is recommended for differentiation between staphylococci and micrococci. All 132 strains of Staphylococcus tested inactivated phage U16 significantly, whereas of 123 strains of Micrococcus tested, only three strains of Micrococcus candicans were able to adsorb this staphylococcal phage. There are special problems in the taxonomy of M. candicans.


Nonproducer malignant tumor cells with rescuable sarcoma virus genome isolated from a recurrent Moloney sarcoma. Cells from a secondary tumor developing at the site of a regressed Moloney sarcoma virus-induced tumor could be passaged in adult STU mice by intramuscular and intraperitoneal inoculation. The tumors induced by these cells, as well as by a cell line derived from it, grew progressively and led to death of the animals between 3 and 7 wk after tumor transplantation. No evidence for production of virus from these cells was obtained or for the presence of viral antigens (p30, gp69/71). From both cell variants, sarcoma virus genome could be rescued by infection with helper virus, resulting in the establishment of a cell line producing focus- and XC plaque-forming virus. The rescued producer cells very frequently also produced tumors which finally grew progressively. The nonproducer cells were not immunogenic, as was demonstrated in cross transplantation tests and in studies for cell-mediated cytotoxicity (CMC) and complement-dependent antibody-mediated cytotoxicity (AMC). The producer cells, however, were demonstrated to possess a strong immunogenicity. The nonproducer cells, though nonimmunogenic, revealed a weak immunosensitivity when used for challenge in the transplantation protection assay or as target cell for the demonstration of AMC and CMC, if the immune response was induced by cells producing the sarcoma-helper virus complex, but not by cells producing only helper virus. The nonproducer cells, as well as their rescued producer derivative, showed a stronger reactivity with cytotoxic antibodies than with cytotoxic cells, whereas the helper virus-producing cell line was comparably suitable as target cell for AMC and CMC. The recurrence of a regressed Moloney sarcoma is assumed to be the result of the occurrence of transformed nonproducer cells escaping immune destruction, and not as a consequence of a depleted immune resistance in the host.


Characteristics of Ca2+- and Mg2+-induced tension development in chemically skinned smooth muscle fibers. Chemically skinned fibers from guinea pig taenia caecum were prepared by saponin treatment to study the smooth muscle contractile system in a state as close to the living state as posible. The skinned fibers showed tension development with an increase of Ca2+ in the solution, the threshold tension occurring as 5 X 10(-7) M Ca2+. The maximal tension induced with 10(-4) M Ca2+ was as large and rapid as the potassium-induced contracture in the intact fibers. The slope of the pCa tension curve was less steep than that of skeletal muscle fibers and shifted in the direction of lower pCa with an increase of MgATP. The presence of greater than 1 mM Mg2+ was required for Ca2+-induced contraction in the skinned fibers as well as for the activation of ATPase and superprecipitation in smooth muscle myosin B. Mg2+ above 2 mM caused a slow tension development by itself in the absence of Ca2+. Such a Mg2+-induced tension showed a linear relation to concentrations up to 8 mM in the presence of MgATP. Increase of MgATP concentration revealed a monophasic response without inhibition of Ca2+-induced tension development, unlike the biphasic response in striated muscle. When MgATP was removed from the relaxing solution, the tension developed slowly and slightly, even though the Mg2+ concentrations was fixed at 2 mM. These results suggest a substantial difference in the mode of actin-myosin interaction between smooth and skeletal muscle.


Hereditary optic atrophy with probable association with a specific HLA haplotype. A family with hereditary optic atrophy in 4 members of 3 generations is described. The clinical findings differ from those previously observed in hereditary optic atrophy and in Leber's disease, indicating that the hereditary optic atrophy described in this report represents a new disease entity, which is inherited in an autosomal, dominant way with incomplete penetrance. Analyses of the cerebrospinal fluid and serum did not reveal signs of immunoglobulin synthesis within the central nervous system, i.e. findings encountered in a considerable proportion of patients with optic neuritis. An association was found between the family members affected by the disease and the major histocompatibility system haplotype A2 B8. A linkage thus occurred between the disease and the HLA region on human chromosome No. 6.


Simultaneous determination of niacin, niacinamide, pyridoxine, thiamine, and riboflavin in multivitamin blends by ion-pair high-pressure liquid chromatography. A high-pressure liquid chromatographic procedure for the simultaneous determination of niacin, niacinamide, pyridoxine, thiamine, and riboflavin was developed and applied to the analysis of multivitamin blends for these water-soluble vitamins. Reversed-phase ion-pair chromatography, using sodium hexanesulfonate as the counterion, was employed. Analysis time is shortened considerably and precision is improved by the application of this analytical technique as compared to the current official methods of analysis. Involved sample pretreatment is not required.


Endotoxin fever in the new-born guinea-pig and the modulating effects of indomethacin and p-chlorophenylalanine. 1. At 30 degrees C ambient temperature E. coli endotoxin injected into the cerebral ventricles evokes a febrile response in 0-3 day-old guinea-pigs. If the dose is sufficiently high, the fever is biphasic: two rising phases separated by a transient fall.2. At 20 degrees C ambient temperature the change in body temperature after the endotoxin is still biphasic, but the transient fall is more pronounced and, finally, hypothermia develops. The relatively large surface area of the new-born cannot explain, by itself, the hypothermia.3. The phasic changes in body temperature following endotoxin administration are unlikely to be mediated by a single central factor, and a sequence of several factors could be postulated.4. Indomethacin prevents the first-phase febrile rise in body temperature, and also the consequent fall, but not the second-phase rise.5. p-Chlorophenylalanine pre-treatment prevents the transient fall only, it slightly increases the first-phase rise and does not influence the second-phase rise.6. Prostaglandins and/or other derivatives of endogenous arachidonic acid in the brain might be responsible for the first rising phase of the endotoxin fever, and might also initiate a central serotonergic mechanism which, in turn, could lead to the transient falling phase between the two rising phases of fever. The mechanism of the second-phase febrile rise in body temperature awaits some other explanation.


Advantages of cryopreserved lymphocytes for sequential evaluation of human immune competence. II. Mixed lymphocyte cultures and mononuclear cell subpopulations. Peripheral blood lymphocytes were obtained from normal volunteer donors over a 2-week period. One-way or two-way mixed lymphocyte reactions, percentages of T-cells and B-cells, and percentages of monocytes were quantitated on both fresh and cryopreserved lymphocytes. These comparisons revealed that the measurement of mixed leukocyte reactions, fractions of E-rosettes and EA rosettes, and the fraction of esterase-staining cells were not altered by cryopreservation and storage at -196 degrees C up to 1 year. Cryopreserved reference lymphocytes provided a stable standard for use as stimulating cells in mixed lymphocyte cultures and could be irradiated or treated with mitomycin C prior to being frozen, without alteration of their function as stimulator cells. Cryopreserved cells thus appeared to be utilizable in the sequential measurement of the immune competence of humans.


Multiple myeloma: an immunologic profile. II. Bone marrow studies. Bone marrow mononuclear cell populations were studied in 35 patients without myeloma, 39 patients with multiple myeloma, and 15 patients with benign monoclonal gammopathy. Bone marrow mononuclear cell receptors, responses to mitogens or allogeneic stimuli, and suppressive effects on in vitro peripheral blood lymphocyte (PBL) function were studied. In bone marrow cell populations from patients with untreated multiple myeloma, the percent of complement receptor-bearing cells and the pokeweed mitogen- and concanavalin A-stimulated responses were significantly greater than were those in bone marrow cell populations from patients without myeloma. Sheep red blood cell receptor-bearing cells were significantly greater in marrow populations from treated multiple myeloma patients compared to those from untreated multiple myeloma patients. Sheep red blood cell receptor-bearing cells from the bone marrow of multiple myeloma patients suppressed responses of the multiple myeloma patients' PBL's to autologous mitomycin C-treated bone marrow plasma cells and to allogeneic stimuli in one-way mixed leukocyte culture. Complement receptor-bearing cells suppressed the response to pokeweed mitogen. The presence of lymphocytes in the marrow compartment that are capable of suppressing the response of myeloma patients' PBL's to plasma cell antigens may be significant in the pathogenesis of multiple myeloma.


Down's syndrome and thyroid dysfunction. A child with Down's syndrome (DS) and associated hyperthyroidism is described and studies pertinent to DS and thyroid function are discussed. A majority of data supports a hypothesis of immunological abnormalities in DS, with resultant thyroid dysfunction. It is puzzling that overt disease is not more common in DS, since the incidence of thyroid autoantibodies is high and autoimmunity and Hashimoto's thyroiditis are often associated with clinical thyroid disease. A physician who cares for children should consider thyroid disease in any child with DS who manifests a single compatible symptom. Palpation of the thyroid gland is essential during each physical examination.


Studies of the cardiac sarcoplasmic reticulum in myocardial infarction. 1. Influence of ischemia on the biochemical properties of the sarcoplasmic reticulum (SR) was studied in the experimental myocardial infarction in the dog. 2. Ca2+ -uptake rate of SR decreased at around 90 minutes after coronary occlusion. This reduction was roughly in parallel with the reduction in the Ca+ -Mg2+ -stimulated ATPase activity. However, Ca2+ -binding rate of SR was kept within the range of that of the non-infarcted tissue through the time course of myocardial infarction. 3. Ca2+ -Mg2+ -stimulated ATPase activity decreased at around 3 hours after coronary occlusion to about 50% of that of the non-infarcted portion. 4. In SDS gel electrophoresis, the protein band with the largest molecular weight among three major components decreased at 3 hours after coronary occlusion, which is suggestive of ATPase. At 48 hours after coronary occlusion, the protein with the smallest molecular weight in the major proteins also decreased. 5. Ca2+ -uptake rate, Ca2+ -Mg2+ -stimulated ATPase activity and the substructural changes return to the normal level and pattern at around 28 days after coronary occlusion.


Interaction between noradrenergic and serotonergic mechanisms on the central regulation of blood pressure in the rat: a study using experimental central hypertension produced by chemical lesions of the locus coeruleus. The interaction between noradrenergic and serotonergic mechanisms on the central regulation of blood pressure in the rat was studied, using central experimental hypertension produced by chemical lesions of the locus coeruleus (LC) with 6-hydroxydopamine (6-OHDA). The hypertension (LC-hypertension) was blocked by pretreatment with 6-OHDA intraventricularlly administered and also with desipramine. An increase of serotonin (5-HT) turnover in the cortex and especially in the spinal cord was revealed in the hypertension. Intraperitoneally administered para-chlorophenylalanine elevated blood pressure and reduced 5-HT and 5-hydroxyindole acetic acid content in the cortex and especially in the spinal cord, indicating a decrease 5-HT turnover. Intraventricularlly administered 5, 6-dihydroxytryptamine (5, 6-DHT) resulted in only bradycardia but not an elevation in blood pressure and did not block the development of LC hypertension, and lowered 5-HT content in the cortex and especially in the spinal cord. Intraspinally administrated 5, 6-DHT evoked mild but significant elevation in blood pressure but not in heart rate, and lowered 5-HT content only in the spinal cord. Furthermore, the disappearance of the LC was histologically observed in the rats with LC hypertension. The results of these experiments demonstrate that (1) LC hypertension is probably due to a specific lesion of the LC with 6-OHDA, (2) LC hypertension is accompanied by an increase of 5-HT turnover in the cortex and especially in the spinal cord, which may participate in a depressor function in LC hypertension, (3) ablation of bulbospinal serotonergic neurons produce mild elevation in blood pressure. (4) the balance between the activity of central noradrenerfic and serotonergic neurons may play a role in maintaining normotension, and their unbalance may induce hypertension.


The eradication of Encephalitozoon cuniculi from a specific pathogen-free rabbit colony. Encephalitozoan cuniculi was discovered in a large specific pathogen-free rabbit colony during routine quality assurance testing. By using a modified India-ink immunoreaction test we were able to test the entire colony for antibodies to Encephalitozoon cuniculi. The prevalence of the disease was approximately 5%. All seropositive animals were culled, and another test of the entire colony, carried out 4 weeks later, revealed one seropositive rabbit which was also culled. Two subsequent screenings of the whole colony have shown no further seropositive animals.


Immunological methods employed in an attempt to induce erythema nodosum leprosum (ENL) in mice. In the mouse foot pad model five different parameters were employed to simulate the condition of ENL as observed in the human. The experimental groups with five to six months leprosy infection were injected intravenously with various anti-mycobacterial antibodies, M. leprae sonicate and 'B' cells obtained from syngenic donors. The control group of animals, infected similarly, were treated either with M. leprae sonicate or gamma globulins precipitated from normal human serum. All recipients were killed at one day interval excepting those with 'B' cell transfer, which were sacrificed at seven days. The sites of predilection for immune complex deposition viz. foot pad and kidney as well as the spleen were examined by light and fluorescence microscopy. These findings are described and discussed.


[Characteristics of nucleoside and pyrimidine base metabolism in Corynebacterium glutamicum]. Corynebacterium glutamicum ATCC 13032 can utilize uridine as a sole source of carbon for its growth. Uridine degradation seems to be catalysed by hydrolase since it does not depend on the presence of phosphorus when the enzyme activity is assayed in vitro. Other nucleosides (adenosine, inosine, guanosine, cytidine, thymidine, deoxyuridine, deoxyadenosine, deoxyguanosine) cannot serve as a source of carbon for the growth of the bacterium. The label of thymidine-3H (methyl) is not incorporated into the acid-insoluble fraction of nucleic acids whereas the label of thymine-14C is incorporated. The cells are not susceptible to the presence of fluorodeoxyuridine in the medium but are inhibited by low concentrations (0.01 mcg/ml) of fluorouracil, the inhibition being eliminated with equimolar concentrations of uracil. Therefore, Corynebacterium glutamicum cannot assimilate thymidine or its analogue, fluorodeoxyuridine, in contrast to thymine which can be used as a sole source of nitrogen in the medium.


Differential transcription of nonrepeated DNAs in Neurospora crassa mutants. Transcriptional properties of an "albino" mutant, a "slime" mutant and a wild type strain of Neurospora crassa have been compared by quantitative estimates of DNA:RNA hybridization. The RNAs isolated from these strains were incubated with 32P-DNA of the wild type strain. The DNA:RNA hybrids were isolated by chromatography on hydroxyapatite. 29--34% of the wild type 32P-DNA hybridized with RNA from all of the strains used. The DNA sequences which gave a primary reaction with wild type RNA (tr-DNA) were recovered and again reacted with different RNAs. The tr-DNA sequences reacted to the extent of 92% with the wild type RNA, but only 78% with slime mutant RNA and 85% with the RNA isolated from the albino mutant. The use of tr-DNA sequences thus isolated, proved to have increased the specificity of the hybridization reactions, these were useful indicators for identification of transcriptional differences among wild type and mutant strains of N. crassa.


The human plasma fibrinolytic system: regulation and control. The factors involved in the regulation and control of the human plasma fibrinolytic system at the cellular level are unknown at this time. The physiological regulation of plasmin formation in plasma depends primarily on the nature of the circulating zymogen, plasminogen, the physiological activators formed both in the blood and in the vascular endothelium, and the specific plasmin inhibitors found both in plasma and in certain of the cellular elements of the blood. The biosynthesis of the zymogen must be under genetic control, and the activators are probably released, after thrombus and clot formation, from components involved in the surface-mediated initiation of the coagulation system, and from the vascular endothelium. Activation of plasminogen can occur both in the fluid phase surrounding the thrombus and probably at thrombus surfaces, involving both the fibrin clot and the platelet membrane. The plasmin inhibitors act to control the system in order to prevent proteolytic degradation of important physiologic trace proteins of the coagulation, complement and kallikrein-kinin systems by the enzyme.


Spontaneous deaths of fetuses with chromosomal abnormalities diagnosed prenatally. In a survey of pregnancies, in which a chromosomal abnormality was diagnosed by amniocentesis but in which the mother did not undergo an abortion, the spontaneous fetal death rate after amniocentesis for 21 fetuses with Down's syndrome was 24 per cent (or 21 per cent, considering 19 singletons only). This rate is significantly greater than the rate of 2.7 per cent among 73 fetuses with less seriously abnormal genotypes reported in this survey or of 3.3 per cent and 3.5 per cent in the Canadian and United States studies of amniocentesis. After midtrimester there is about a sixfold higher risk of spontaneous fetal death for a fetus with Down's syndrome than for one with a normal genotype. At least some (and probably a major fraction) of the discrepancy observed between maternal-age-specific rates of Down's syndrome in live births and those found after amniocentesis is due to spontaneous fetal loss. A similar inference may be drawn for trisomy 18.


An immunofluorescence study of the passive arthus reaction in rat sciatic nerve. The Arthus reaction, passively induced in rat sciatic nerve by local injection of antibody and intravenous injection of antigen, was studied by immunofluorescence, using fluorescein and tetramethylrhodamine isothiocyanate. The reaction in nerve is similar to that occurring in skin. Antigen-antibody complexes formed at the site of the reaction activate complement and attract large numbers of polymorphonuclear leucocytes which then ingest the immune complexes. The significance of the Arthus reaction in relation to diseases of the peripheral nervous system is mentioned.


[Number of malarial mosquitoes and their feeding hosts under urban and rural conditions]. In connection with building of hydrotechnical constructions and creation of water bodies the abundance of anopheles as in urban so in rural conditions was high that amounted to 120 to 690 females per a site of day's rest. By means of precipitation reaction in the capillaries with an extraction of stomachs' contents it has been established that in rural conditions hosts of anopheles are horned cattle while in urban ones--man and poultry.


[Anomalohimalaya lotozkyi sp. n., a new species of ixodid ticks from the Peter the First Range (Ixodoidea, Ixodidae)]. Anomalohimalaya lotozkyi sp. n., the second species of this genus, was found in Tadzhikistan, Peter I ridge, at the height of 2700 m above the sea level in rocky locality with xerophilous vegetation on Alticola argentatus. In June, 1977 there were found larvae and nymphs of the species. After feeding larvae and then nymphs females and males were obtained. All phases of the life cycle differ distinctly from those of the type species An. lama Hoog., Kais. et Mitch., 1970, from Himalayas. The diagnosis of the genus Anomalohimalaya is specified. This genus combines the characters of Haemaphysalis Koch and Rhipicephalus Koch and is of great significance for more precise definition of phylogeny of the subfamily Amblyomminae Banks.


[Experimental study of the role of Scarabaeus sacer beetles in the eliminaton of Thysaniezia giardi (Cestoda: Avitellinidae) eggs]. Experimental studies have shown that when feeding beetles of S. sacer swallow eggs of Th. giardi together with faeces of horned and small cattle. After getting into the insect's intestine the eggs of Thysaniezia are eliminated in most beetles (from 91.1 to 100%) within one hour. Only in some individuals a very insignificant portion of the parasite's eggs remains viable within two hours. In beetles' faeces excreted by them in two hours and in subsequent periods after the feeding eggs of Thysaniezia were not found. The greatest eliminational ability was displayed by the beetles maintained without feeding for 1--2 days. The eliminational ability of beetles kept over 3 days (4 to 5) without food noticeably decreased.


[Fleas from small forest mammals of the Udmurt ASSR]. 578 fleas of 16 species were found on 3518 animals collected in coniferous-broadleaved forests in the vicinity of the town of Izhevsk. Am. penicilliger, Lept. silvatica and H. talpae are most abundant. The main host of the fleas is the common redbacked vole. In addition to 15 species of the ectoparasites characteristic of rodents and insectivorous animals of this region Cer. garei (a bird flea) was found on the common redbacked vole.


[Pleistophora ladogensis sp. n., a microsporidian (Protozoa, Microsporidia) from the musculature of the burbot, Lota lota, and of the smelt, Osmerus eperlanus eperlanus ladogensis]. The parasite develops inside the fibres of the cross-striated musculature. On the histologic sections of the affected tissue most young stages of P. ladogensis are represented by 2--6 nucleic cells 5--6 mu in size. Mature pansporoblasts have a thick and solid membrane. Their average size is 43 (18--60) mu. The number of spores in pansporoblasts is over 16. Live spores are egg-shaped and are 5.4 (5.0--5.8).2.9 (2.7--3.3) mu in size. The length of the polar filament is 100 to 180 mu.


Resolution of the membrane moiety of the H+-ATPase complex into two kinds of subunits. The H+-translocating ATPase complex from the thermophilic bacterium PS3 (TF0-F1) is composed of a water-soluble part with ATP-hydrolyzing activity (TF1) and a membrane moiety with H+-conducting activity (TF0). TF0 was obtained by treating TF0-F1 with urea and removing contaminations on a carboxymethyl-cellulose column. This TF0 contained only two kinds of subunits, band 6 protein (13,500 daltons) and band 8 protein (5400 daltons), and it was active in H+ conduction and TF1 binding when reconstituted into proteoliposomes (TF0 vesicles). The binding of TF1 to TF0 present in vesicles restored energy-transducing activities, such as ATP-32Pi exchange, dicyclohexylcarbodiimide-sensitive ATPase, and ATP-dependent enhancement of 8-anilinonaphthalene-1-sulfonate fluorescence. Treatments such as protease digestion and chemical modification with acetic anhydride, succinic anhydride, or diazobenzenesulfonic acid destroyed the TF1-binding activity, which was caused by band 6 protein. Band 8 protein was a proteolipid that reacted specifically with dicylcohexyl-carbodiimide and seemed to play a central role in H+ conduction through the membrane.


Interaction between actomyosin and 8-substituted ATP analogs. Various 8-substituted ATP analogs were synthesized, and their reactions with myosin and actomyosin were studied. The nucleoside triphosphates (NTPs) with an amino group at the 6 position and hydrogen at the 8 position, and formycin 5'-triphosphate (FTP) were hydrolyzed by myosin very slowly in the presence of Mg2+ and rapidly in the presence of EDTA and K+. In contrast, NTPs with substitution of the 8 position, other than FTP, were readily hydrolyzed by myosin in the presence of Mg2+ but were hardly hydolyzed in the presence of EDTA and K+. The Michaelis constant (Km) for hydrolysis of 8-substituted NTP by heavy meromyosin was much larger than the dissociation constant (Kfl) for binding of heavy meromyosin with NTP estimated from the change in tryptophan fluorescence. All the NTPs with no substitution at the 8 position, and FTP, caused an initial Pi burst, actin activation of myosin NTPase, superprecipitation of actomyosin, and myofibrillar contraction. On the other hand, all the 8-substituted NTPs in three possible conformations did not cause these phenomena, regardless of the conformation. These results were discussed in relation to the hindrance of rotation about the glycosidic bond accompanying an 8 substitution.


Effects of para-chlorophenylalanine and 5-hydroxytryptophan on mouse killing behavior in killer rats. The effects of para-chlorophenylalanine (PCPA) on mouse killing behavior were examined in natural killer rats. Forty-eight hr after injection, this serotonin synthesis inhibitor, at relatively low doses of 75 and 150 mg/kg, facilitated mouse killing, as indicated by a decrease in latency to attack the mouse. This effect was revealed in a test of satiation, in which five successive mice were presented to the rat, and also in a novel cage situation. Other than the shorter latencies to attack and kill mice, the killing response was similar in topography to the natural kill. The increase in killing after PCPA injection was associated with a reliable reduction in brain serotonin and in 5-hydroxyindoleacetic acid, and the time courses of the behavioral and biochemical changes were generally similar. In contrast to PCPA, injection of the serotonin precursor 5-hydroxytryptophan (5-HTP, 100 mg/kg) reliably lengthened attack and kill latencies in killer rats. In rats pretreated with PCPA, 5-HTP not only reversed this drug's facilitation of killing, but completely blocked killing in 67% of the rats tested. These results strengthen the hypothesis that brain serotonergic neurons are involved in the inhibition of mouse killing.


Characterization of inhibitor(s) of lymphocyte activation in serum from rats with adjuvant arthritis. Serum from adjuvant arthritic rats inhibits the concanavalin A- (Con A) and lipopolysaccharide-induced stimulation of lymph node cells, leaving the basal and phytohemagglutinin-stimulated 3H-thymidine incorporation unaffected. Con A-stimulated 3H-thymidine uptake is also inhibited in rat spleen and peripheral blood lymphocytes and in dog peripheral blood lymphocytes. The intensity of the inhibitory activity in serum is positively correlated with the intensity of the secondary lesions of adjuvant arthritis. Inhibitory activity was not found in serum from rats bearing nystatin-induced inflammation. Serum fractionation studies indicated that the inhibitory activity cannot be attributed to low molecular weight alpha2-glycoproteins or to gamma-globulins and alpha2-macroglobulins, but it is present in a fraction migrating with beta-globulins. The inhibitory activity in arthritic rat serum is reduced by treatment with non-steroidal anti-inflammatory drugs, but is unaltered by D-penicillamine. It is suggested that this inhibitory activity is part of the systemic response to an immunologically mediated inflammation.


[Histological, electromicroscopical and histochemical studies on the central nervous system of rats after chronic treatment with imipramine (author's transl)]. Rat brains have been studied after treatment with oral doses of 50 mg imipramine/day for 3 and 6 months. 20 brains have been studied histologically, 3 brains electronmicroscopically, 6 brains histochemically as well as 34 controll brains. On the light microscopic level no pathologic changes of intravital origin have been revealed. The hyperchromatic changes of neurons were of the same character and degree and showed the same topic distribution in the experimental and in the control group. They should be regarded as postmortem artifacts. The pyramidal cells of hippocampus field h3, the Purkinje cells and the Golgi epithelial cells have been examined by electron microscopy. Besides a possible slight induction of lysosomes no alterations could be found. The histochemical studies (succinate dehydrogenase, ATPase, AMPase, acid phosphatase, PAS, methylgreenpyronin) revealed no differences between the experimental and the control group.


Prenatal ultrasonographic diagnosis of fetal gastroschisis. Two cases of fetal gastroschisis were diagnosed in utero by ultrasound. In one patient, a cluster of sonolucent "cystic" structures was seen in the amniotic fluid which corresponded to fluid-filled bowel loops. The second infant was the firstborn of dichorionic twins and the gastroschisis abnormality appeared as two sonolucencies in relationship to the trunk. An extensive review of the literature is presented.


[Resection-arthrodesis of the knee for bone tumours (author's transl)]. The authors describe the technique, devised by Juvara and modified by Merle d'Aubigné of resection-arthrodesis of the knee with fixation by medullary nailing and using massive bone grafts. They report 38 cases operated on for various conditions (20 giant-cell tumours, 7 chondro-sarcoma, 6 juxta-cortical sarcoma, 3 other types of sarcoma, one desmoid fibroma, one chondroma). The best results were obtained in cases in which surgical excision had been sufficiently wide and when very thick nails were used. The use of homografts increased the rate of infection. Resections of the lower and of the femur resulted in more complications than tibial resections. In femoral lesions, the part of the tibia that is transplanted should not be turned upside-down but should be mobilised upwards to provide better bone contact. The indications are discussed and compared with those of total replacement. The authors believe that, despite the number of complications, this technique is more practicable than total knee replacement.


[Clear-celled chondrosarcoma or chondroblastic sarcoma. A new type of chondrosarcoma (author's transl)]. The authors report five cases treated by wide resection. Pathological studies were made of the whole of the lesion. Three tumours were situated at the upper end of the femur, one at the upper end of the humerus and one at the level of the tibial plateau. The specific features were localisation in an epiphysis and histological areas of clear cells whose cytoplasm contained glycogen. From the anatomical and radiological points of view, these tumours were like chondroblastomata but their progress was more like that of chondrosarcomata. The author concludes that they should be distinguished from both types of tumour.


Clinical investigations of neocarzinostatin in Japan. Neocarzinostatin (NCS) is an antibiotic from streptomyces carzinostaticus which inhibits DNA synthesis. Clinical trials in Japan began in 1971. NCS is active against S-180, Ehrlich tumor, L1210, Yoshida sarcoma, and a range of ascitic hepatomas. In rabbit NCS is distributed at high concentrations in the kidney, skin, stomach, pancreas, lung, and muscles. The high distribution in the pancreas and the stomach suggested possible effectiveness in human tumors at these sites. In clinical studies NCS has been shown to be active against acute leukemia. As a single agent 9 out of 51 obtained a CR with 9 more achieving a PR. Anorexia, nausea, and vomiting were the most frequent side effects. NCS has been tried in combination with Ara-C, daunorubicin and prednisolone and CR was ssen in 11 out of 14. In stomach cancer responses of some kind were observed in 12 out of 141 cases, while in the case of pancreatic tumors there were 10 out of 68.


Comparative mutagenicity and toxic effects of 2,5,2',5'-tetrachlorobiphenyl and its metabolites in bacterial and mammalian test systems. The possible toxicological properties of 2,5,2',5'-tetrachlorobiphenyl (TCB), 4-hydroxy-2,5,2',5'-tetrachlorobiphenyl (TCB-OH), and 2,5,2',5'-tetrachlorobiphenyl-3,4-oxide (TCB-oxide) were evaluated in three bioassay systems. Incubation of rat hepatocyte plasma membranes with 50 microgram/ml of TCB-OH decreased the activity of Mg2+-ATPase by approximately 90%, whereas approximately half of the enzyme activity remained after the TCB or TCB-oxide treatment. All three compounds were found to be inactive in the Salmonella/microsome mutagenicity assay. Only TCB-OH possessed potent cytotoxic activity against all four S. tymphimurium strains tested. It affects the viability of TA 1537 by as much as 40% even at concentrations as low as 1 microgram/ml. These data suggest the potential toxicological significance of metabolic activation by the hepatic microsomal mixed-function oxidases.


F1-hybrid anti-parental-strain reactivity. I. Impaired proliferation of CBA lymphocytes in the spleens of irradiated C3H X CBA mice pretreated with CBA spleen cells. Previous investigations have indicated that C3H X CBA hybrid lymphocytes can neutralize lymphocytes which are reactive against the C3H-determined Mls antigen when injected into the H-2-compatible strain CBA. In this study we have inoculated C3H X CBA hybrids with CBA spleen cells and 2 weeks later lethally irradiated the mice and injected them with CBA lymphocytes to study their proliferation in the host spleen. Proliferation of lymphocytes from CBA mice, but not from C57Bl mice, was significantly impaired in such hosts, indicating that the hybrids had become immunized against the parental cells. In contrast, such an effect was not detected by treating CBA X C57Bl mice with CBA spleen cells. Humoral antibodies in the C3H X CBA hybrids capable of blocking the response of CBA lymphocytes against C3H X CBA cells could not be detected, indicating that inhibition of lymphocyte proliferation is cell mediated rather than by humoral factors.


F1-hybrid anti-parental-strain reactivity. II. Proliferation of C3H X CBA hybrid lymphocytes in the spleens of irradiated CBA mice. Experiments were conducted to explore whether C3H X CBA lymphocytes can react by proliferation when they meet parental CBA spleen cells. MLC tests could not detect such a reaction, but infusion of C3H X CBA lymph node cells into irradiated CBA hosts resulted in rapid cell proliferation in the host's spleen. Such a cell proliferation was also observed after infusion of T-cell-enriched lymph node cell preparations or thymic cells from C3H X CBA donors. Upon transfer to new irradiated mice these proliferating cells continued to proliferate in CBA mice but not in C3H X CBA mice. Further evidence that the injected T-cells were the proliferating emerged from experiments where AKR X CBA lymphocytes were found to proliferate in spleens of irradiated CBA mice and that most of these cells posessed the theta antigen determined by AKR. Proliferation of F1-hybrid lymphocytes in the spleens of its irradiated parental strains was found not to be a general phenomenon and is probably restricted to some Mls-antigen-compatible strain combinations. The possibility that the C3H X CBA hybrid lymphocytes are stimulated to proliferations by CBA lymphocytes reactive against the C3H-determined Mls antigen is discussed.


Polymorphonuclear leucocyte function during pregnancy. Polymorphonuclear leucocyte (PMN) function was studied in pregnant women and related to the serum concentration of pregnancy zone protein (PZP) and to mixed lymphocyte culture (MLC) reactivity. Neutrophil chemotaxis was depressed in pregnant women. Pregnancy serum inhibited the MLC-reaction. The serum levels of PZP were inversely related to chemotactic responsiveness (P less than 0.05) and depression of MLC reactivity (P less than 0.01). The capacity to reduce nitroblue tetrazolium was depressed in PMNs from pregnant women, and pregnancy serum inhibited phagocytosis of Escherichia coli by control PMNs. Neutrophils from pregnant women showed increased chemiluminescence during phagocytosis of zymosan. The results may be explained by depression of ingestion by pregnancy serum and increased oxidative metabolism in PMNs from pregnant women.


What is interstitial fluid? Biochemical and physiological analysis of fluid obtained from tissue cages. Multiperforated polypropylene balls implanted in soft-tissue and renal parenchyma deliver fluid which represents interstitial fluid, which was characterized biochemically and physiologically. In a time-concentration study, the serum antibiotic level was compared to levels in interstitial fluid of the kidney (RIF), interstitial fluid of soft-tissues of the abdominal wall (STIF) and urine, measured simultaneously after intravenous injection of six different antibiotics. A direct time-concentration relationship between the concentration of antibiotics in the serum and in the renal interstitial fluid was found. This study helps delineate the basic pharmacokinetics in renal interstitial tissue and suggests guidelines for optimal therapy in renal parenchymal infections.


The relationship between physical disability, social perception and psychological stress. It was hypothesized that the prevailing attitudes towards the disabled would affect their self-concept and thus the impact of the disability as a stress stimulus. This hypothesis was examined in two separate studies. The first study evaluated the impact of disability context on the evaluations of disabled individuals by the non-disabled. It was found that disabled veterans were evaluated significantly higher than individuals disabled in traffic, work accidents, or as a result of polio. In the second study, the self-concept of disabled veterans was compared to the self-concept of work accident disabled. The disabled veterans had significantly higher self-concept than those of the people disabled in work accidents. The results would appear to demonstrate the relationship between the variables presented in the hypothesis.


Relation between the low back pain syndrome and x-ray findings. 2. Transitional vertebra (mainly sacralization). The relation between LBP and transitional vertebra was investigated in 312 subjects; 148 healthy individuals served as controls. An overall incidence of 0.65% lumbarization and 20.8% sacralization was found; of the latter, one third were partial and two-thirds complete. The incidence was markedly lower in women. No direct relation between sacralization, partial or complete, and LBP was found; this was based on interrelation of the LBP and control groups, with or without sacralization, to age, ethnic community, occupation, number of changes of occupation or place of employment, duration of LBP, and physical occupational requirements (sitting, standing, bending, sudden maximal effort). There is some evidence that LBP, when associated with sacralization, may be more severe.


[Treatment of acne and seborrhea using antiandrogens]. 57 women suffering from seborrhea and acne were treated with low doses of estrogens and antiandrogens. A cyclic regimen comprising 21 tablets of 0.05 mg ethinylestradiol and 2 mg cyproterone acetate was used. Results were very favourable: within 6 months, 51 patients were healed or significantly improved. There were only minor side effects, and none of the women conceived. It is concluded that this combination not only provides very effective therapy but might also be the contraceptive of choice for women with skin problems.


Interposition mesocaval shunt for bleeding esophageal varices: clinical, hemodynamic, and metabolic changes. Twenty-three patients bleeding from varices had mesocaval H-grafts using Dacron prosthetic material. There were 21 men and two women with ages ranging from 37 to 60 years. There were 11 elective and 12 emergency procedures. Five patients (17%) were classified as Child's class A, seven (34%) as class B, and 11 (49%) as class C. An operative mortality of 17% was noted in the entire group, with one late death due to hepatic failure. All deaths occurred in emergency cases belonging to class C. Technical difficulties were encountered more often in the hands of less experienced surgeons. A high rate of shunt patency and minimal problems with postoperative encephalopathy were noted despite abnormal results of ammonia tolerance tests postoperatively. Lack of hepatopedal flow was noted postoperatively compared with preoperative angiograms. There were no serious adverse effects from diversion of blood from the liver in our surviving patients. The interposition mesocaval shunt is a technically safe and easy procedure in the hands of experienced surgeons. We continue to recommend the mesocaval shunt until other methods prove better than this operation.


Epidemiological features of skin diphtheria infection in Rangoon, Burma. A survey for skin diphtheria was carried out on 493 patients with skin lesions who were attending Veneral Diseases & Dermatology Clinic in Rangoon in 1971. 63.8% of the patients had C. diphtheriae in their skin lesions. All ages and both sexes were equally affected. Skin lesions yielding C. diphtheriae on culture were indistinguishable from those associated with other bacteria including ecthyma, pyoderma and secondarily infected with parasitic infections. Out of 108 strains tested 18.5% were found to be toxigenic, mainly intermedius. No toxigenic gravis strain was encountered. No relation existed between the type of lesion and the type or toxigenicity of the infecting strain. 75% of these toxigenic strains were isolated from children under 5 years of age. The phage types of toxigenic strains isolated were: II-IV, XII, Ph r, 8, 19-20, III, IVII and IV.


Activities of hepatic enzymes in spontaneous diabetes rats produced by selective breeding of normal Wistar rats. Acitivites of the hepatic enzymes were determined in spontaneous diabetes rats. The activities of the enzymes were compared with those in normal rats and in streptozotocin diabetic rats. In the spontaneous diabetes rats, glycogen phosphorylase and glycogen synthase were 14.6 +/- 0.6 and 1.73 +/- 0.15 U respectively. The activities of both the enzymes were significantly increased. In the spontaneous diabetes rats glucokinase was 3.82 +/- 0.5 U showing a significant increase. On the contrary, the activity of the enzyme was decreased in the streptozotocin diabetic rats. Glucose-6-phosphatase was increased both in the spontaneous diabetes rats and in the streptozotocin diabetic rats. Fructose-1,6-diphosphatase was increased in the spontaneous diabetes rats. Glucose-6-phosphate dehydrogenase was increased in the spontaneous diabetes rats and decreased in the streptozotocin diabetic rats. In the spontaneous diabetes rats phosphofructokinase showed a reduction of the activity and glucose-6-phosphate dehydrogenase was elevated. These findings are consistent with the results of activities of the hepatic enzymes in adult-onset diabetic patients. These patterns of the hepatic enzymes in the spontaneous diabetes rats were different from those in the streptozotocin diabetic rats. From these patterns of activities of the hepatic enzymes, the spontaneous diabetes rats produced by repetition of selective breeding according to Goto et al. (1975,1976) are an excellent model of human adult-onset diabetes.


Serological matching for D locus antigens improves kidney allograft survival in rhesus monkeys. A prospective study performed in unrelated, unimmunized rhesus monkeys revealed that serological matching for D-associated Ia1 antigens improves kidney allograft prognosis, provided that the recipients are nonreactive in mixed lymphocyte cultures (MLC) against their donors. MLC-negative host-donor combinations showed a more than 3-fold increase in the mean survival time as compared with MLC-positive combinations. Apparently, compatibility for the most important MLC-stimulating determinants is required for a significant improvement in graft survival. On the other hand, matching for A and B loci antigens did not play a decisive role. Since the relatively simple matching on the basis of identity for Ia1 (or DR) antigens yields a rather high proportion of MLC-nonreactive combinations, this method is likely to become an essential addition to the conventional matching procedures, also in clinical organ transplantation.


Correlation of tissue typing, mixed lymphocyte culture, and related donor renal allograft survival. Results of mixed lymphocyte culture reactions and tissue typing were correlated with the clinical courses of recipients of living related donor renal allografts. Forty-nine patients tested by the mixed lymphocyte culture technique were divided into two response groups: stimulation index greater than 5 and stimulation index less than 5. Seventy patients were tested by standard tissue typing methods and were categorized by the number of misstimulation in mixed lymphocyte culture than with mismatched antigens, suggesting that lymphocyte-defined histocompatibility is more important than serologically defined histocompatibility in selecting the best possible allograft donor.


Inhibition of cell-mediated microcytotoxicity and stimulation of mixed lymphocyte reactivity by mouse pregnancy serum. Blocking activity of mouse pregnancy serum was investigated using a well defined one-way mixed lymphocyte culture and a cell-mediated microcytotoxicity test. Allogeneic (C57BL (H-2b) X CBA/Ca (H-2k)) and syngeneic (C57BL (H-2b) X C57BL (H-2b)) pregnancy and postpartum sera were found to stimulate rather than inhibit the mixed lymphocyte response. In contrast, the same sera were able to abrogate target cell killing by hyperimmune lymphocytes in a cell-mediated microcytotoxicity test. Although inhibitory activity was present in both allogeneic and syngeneic 18-day pregnancy sera, indicating a non-specific effect, only allogeneic postpartum sera had significant blocking activity when compared with either the equivalent syngeneic group or the normal serum control. The specificity implied by this gained support from preliminary third-party experiments. Blocking activity was found in primiparous as well as multiparous sera. These results provide further evidence for the existence of maternal serum-blocking factors which may operate to protect the conceptus against maternal cell-mediated immunity during pregnancy, and indicate that they may act at the efferent rather than the afferent phase of the immune response.


In vitro immunosuppressive effects of cytotoxic agents conjugated to antihuman lymphocyte globulin. Antihuman lymphocyte globulin (ALG) was either coupled to the lymphocytoxic drug chlorambucil or covalently bound to the cytotoxic alkalating agent melphalan via a polyglutamic acid carrier. Both types of complexes strongly inhibited the proliferative response in human mixed lymphocyte cultures and the ability of mixed lymphocyte culture-activated T effector cells to lyse 51Cr-labelled lymphoblast target cells, and were more potent than ALG or drug alone. These experiments indicate that it is possible to bind cytotoxic agents to ALG without destroying either the properties of the drug or the ability of the antibody to bind to lymphoid cells.


Modulation of mixed lymphocyte culture reactivity following alloimmunization between H-1 (Ag-B)-histocompatible rat strains. H-1 (Ag-B)-histocompatible rats were cross-immunized to study the effects of presensitization on the mixed lymphocyte culture (MLC) reactivity in normally nonresponsive MLC interactions. Lewis (LEW) rats, H-1 (Ag-B), were given s.c. injections of Fischer (F), H-1 (Ag-B), lymhocytes at weekly intervals for 3 weeks. Immunized LEW (Lew-iF) and F lymphocytes showed increased proliferative responses in MLC after the third immunization. The augmented proliferative responses in the MLC between these H-1 (Ag-B)-histocompatible rat strains following immunization may reflect minor histocompatibility antigenic differences. On the other hand, decreased proliferative responses in MLC between Lew-iF and Ag-B (H-1)-disparate rat strains were demonstrated. Alloimmunization across minor histocompatibility differences led to different types of regulatory effects on the MLC interactions. With low MLC responses, e.g., minor histocompatibility differences, the regulation tends to be positive; whereas for strong MLC responses, e.g., major H-1 (Ag-B) differences, the regulation tends to be negative or suppression.


Immune response-associated antigens on mouse leukemia cells. II. Anti-Ia sera inhibit the MLR reaction between normal GR spleen cells and syngeneic spleen cells of GRSL tumor-bearing mice. Mitomycin C-treated ascites cells of Ia antigen-positive GRSL14 tumor cells and spleen cells from GRSL14 tumor-bearing mice stimulated lymphocyte proliferative responses in normal syngeneic GR spleen cells. Furthermore, mitomycin C-treated T cells purified from spleen cells of tumor-bearing mice also stimulated normal GR spleen cells. Anti-Ia sera inhibited the stimulating ability of tumor cells and of spleen cells of tumor-bearing mice. These data suggest a role for I region gene products in immune surveillance for syngeneic tumors.


Immune reactivity of cells from long-term rat renal allograft survivors. Lewis rats receiving an LBN kidney allograft demonstrate no signs of rejection if they are pretreated with donor spleen cells and antiserum reactive with the donor alloantigen. We examined the cellular reactivity of long-term kidney allograft survivors. Normal proliferative and cytolytic responses were obtained with spleen cells from long-term survivors, in marked contrast to the diminished responses of cells from neonatally tolerant rats or the heightened cytolytic response of cells from rats that had rejected a renal allograft. Serum from long-term renal allograft survivors as well as serum obtained from rats at the time of transplantation did not suppress proliferative or cytolytic responses of normal cells. The results of this study suggest that long-term renal allograft survivors possess the precursors of those cells which are responsible for proliferative and cytolytic responses in mixed leukocyte cultures, but that they have not been sensitized to their renal allograft.


[Effect of gene dose in human cells].

[Effect of gene dose in human cells]. Data from literature are reviewed on the gene dosage effect in human cells. Until recently the dosage effects have been distinctly shown for the genes of superoxide dismutase-1, acid phosphatase of erythrocytes, adenine phosphoribosyl transferase, glutathione reductase and nucleoside phosphorylase, located on chromosomes 21, 2, 16, 8, 14, respectively. The data are discussed in the light of problems arising from the study of the regulation of gene expression in man.


Computer analysis of micturition parameters and accuracy of uroflowmeter. Several micturition parameters in healthy volunteers have been studied with digital computer analysis. The accuracy of the uroflowmeter has also been tested. Total voiding time, average flow rate, maximum flow rate, volume and time at maximum flow rate are all the function of volume voided. The first half of urinary volume is rapidly evacuated in the first one third of the total voiding time, and the rest in the remaining two thirds of the voiding period. When the constant stream hits at the center of the disk, the response of the flowmeter is fairly good. Once the urinary stream is accelerated and/or hits at the edge of the disk, the response is not reliable.


Subcellular B cell calcium and insulin secretion in vitro. Comparative ultracytochemical studies after glucose stimulation and cyproheptadine inhibition. Using the ultracytochemical pyroantimonate technique different patterns of calcium containing precipitates were found in the B cells of the isolated perfused rat pancreas under conditions of stimulated and inhibited insulin secretion. The calcium specificity of the ultracytochemical method was assessed by perfusion with a EGTA containing calcium-free medium, which markedly reduced the extent of precipitation. Perfusion with 20 mM D-glucose over a period of 30 min resulted in calcium distribution patterns which could be related to the biphasic insulin release. The calcium patterns differed significantly in their quality and quantitative morphometry from those after 5 mM D-glucose or cyproheptadine (CPH) perfusion (20 mM D-glucose plus 0.1 mM CPH). After 3--5 min of 20 mM glucose perfusion there was an increased calcium precipitation along the inner side of the B cell membranes. After 20--30 min an additional increase in precipitation was found in the cytoplasmic matrix and in the secretory granules. B cells in a CPH-inhibited state of secretion and also after perfusion with 5 mM glucose lacked these findings. The data suggest that an increase in the membrane associated calcium may induce the first phase of insulin secretion by triggering the exocytosis of peripheral granules, while the cytoplasmic calcium may be involved in long term regulation of insulin release.


An effective morphometric method for electron microscopic studies on papillary muscles. Morphometry was performed on the left ventricular posterior papillary muscles of seven Wistar rats. The volume densities of myocardial cells, interstitial space, myocardial nuclei, sarcoplasm, mitochondria, myofibrils, ground substance and T tubules, and the surface densities of myocardial cells, mitochondrial membranes and T tubules, were calculated. Though only 1 ultrathin section per animal was evaluated the low standard errors of the means indicate that the method described here will be adequate in most experimental studies. Due to the anisotropy of the surfaces within myocardial cells, the papillary muscles were cut at an angle of 32.4 degrees to their longitudinal axis. This angle is derived from an equation published by Whitehouse (1974). The procedure to correct the loss of cristal membrane images from oblique sectioning is discussed.


An ultrastructural study of the placenta in materno-fetal rhesus incompatibility. An electronoptical study has been made of eleven placentae from cases of materno-fetal rhesus incompatibility. The characteristic findings are focal, but sometimes quite extensive, syncytial necrosis, retention of normal pinocytotic and secretory activity in the non-necrotic syncytiotrophoblast, cytotrophoblastic hyperplasia, thickening of the trophoblastic basement membrane, immature-type endothelial cells in the fetal villous vessels and thickening or lamination of the capillary basement membranes. The pathogenesis of many of these chantes is not clear but there is no evidence that they are immunologically mediated. It is suggested that the syncytial necrosis may be due to narrowing of the intervillous space as a result of increased villous size, that the cytotrophoblastic hyperplasia is a response to the syncytial damage and is responsible for the changes in the trophoblastic basement membrane and that the fetal capillary changes are indicative of endothelial cell damage due, possibly, to fetal anaemia. Despite the damage suffered by the placenta in materno-fetal rhesus incompatibility there is little evidence of impaired functional efficiency.


Studies on subcutaneous fat necrosis of the newborn. Biopsy specimens from the skin and subcutaneous fat tissue of four cases with neonatal subcutaneous fat necrosis were made and investigated by light and electron microscopy at 2, 4, and 6 weeks, and 5 months (Case 2) from the onset of the disease. Three stages of ultrastructural change of fat cells were observed. The evolution of crystal formation in the fat cells was seen and phagocytosis of crystals and fat droplets by macrophages and foreign-body giant cells was also noted. In the light microscope accumulation of calcium concretions in the spaces between and inside the fat cells was found. In the electron microscope we detected foci of highly electron-dense granules, which were similar in distribution and structure to calcium salts stained with the von Kossa method. Changes in small and medium size blood vessels were observed.


Chordoid sarcoma of the soft tissue of the nape of the neck: a case with a 20 year follow-up. A huge tumor mass arising from the soft tissue of the nape of the neck, which histologically resembled chordoma is reported. This case had a history of over 30 years after onset at around 37 years of age. With frequent recurrence and re-excision after en bloc radical resection, performed at 47 years of age, this female patient expired at the age of 66, 20 years after the first radical operation. At operation, the mass was found to be firmly attached to the dorsal soft tissue. There was no connection with any bony structure, such as the cervical vertebrae or skull. Chordoma-like histological features were unchanged throughout the next 20 years. This unusual malignant neoplasm appears to correspond to the "chordoid tumor" or "chordoid sarcoma" of soft tissue, a term coined by F.W. Stewart in 1948. Alternatively the name "chondroid chordoma" used by Heffelfinger, Dahlin and others may be employed.


Heat injuries to the respiratory system. A steam-tube of the main boiler exploded on a ship lying in the harbour of Hamburg. The steam temperature was 283 degrees C. Cutaneous and severe inhalational scalding occured in the 27 fatalities, the men dying after different intervals. This paper deals with the pathological findings in the respiratory passages and the lung, describing the topographical extent of direct thermal injury and the temporal course of tissue reactions. In cases of instantaneous death coagulation necrosis of the tracheal and bronchial wall was found to extend to alveolar ducts in central parts of the lung. The lung parenchyma showed marked congestion, alveolar edema and desquamation of alveolar epithelial cells. Death occured due to acute pulmonary dysfunction and shock. Lethal complications following the period of primary shock consisted of fulminant confluent bronchopneumonia, the hyaline membrane syndrome or the onset of desquamative interstitial pneumonia. These changes rendered it difficult to evaluate the effects of the heavy cutaneous scalding on the pathological course of inhalational injuries in those surviving for longer periods.


Double outlet right ventricle in a human embryo. A human embryo of 14 mm crown-rump length is described to exemplify the anomaly of double outlet right ventricle with subaortic defect. The configuration of endocardial swellings in the outflow portion of the heart are compatible with the general architecture of full-grown examples of the anomaly. This configuration differs considerably from normal embryonic anatomy, but is still supported by the architecture of the myoepicardial mantle. It is concluded that endocardial swellings play a secondary role in the determination of fullgrown cardiac anatomy.


[Pinguecula and pterygium: histologic and electron microscopic study (author's transl)]. Two pingueculae combined with pterygia were studied by light and electron microscopy. Hyaline degeneration of the collagen, dark staining granular, von Kossa negative concretions and elastotic material were observed in both conditions together with marked changes in the fibroblasts, endothelial cells, pericytes and the basement membrane of conjunctival capillaries and small veins. The elastotic material is similar to that observed in solar elastosis, where the collagen fibers are less severely damaged. Chronic sun exposure of the pericorneal conjunctiva may damage endothelial cells primarily and disturb vascular exchanges. This would result in accelerated degeneration and regeneration of endothelial cells, in thickening of the basement membrane and, secondarily, disturbed metabolism of fibroblasts with alterations of the collagen and elastic fibers.


[Investigations on the ultrastructure of lympho-epithelial thymomas with special reference to "emperipolesis" (author's transl)]. The ultrastructure of eleven thymomas with lymphocytic predominance, one "epitheloid" cell thymoma and two normal human thymuses is described with special reference to "Emperipolesis". All patients have had myasthenia gravis. The normal human thymus consists of three parts: outer cortex, inner cortex, and medulla. The outer cortex contains mainly lymphoblasts and Metcalf's macrophages within the so-called "Clark-packet's". The inner cortex consists mainly lymphocytes and interdigitating reticulum cells, and the medulla of epithelial cells, lymphocytes and Hassall's corpuscles. In all cases of lympho-epithelial thymoma and in normal human thymuses there are enormous interdigitations between epithelial (tumor) cells, lymphocytes and macrophages. The "epitheloid" cell thymomas also show findings which suggest an epithelial cell interaction. We have not found intact lymphocytes inside the cytoplasm of normal and/or tumor epithelial cells, macrophages or interdigitating reticulum cells. The intracellular existence of intact lymphocytes has been termed "Emperipolesis" by Humble, Jayne, and Pulvertaft, meaning "internal wandering". These investigations indicate that "Emperipolesis" is not an adequate term for cellular interaction in normal human thymuses and thymomas. A false impression of intraepithelial location of thymic lymphocytes is created by two-dimensional sections of complex thymic structure. These ultrastructural studies revealed damage to lymphocytes only in macrophages with lymphocytolysis within these cells and accumulation of numerous heterophagic vacuoles containing fragments of lymphocytic debris within them.


Primary fibroxanthosarcoma of the thoracic aorta. A case is presented of a 58-year-old female with a primary malignant tumor of the aorta. The lesion was diagnosed histologically as a fibrox-anthosarcoma and represents the first primary neoplasm of the aorta so classified. The gross and microscopic features of the lesion are described and the morphologic characteristics of previously reported aortic sarcomas are briefly reviewed  and examined in light of the current histologic criteria for soft tissue tumors.


The effect of the epidermal G2 chalone on the mitotic duration in nude mouse epidermis and in a transplanted squamous cell carcinoma. Balb/c/nu nude mice transplanted with a moderately differentiated squamous cell carcinoma were injected intraperitoneally with different doses of aqueous skin extracts containing the epidermal G2 chalone. The mitotic counts and the mitotic rates were determined in histological sections using a stathmokinetic method with vinblastine sulphate. The mitotic duration was calculated from the mitotic rates and counts. Skin extracts containing epidermal G2 chalone increased the mitotic duration in the epidermis, and a similar trend was seen in the tumour. The higher the dose of chalone, the longer the mitotic duration tended to be. A straight line of best fit used to indicate the dose/response relationship was steeper for the epidermis than for the tumour. The study thus shows that the epidermal G2 chalone not only prevents epidermal cells from entering mitosis, it also prolongs the mitotic duration. Further, the results do not contradict the theory that tumour cells may be less sensitive to chalone than normal cells.


Cumulative injury.

Cumulative injury. A few states, notably California, are experiencing large increases in the number and cost of disability settlements under workers' compensation. Claims of cumulative injury for coronary heart disease, hypertension, stroke, cancer and neuropsychiatric problems have all been interpreted as compensable under workers' compensation, even when these conditions are clearly related to the aging process. Legal precedents for such claims are building rapidly throughout the country. The resultant costs may lead to the demise of the workers' compensation system. The situation in California is discussed in detail including the legal aspects, cumulative injury claims by type of disease and age of claimants, legal costs to the individual and the employer, and the economic outlook for the workers' compensation insurance system.


[The importance of the endo-endothelial fibrin and mucopolysaccharide film for the vessel wall permeability]. After a short description of the morphology and the present ideas of the function of the endo-endothelial fibrin and mucopolysaccharide film, respectively, of the vascular wall hitherto undertaken experimental investigations of this structure within the pathogenesis of the arteriosclerosis are summarized. The results of these investigations are contradictory. The difficulties of a morphological demonstration of changes of the endo-endothelial film going along with disturbances of the permeability as an early change of the arteriosclerosis are in the method of demonstration, in which case it is not sure, whether with the preparative and technical means used at present processes taking place on molecular level in the substances of the endo-endothelial film may really be comprehended.


[Fibrin deposits and fibrinolysis in pre and early stages of arteriosclerosis]. Already in the early stages of atherosclerosis still before the appearance of coarsely visible changes subendothelially fibrin-like material is found in the places of predilection known, which reveals electron-microscopically the typical periodicity. Lesions of the endothelium with increase of permeability, release of coagulation factors and mitogenic substances which stimulate the proliferation of smooth muscle cells always precede. There are several mechanisms for the penetration of fibrinogen or fibrin into the vascular wall, the proportion of which changes with the progressing of the atherosclerosis. They are discussed in detail. Mechanisms of fibrinolysis, in which plasmin or activators of plasmin occupy a key position, effect against this process. By histochemical estimation of this activator the fibrinolytical potential of the vascular system can be investigated under various clinical conditions. For this a series of instances is cited.


[Chemotherapy of gastrointestinal cancer (author's transl)]. Gastrointestinal cancer has proved exceedingly resistant to chemotherapy efforts. 5-Fluorouracil (5-FU) accepted as standard treatment, has provided only infrequent and incomplete antitumor effects. Other drugs as the nitrosoureas BCNU and CCNU or Mitomycin C do not match the effectiveness of 5-FU. Improvement in frequency of tumor regression have been recorded for gastric carcinoma with combinations of 5-FU and BCNU and 5-FU, adriamycin and Mitomycin C and for colorectal carcinoma with combination of 5-FU, methyl-CCNU and vincristine. There are also suggestions that such combination chemotherapy may produce increased survival when compared to untreated patients. The combination of 5-FU and streptozotocin in carcinoid tumors or adriamycin in primary hepatoma may be of some effectiveness.


Effect of immunosuppression on the growth of six murine tumors. Mice have been immunosuppressed with cyclophosphamide, cortisone-acetate, irradiation, or Ehrlich ascitic fluid (EAF) and then grafted with Ehrlich tumor or with one of the following strain-specific tumors: thymoma, methylcholanthrene-induced fibrosarcoma, B-16 melanoma, lymphatic leukaemia, and myeloid leukaemia. Immunosuppression of the host influenced very differently the growth of transplanted malignancies. The growth of thymoma and of Ehrlich tumor was regularly enhanced. The growth of fibrosarcoma and of melanoma, on the other hand, was retarded in mice pretreated with EAF and X-rays, or remained unchanged in mice pretreated with drugs. Leukaemia growth was not influenced by any immunosuppressive treatment; the only exception was enhanced growth of lymphoid leukaemia in animals pretreated with EAF. Thus different tumors grew differently in animals immunosuppressed by the same immunosuppressive agent, while different immunosuppressive treatment changed the growth of one particular tumor always in the same way. From this we concluded: (1) there is no rule as to how immunosuppression of the host will influence tumor growth; and (2) the way in which the malignant growth will be changed depends mainly upon the type of the tumor and probably not very much upon the type of immunosuppressive treatment.


Carcinogen-induced DNA repair in nucleotide-permeable Escherichia coli cells. Analysis of DNA repair induced by carcinogenic K-region epoxides and 1,2,3,4-diepoxybutane. Ether-permeabilized (nucleotide-permeable) Escherichia coli cells exhibited DNA excision repair when exposed to the following carcinogenic K-region epoxides: 7-methyl- and 7,12-dimethyl-benz[a]anthracene-5,6-oxide, chrysene-5,6-oxide and benzo[a]pyrene-4,5-oxide. This DNA excision repair was missing in uvr A and uvr B mutant cells. The K-region epoxide phenanthrene-9,10-oxide was ineffective in all E. coli strains tested. In contrast to the K-region epoxides which where found active only in wild type cells, 1,2,3,4-diepoxybutane and the 6,7-epoxides of the tumor promoter TPA (12-O-tetradecanoyl-phorbol-13-acetate) elicited DNA repair in uvrA, uvrB mutant cells as well. Enzymic activities catalyzing particular repair steps were identified by determining a) repair polymerization and b) size reduction of denatured DNA. A) An easily quantifiable effect in E. coli wild type cells was epoxide-induced repair polymerization. None of the K-region epoxides tested stimulated DNA repair synthesis in uvrA, uvrB mutant cells, indicating that the uvrA-, uvrB-controlled UV-endonuclease initiated excision repair by cleaving epoxide-damaged DNA. 1,2,3,4-Diepoxybutane and the TPA-6,7-oxides induced DNA repair polymerization in uvr-deficient cells, although to a lesser extent than in wild type cells, suggesting the involvement of uvr-independent incision steps. None of the epoxides induced repair polymerization in a mutant (polA107) lacking the 5'--3'exonucleolytic activity of DNA polymerase I (exonuclease VI). The absence of any repair polymerization in the polA107 mutant indicates that the exonuclease VI plays a central role in removing epoxide-damaged nucleotides. As evidenced by greatly reduced levels of repair polymerization measured in polA1 cells, DNA polymerase I was the main polymerizing enzyme. b) As a consequence of treatment with 7-methyl-benz[a]anthracene-5,6-oxide, DNA from wild type cells, contrary to uvrA mutant cells, showed size reduction after denaturation and sedimentation in alkaline sucrose gradients. This is explained by repair-specific endonucleolytic cleavage of damaged DNA. The incision required the presence of ATP indicating that functional UV-endonuclease needs ATP as a cofactor.


A note on the use of nitrate fertilizers in a high-risk geographical area for stomach cancer. Chile being the only country in the world producing natural fertilizers (NaNO3 and KNO3), it uses very large quantities of them for agricultural purposes. Since in early 1970 Zaldívar first related the use of fertilizers to stomach cancer mortality, it is of key interest to know the amount of nitrogen fertilizers used, expressed in tons of N, by province, for the period from 1945 to 69. Of a total of 420,750 tons of N for the nation, 238,950 tons were used in the high-risk provinces for stomach cancer (Colchagua, Curicó, Talca, Maule, Linares, Nuble, Concepción, Bio-Bio, Arauco, Malleco, Cautín). On the other hand, in the low-risk provinces (Tarapacá, Antofagasta, Atacama, Magallanes) only 2,550 tons were used. In Chile, the use of fertilizers increased from 37,550 (1945-50) to 185,050 (1965-69) tons of N.


The nucleotide-permeable Escherichia coli cell, a sensitive DNA repair indicator for carcinogens, mutagens, and antitumor agents binding covalently to DNA. Ether-permeabilized (nucleotide-permeable) Escherichia coli cells respond to alkylating and arylalkylating carcinogens with DNA excision repair, as assessed by their stimulation of DNA repair synthesis. In the present work, we have investigated whether DNA repair synthesis in ether-treated E. coli cells can serve as a general indicator to monitor the DNA-binding of carcinogens, mutagens and antitumor agents. Therefore, a standard assay was developed and comparative analyses were performed on 11 ultimate carcinogens, 10 proximate carcinogens, 2 tumor promoters, 6 mutagens, and 12 antitumor agents. All ultimate carcinogens (alkylating, acylating, arylalkylating agents) and mutagens (e.g., hydrogeen peroxide, acridine derivatives) caused DNA excision repair in wild type cells as measured by [3H] dTMP incorporation and simultaneously inhibited replicative DNA synthesis to various extents. Control experiments with the mutant cells uvrA and uvrB were performed to determine whether the pyrimidine-dimer-specific UV-endonuclease was involved in the removal of DNA damage. This was found to be true for the ultimate carcinogens (Ac)2 ONFln, mitomycin C, and for very reactive alkylating carcinogens. None of the ultimate carcinogens induced repair polymerization in mutant cells lacking the 5'-3' exonucleolytic activity of DNA polymerase I. Proximate carcinogens, such as Me2NNO, 4-nitroquinoline-1-oxide and aflatoxins, did not induce excision repair in the standard assay, probably because of the inability of E. coli to perform the activation steps necessary for covalent DNA-binding. However, Me2NNO, when pretreated with Udenfriend's hydroxylating mixture, gave rise to a low level of repair polymerization in ether-treated cells. Intercalating mutagens, such as quinacrine and ethidum bromide, inhibited replicative DNA synthesis. However, they were not found to be repair-inducers. THE TUMOR PROMOters TPA and phorbol-12,13-didecanoate did not cause excision repair, even when applied at high concentrations, nor did they inhibit repair synthesis stimulated by MeNOUr or (Ac)2 ONFln. The antitumor agents may be classified into two groups on the basis of the influence they exert on DNA synthesis: members of the first group (involving BCNU and bleomycin) stimulate repair polymerization and, in addition, inhibit DNA replication. These compounds are known to bind covalently to DNA. The second group of drugs (including adriamycin and cis-Pt(II)diammine complexes) inhibits DNA replication without stimulating repair synthesis. The predominant DNA-interaction of these compounds is known to be a non-covalent (i.e., intercalative, electrostatic) binding. Our experiments show that the ether-permeabilized E. coli cell can be successfully used to test ultimate carcinogens, mutagens and antitumor agents for repair-inducing and replication-inhibiting activity. The standard test might be extended to pre- and proximate carcinogens, provided these can be suitably activated.


Carcinogenicity of 4-chloronitrosopiperidine in Sprague-Dawley rats. 4-Chloronitrosopiperidine was administered to a group of 15 male Sprague-Dawley rats as a 0.005% solution in drinking water for 27 weeks. Of the 15 animals, 12 died with tumors induced by the treatment, half of them by the 41 st week; all 15 were dead at 58 weeks. Seven rats had liver tumors, 5 had esophageal tumors, 2 had tumors of the nonglandular stomach and 2 had tumors of the nasal turbinates. Several rats had tumors of more than one organ.


Determination of thallium in brain tissue by stable isotope dilution and field desorption mass spectrometry. The utility of field desorption mass spectrometry for quantitative metal cation analysis in forensic sciences is demonstrated by the determination of a lethal thallium level in the brain tissue of an experimental animal. Stable isotope dilution and accumulation of the electrically recorded field desorption ion currents with a multi-channel analyzer allowed a direct estimation of thallium in homogenized tissue samples without further pretreatment. Experiments with standard solutions revealed the limit of detection for thallium to be about 10 pg of the metal cation.


Mössbauer spectroscopy on oxygenated sperm whale myoglobin: evidence for an Fe3+-O2- coupling at the active center. 57Fe Mössbauer spectra of oxygenated sperm whale myoglobin (MbO2) show a well resolved quadrupole doublet with a temperature dependent splitting. The temperature dependence of the corresponding electric field gradient tensor (EFG) can be calculated from a Fe3+ term scheme for the iron at the active center. The Mössbauer spectra as well as the diamagnetc character of the MbO2-complex are then understood by an exchange coupling of the Fe3+-ion with O2- oxygen molecule ion. The resulting groundstate is a diamagnetic singlet. In order to keep the whole complex diamagnetic at room temperature, an exchange coupling with [J] greater than or equal to 300cm-1 is necessary. As the whole model is in fair agreement with many other spectroscopic data, it is believed to be a good starting point for further detailed calculations.


Is oxygen consumption of surviving tissues determined by the oxygen tension of the suspension medium? Oxygen tension in the suspension medium and oxygen uptake of the isolated diaphragm of the mouse have been determined simultaneously with systematic variation of shaking frequency and oxygen concentration of the gas phase. Oxygen tension in the suspension medium reaches its final value during the usual equilibration period of 10 minutes and remains constant for the whole measuring period. Each combination of shaking frequency and oxygen concentration of the gas phase results in a reproducible value of oxygen tension in the medium. Equal oxygen tension in the fluid phase can be attained by different combinations of shaking frequency and oxygen concentration of the gas phase. Oxygen uptake at the same oxygen tension of the suspension medium can be very different; it is the higher the higher the shaking rate in the combination. On the other hand the same oxygen consumption of the tissue samples can be observed with highly different oxygen tensions in the medium. It must be concluded from the data in this paper, that the influence of shaking rate per se on the oxygen uptake of isolated tissue can be of the same order as the influence of oxygen tension.


Conformationally restricted adenine nucleotide analogs in mitochondrial adenine nucleotide transport. The conformationally restricted adenine nucleotide analogs 8,3'anhydro-8-oxy-9-(beta-D-xylofuranosyl)adenine-5'-O-tri(di)-phosphate (I), and 8,2'-anhydro-8-oxy-9-(beta-D-arabinofuranosyl) adenine-5'-O-tri(di)-phosphate (II), were prepared chemically as their alpha-32P-labelled compounds and compared with syn-structured 8-bromo AT(D)P in mitochondrial adenine nucleotide translocation. The experimental findings demonstrate that the heterocycle-ribose orientation affects the carrier mediated adenine nucleotide transport very strongly, i.e. a non fixed adenine heterocycle in the anti region is prerequisite for the bound nucleotide to induce the transfer action of the adenine nucleotide carrier.


Alterations in ecdysone content during the post-embryonic development of Chironomus thummi: correlations with chromosomal puffing. The ecdysone titer of larvae and pupae of Chironomus thummi was determined by radioimmune assay (RIA) and revealed a concentration of about 150 ng/g fresh weigth in late third instar larvae and a peak of more than 450 ng/g fresh weight just preceding pupation. The ecdysone titer curve shows a high degree of correlation with previously observed puffing activity at the ecdysone sensitive chromosomal site IIIdl. Further, beta-ecdysone is the only endogenous ecdysone detected by high pressure liquid chromatography during larval-pupal development. It was also observed that developmentally arrested Chironomus contain less ecdysone than "normal" larvae.