[Primary liver cell carcinoma, aetiology and clinical picture (author's transl)]. Geographic area, age and sex influence the epidemiology of hepatoma. Aetiological factors are aflatoxins, sex hormones, thorotrast, alpha 1-antitrypsin deficiency, immunosuppression, vinylchloride, parasites, cirrhosis of the liver, and the hepatitis-B virus. Early diagnosis of the tumour is possible using alpha 1-fetoprotein estimations and modern morphological methods, particularly scintiscanning. Tumour resection is therapeutically desirable, while selective chemotherapy remains palliative and liver transplantation failed to prolong survival.


Suppression of experimental allergic neuritis with P2 protein of PNS myelin. Experimental allergic neuritis was induced in the guinea pig with bovine PNS myelin. Treatment with P2 protein at the onset of the disease markedly reduced the severity of the clinical signs compared with untreated controls and animals treated with CNS myelin basic protein. The mortality was reduced from 94% in the combined control group to 50% in the treated group. Saline was a better vehicle for administration of the suppressive inoculation than incomplete Freund's adjuvant.


[Population genetic examination of esterase D in the Lübeck area (author's transl)]. 1251 blood samples of non-related human beings coming from the area of Lübeck have been examined. Frequencies of the esterase D allel were EsD1 = 0.9001 and EsD2 = 0.0999.


[The detection of gamma globulin and Inv-factors in semen and saliva and of haptoglobins in semen (author's transl)]. Gm- 1-, 2-, and Inv 1-factors can be demonstrated in semen and saliva. For the examination of traces it is necessary to verify the suitable dilutions for the different charges of antisera and to test the eluates of the samples if they have a sufficient concentration. We observed some incorrect negative results in seminal and saliva stains which were apparently caused by insufficient material. The demonstration was independent of the secretor type. Haptoglobin could not be determined in semen and saliva.


Adult respiratory distress syndrome. A histopathologic study. The results of an anatomohistopathologic study carried out on subjects who died from various causes in resuscitation centers are reported with specific reference to the evolutive phases of the adult respiratory distress syncrome (ARDS). While a precise anatomohistopathologic diagnosis of non-clinically diagnosed ARDS in its initial phase is considered possible, the importance of recognizing this pathology in cadavers and living patients is emphasized because of the medico-legal implications involved.


[Results of experimental animal studies on pervascular influences of indwelling intravasal catheters upon free tissue grafts (author's transl)]. Free tissue grafting procedures were performed on sixteen experimental animals on the basis of previously described experiments (first communication). The objective of finding a suitable operating method for pervascular perfusion of epidermal grafts in animal experiments has been accomplished. At the same time, several possible ways of objectifying the results of blood gas analyses, temperature measurements, colorimetric determinations, and histological studies were subjected to tests.


[Possibility of using the BI-3 jet injector for gamma globulin administration]. The results of weighing glass flasks before and after "shooting" preparations varying in viscosity into these flasks from type BI-3 jet injectors indicated that the ejected dose of gamma globulin corresponded to the preset value. Studies carried out by means of radioisotyope technique with the use of Tc99m showed that the intramuscular injection of gamma globulin with type BI-3 jet injector resulted in 75% of the preset dose penetrating the body. For this reason in mass vaccination against Botkin's disease the dose of the preparation in the injector should be increased by 1/3.


[Erythrocyte diagnostica made from salmonellal phagolysates]. For the first time O antigens obtained from phagolysates were proved to be suitable for use as material for the production of highly specific erythrocyte diagnostic preparations. O antigens obtained from Salmonella by two methods, i.e. phage disintegration and Grasset's method, were subjected to comparative chemical analysis and found to have no essential difference. Nevertheless, the sensitizing potency of O antigens obtained from phagolysates were experimentally shown to be 3 times greater than that of O antigens obtained by Grasset's method. The optimum sensitizing doses established in the passive hemagglutination test for O antigens obtained by both methods indicated that these antigens were highly sensitive and specific.


[Study of the subclasses of immunoglobulin G. II. Qualitative and quantitative characteristics of IgG subclasses using antisera systems]. Bispecific antisera, or "antisera-systems", containing class- and subclass-specific antibodies to IgG were obtained from rabbits, goats and guinea pigs after brief courses of immunization with purified G1, G2, G3 and G4 paraproteins. After the elimination of antibodies to light chains by adsorption these antisera were tested in immunoelectrophoresis and radial immunodiffusion in gel with sera containing G paraproteins of different subclasses. In immunoelectrophoresis double lines and in radial immunodiffusion with G paraproteins of heterologous subclasses double rings were obtained: the external lines (or the external rings) were formed as a result of interaction between G paraproteins and antibodies to class-specific IgG determinants, the inner lines (or the inner rings) were formed as a result of interaction between the corresponding subclass of normal IgG and subclass-specific antibodies. The identification of different G paraprotein subclasses gave similar results when carried out with "antisera-systems" and with monospecific antisera to the corresponding IgG subclasses. "Antisera-systems" proved to be suitable for use in the identification of G paraprotein subclasses, as well as in the quantitation of different subclasses in normal IgG.


[Immunologic study of the cellular components of bacillus pyocyaneus. V. Antigenic analysis of slime and its fractions]. Various slime fractions were obtained from newly isolated mucoid strains of P. aeruginosa by the method of ultrafiltration or differential centrifugation with subsequent gel chromatography. Purified slime was found to react with a broader spectrum of typing O sera than the corresponding cell wall lipopolysaccharides. Erythrocytic diagnostic preparations produced on the basis of slime antigens allowed to reveale the presence of circulating antibodies against P. aeruginosa. The slime components with molecular weight of 30,000--100,000 daltons and greater contained common antigenic determinants, and the slime components with molecular weight of 10,000--30,000 daltons contained both specific antigenic determinants and those also common to the high molecular components.


[Therapeutic and preventive vaccination of persons bitten by wolves in the Aktiubinsk region]. A total of 46 cases of wolves attacking people were registered in the Aktyubinsk region during the period of 1972--1976. These attacks resulted in 1 person dying of wounds and 2 persons being infected with rabies. In more than a half of the wolves available for examination after the attack rabies was confirmed by laboratory investigation. All the victims of such attacks received antirabies gamma globulin (0.25--0.5 ml per kg body weight) and the maximum doses of rabies vaccine (daily injections of 5 ml for 25 days, then 2 more injections in the same dose 10 and 20 days later). No complications were observed after the injections. The vaccination schedule recommended for use in the USSR for the prophylaxis of rabies proved to be effective.


Immunoassay of macrophage stimulating alpha 2 SB (surface binding) glycoprotein in pig plasma following missile trauma. Pigs were subjected to a standardized missile trauma in the thigh and plasma samples were collected at various times after trauma. The plasma level of an opsonic alpha 2 SB (surface binding) glycoprotein related to macrophage phagocytosis was determined by an agglutination assay using 125I-labelled gelatin particles. The results indicated a transient but significance decrease (p less than 0.01) in the plasma agglutination activity after trauma reflecting a deficiency in plasma alpha 2 SB glycoprotein. One non-surviving animal showed a persistent decrease in the plasma level of alpha 2 SB glycoprotein, while the surviving animals showed a normalization indicating that this agglutination assay may have prognostic value during the course of posttraumatic therapy.


Serum alpha-fetoprotein assay in normal pregnant women. Perspectives of clinical use for purposes of diagnosis and prognosis. Radioimmunological assay of alpha-fetoprotein (AFP) has been performed in the serum of normal women at various stages of pregnancy and at the time of parturition; a total of 208 assay was carried out, AFP values rose until the 33rd week and at the time of childbirth dropped to levels about half of the maximum reached. No correlations were found between AFP values, neonatal weight and parity while the sex of the fetus appearech to have some influence. It is to be hoped that the application of these potential uses can be envisaged in the near future.


Acute hemorrhagic pancreatic necrosis in mice: lack of a pathogenetic role for complement. Feeding a choline-deficient diet containing 0.5% DL-ethionine induces an acute hemorrhagic pancreatitis in 100% of young female mice. Evidence for deposition of the third component of complement (C3) on acinar cell plasma membranes was sought, during the inductive stages, by a sandwich immunofluorescence technique. Such a localization could not be demonstrated even though the method is capable of detecting less than 8 x 10(-5) microgram of protein/mm2 of cell membrane. Artifactual binding of immunoglobulin reagents was encountered when goat antisera, with high levels of circulating immune complexes, formed the middle layer in the sandwich technique. This was attributed to the appearance of Fc receptors on the plasma membrane of degenerating acinar cells, and could be avoided by ultracentrifuging acinar cells, and could be avoided by ultracentrifuging the goat antisera prior to sue. In view of the fact that C3 cleavage represents an amplification loop in both the calssical and alternate pathways of complement activation, the lack of demonstrable C3 staining in tbe present experiments strongly suggests that complement plays no role in acinar cell necrosis in this model of pancreatitis.


CPC-PTA section staining of acid glycans. A cetylpyridinium chloride-phosphotungstic acid procedure suggested by Kelényi and Kiss (1976) for the staining of mast cell granules in ultrathin sections was slightly modified with regard to pretreatment, pH of incubation, and rinsing of sections. The method also proved suitable for the demonstration of various mucosubstances and of cartilage proteoglycan aggrebates.


[A ruthenium red-osmium tetroxide section staining method (author's transl)]. We suggest a method for the section staining of polyanions by means of ruthenium red and osmium tetroxide. A 5 min incubation in diluted ruthenium red-OsO4 solution pH = 7.4 of ultrathin Durcupan-sections previously treated with sodium methoxide results in the opaque stining of mast cell granules, cartilage proteoglycans, and various epithelial mucosubstances.


Decay of murine intestinal epithelial cells under extrusion. Electron microscopic findings of this study revealed 2 types of extrusion of brush border cells from the summits of intestinal villi: a) By expansion of initial vacuolation the degenerating cell enlarges and elevates itself above the level of the extrusion zone. Cells about to lift become sphered and are shed into the lumen for degradation to continue. b) Another group of cells vacuolating to a similar extend already may start apical decay in situ. In this case mechanical elevation, apparently, is inefficient or retarded such as to offer a cell the opportunity to decompose at its proper place in the mucosal lining. Contrary to type A extrusion, i.e. the desquamation of the entire degenerating cell, type B produces cellular debris to be shed into the lumen. Either type of exfoliation may be associated with the formation of gaps the closure of which proceeds slowly. The light microscopic test of cell viability by means of horseradish peroxidase demonstrated many brush border cells leaky such as to become loaded with the tracer irrespective of its vascular or enteric approach to the extrusion zone. Damage of the cell membrane due to a brief pretreatment with digitonin resulted in a dramatic rise of horseradish peroxidase positive cells. With respect to these findings, one cannot escape the conclusion that both types of extrusion confer intermittent leakage on the summits of intestinal villi which at least could account for part of the diffusion of macromolecules in either direction.


ATPase activity in the breast: a comparison between three methods. Adenosine triphosphatase enzymatic activity was investigated in human approximatively normal, dysplastic and neoplastic mammary tissue, by three different methods. Staining intensity varied within wide limits; myoepithelial cells and blood vessels showed similar enzymatic activity. Epithelial cells reacted only faintly, or not at all; carcinoma cells were never labelled. Stromal response was highly variable. The calcium-cobalt method of Padykula and Herman gave more intense reactions than the lead-nitrate procedure of Wachstein and Meisel, either in the original form or according to the modifications recommended by Russo and Wells. With the latter method the sharpness of stain deposits on the different structures was markedly enhanced. The functional significance of ATPase activity is discussed.


Uptake of 3H-testosterone in the skin of healthy women and in the skin of patients with acne vulgaris. Skin samples of interscapular region were taken from healthy women and patients with Acne vulgaris. The material was incubated with 3H-testosterone. The incubation of a part of the material was preceeded by an incubation with the unlabelled testosterone. An autoradiography was employed to detect and localize an uptake of 3H-testosterone in the skin. A significantly greater 3H-testosterone uptake was noted in the stratum basale and stratum spinosum of epidermis, sebaceous glands and hair sheaths and, in patients with Acne vulgaris, also within hair sacs and comedons.


Rapid changes in the serum total protein and globulin levels in complications caused by facultatively pathogenic Gram-negative bacteria. The changes in the levels of total protein and four globulin fractions were followed up throughout the entire course of complications caused by Gram-negative facultative pathogens in 37 acute cases of respiratory insufficiency accompanying different underlying illnesses and in 9 chronic, bedridden patients given artificial ventilation. At the onset of the infectious complications, in the first place in septic shock, the levels of various globulin fractions showed a decrease corresponding to a half-life of 2 to 4 days. Neither the increased catabolism, nor the protein losses by the urine and tracheal secretions offer a sufficient explanation for the escape of globulins of this extent from the plasma. It seems that this is a consequence of the increase in capillary permeability due to the effect of antigen-antibody reactions and that of endotoxin. As a result, in the critical phase of the infectious complications, at the point of culmination, e.g. in septic shock, diminished amount of different globulins is transported to the site of utilization, that is, to the inflammatory area.


Total protein, electrophoretic patterns, total amino acid and creatinine in the amniotic fluid of premature infants. Total protein, total amino acid, creatinine and the 5 major protein fractions were determined in amniotic fluid samples from 42 normal and 12 premature neonates. Total protein, albumin, globulin, total amino acid, creatinine and A/G ratio were significantly lower in the 6 premature infants who developed the respiratory distress syndrome (RDS) than in the controls. The alpha 1, alpha 2 and gamma globulin levels were within the normal ranges. In the 4 premature stillborn babies, total protein and globulins were significantly higher than those of the controls, while the albumin, total amino acid and creatinine levels were normal. A/G ratio for this group was still lower than that for the former group and much lower than the control ratio. The remaining 2 premature alive infants showed a significant decrease in the alpha 1, alpha 2 globulins and creatinine levels. Slight increase in total amino acid, and significant increase in the beta globulin, while total protein, albumin, gamma globulin and A/G ratio were within the normal ranges.


[Circadian variations in the frequency of arrhythmia and quantitative analysis of rhythm disturbances]. In 29 patients with long term ECG monitoring the circadian rhythm of extrasystoles was investigated. A decrease of maximal frequency of extrasystoles was calculated for each hour and an increase of the minima during night was detected. In 77% of the patients also for 6 hour-segments a circadian rhythm could be calculated. The effect of the antiarrhythmic treatment on the frequency of extrasystoles was different depending on the frequency of the sinus rhythm.


[Diagnostic progress in testicular carcinoma]. The diagnostic value of the serum tumour markers, beta-HCG and AFP, is investigated in 71 patients with malignant testicular tumours. Regular screening with beta-HCG and AFP, has improved the early diagnosis of recurrences and metastases in nonseminomatous testicular tumours.


Lymphocyte transformation in the presence of myelin basic protein in multiple sclerosis and control subjects. The transformation of lymphocytes in vitro in the presence of human myelin basic protein has been investigated in normal people, patients with multiple sclerosis (MS) and controls with other neurological diseases. There was little or no response at low concentrations (1--10 microgram/ml) but significant transformation at higher concentrations (100--1000 microgram/ml) in all three groups. There was no significant difference among the groups as a whole, but those MS patients who had had disease for more than 10 years did show greater responses than normal subjects (P less than 0.05). Increased responses could not be correlated with any other aspect of disease activity: in particular they were not increased in patients with acute relapses. The use of autologous serum instead of homologous AB Rhesus positive serum did not significantly alter lymphocyte responsiveness. The absence of any response in the presence of purified calf thymus histone suggests that the response to myelin basic protein indicates a low level of lymphocyte sensitization to this antigen even in normal subjects. The present evidence does not support a primary pathogenetic role for such a reaction in MS. The increased response in patients with a long duration of disease might merely be an effect of white matter damage or might represent an amplification of the normal immune response contributing to myelin breakdown and leading to the emergence of the progressive stage of the disease. The study of lymphocyte responsiveness over a wide range of concentrations of myelin basic protein is considered to resolve some of the controversy surrounding this subject in the literature.


Experimental subperiosteal implantation of porous Al2O3 ceramic for mandibular ridge augmentation. Porous Al2O3 ceramic material with pore size of 100-750 micrometers was used for implants placed subperiosteally as alveolar ridge augmentation onlays in six Cercopithecus monkeys. Clinical and radiographic examination during the postoperative period revealed rapid healing and anchorage of the implant in five of the animals. One implant was lost. The implants were retrieved after 4, 10, 11 and 12 months' implantation time. Histologic and microradiographic examination demonstrated fibrous and bone tissue invasion into the pore system at the base of the subperiosteal implant, with good anchorage of the ceramic material to the jaw. Only fibrous tissue was found to project into the ceramic openings on the implant sides facing the periosteum. Labeling of the mineralizing tissue was attempted using intravenously administered tetracycline (Reverin) and calcein (Calcein) at varying intervals prior to sacrifice, but the labeling results were not impressive.


The role of HLA-B27 in the diagnosis of low back pain. The HLA-B27 antigen was determined in 652 patients with low back pain which had lasted for more than 3 months. A clinical and roentgenological examination of the sacroiliac joints and the thoraco-lumbar spine was performed in all the patients. The control group consisted of 302 unrelated persons who did not show signs of low back pain. Antigen HLA-B27 was found in 276 of these 652 patients attending the ward for rheumatic diseases (42.4 per cent) and in 37 of the 302 unrelated persons in the control group (12.2 per cent). The difference is statistically highly significant (P less than 0.001). Ankylosing spondylitis was found in 128 out of the 276 patients with low back pain and antigen HLA-B27. This demonstrates the importance of this antigen in the differential diagnosis of low back pain.


Immunohistochemical demonstration of intracellular immunoglobulin in formalin fixed, paraffin embedded sections, as staining method in diagnostic work. 52 specimens from 34 patients with multiple myeloma and 3 cases with reactive plasmacytosis were stained by the 2-step labelled or 3-step unlabelled immunoperoxidase (IP) technique for demonstration of intracellular immunoglobulin in order to test the utility of these methods as staining methods in routine diagnostic work. The specificity of the methods was high as agreement between IP staining and serumanalysis for M-component was achieved in 27 out of 28 cases, where complete serumanalysis were available. Interpersonal variation in interpretation of the staining results was low. With optimal dilutions of antisera the evaluations of the examiners were concordant in 51 out of 52 specimens. By comparing the different tissue specimens the best result was obtained with iliac bone biopsies both concerning IP staining and morphological preservation. No difference was observed between results obtained by the 2-step labelled and 3-step unlabelled IP techniques.


Effect of sensitization on spontaneous and phosphatidylserine-induced histamine release and on cyclic AMP and GMP levels in isolated rat mast cells. Sprague Dawley rats were sensitized with 20 microgram or 100 mg egg albumin (using pertussis vaccine as adjuvant). Mast cells isolated from the former group of animals showed a higher degree of histamine release upon challenge in vitro with egg albumin than those from the latter group. Using the lower amount of antigen for immunization mast cells from Hooded Lister rats showed an even higher degree of histamine release induced by antigen. An increased antigen-induced histamine release was associated with an increased spontaneous and phosphatidylserine-induced histamine release. Histamine release induced by phosphatidylserine was found to be specific in so far as it was calcium dependent and theophylline-inhibited. The basal level of cyclic AMP in mast cells was significantly depressed by sensitization. There was a relationship between the cyclic AMP/cyclic GMP ratio and the degree of spontaneous, phosphatidylserine-induced and anaphylactic histamine release. The results suggest that sensitization induces an increased release of histamine not only to the specific antigenic stimulus but also to more unspecific stimuli. Concomitantly there is a fall in the cyclic AMP/cyclic GMP ratio. The relationship between these two phenomena is discussed.


Palliative embolization of arterial renal tumour supply. Results in 10 cases. Palliative occlusion of the arterial renal tumour supply was performed in 10 patients and the follow up is reported. Nine of the patients had no subsequent nephrectomy. Spongostan (99% gelatin) was used as the embolic material in 4 patients with the addition of steel coils in 2. Bucrylate was used in 6 cases. Six patients are alive with survival rates presently ranging from 3 to 24 months after embolization. Improvement of the survival time cannot be estimated but local symptoms such as hematuria and pain may be treated in those patients with renal tumours who are not considered for surgery.


The development of mesosomes of Staphylococcus aureus during autolysis. Mesosomes of Staphylococcus aureus were examined morphologically under the electron microscope. Three different methods of specimen preparation (thin section, freeze-fracture, and negative staining) were used to eliminate artifacts due to sample processing. Mesosomes were rarely seen in intact cells but were quite distinct in autolysed cells incubated in 1.2M sucrose -0.33M acetate buffer (pH 5.8) at 25 degrees C. The phospholipid content increased by 20% of the control as autolysis proceeded. Since the plasma membrane did not show any other significant changes, the development of mesosomes during autolysis was shown to be a real event. Most of the well-developed mesosomes were found at the septum in negatively stained specimens. Initial wall-lysis occurred at this site, suggesting a close relationship between autolysis and the development of mesosomes in S. aureus.


Purification of antilymphocyte antibody (ALA) from patients with systemic lupus erythematosus (SLE)-immunoabsorption and elution. Purification of antilymphocyte antibody (ALA) from patients with systemic lupus erythematosus (SLE) was achieved by immunoabsorption and elution. Human tonsil cells or thymocytes were used as absorbents. Complement dependent microcytotoxicity tests showed that, in comparison to the parent sera, the eluate from tonsil cells was eight times, and that from thymocytes four times, more active. Antinuclear activity was eliminated by elution. The ALA was almost entirely IgM, IgG being involved in only a few cases. IgA lacked cytotoxic activity. ALA was directed at both T- and B-cell surface determinants, which suggests that, in SLE, it has a heterogeneous biological composition.


Use of recombinant DNA technology for the production of polypeptides. DNA sequences corresponding to specific genes may be prepared by chemical synthesis, isolation of naturally occurring DNA, or reverse transcription. Such DNA may then be inserted into vectors such as plasmids or bacteriophages which carry the DNA into bacterial cells. Although significant differences exist in the basic molecular biology of eucaryotic and procaryotic organisms, these differences do not constitute absolute barriers to the expression of eucaryotic genes in bacteria. Several eucaryotic proteins, including insulin, growth hormone, ovalbumin, dihydrofolate reductase and somatostatin have been produced in bacteria. The use of chimeric microorganisms harboring recombinant DNA offers a completely new approach to the production of biologically useful polypeptides.


Detection and possible functional influence of antibodies directed againt the pancreatic islet cell surface. Antibodies directed towards determinants on the surface of rat islet cells can be detected qualitatively by the indirect immunofluorescence or quantitatively by a specific radioassay for IgG. Antibodies are found in insulin-dependent diabetics or in rabbits immunized with islet cells. Such antibodies may affect the B-cell function as indicated by the inhibition of incorporation of radioactive leucine into proinsulin/insulin in vitro.


Inhibition of histamine release and ionophore-induced calcium flux in rat mast cells by lidocaine and chlorpromazine. We studied the effects of lidocaine (L) and chlorromazine (C), two compounds known to affect the binding of calcium to cell membranes, on histamine release and calcium uptake by purified mast cells upon challenge with the ionophore A23,187 or with compound 48/80. At low concentrations L and C inhibited the Ca++ flux as well as histamine release while higher concentration caused enhancement in this function. Evidence was obtained that L 10(-4) M may displace Ca++ from the cell membranes.


Mimickry of anti-granuloma effect of prostaglandin E by dibutyryl cyclic-AMP and some phosphodiesterase inhibitors. Local administration of PGE1 to pre-formed inflammatory granulomata of rats results in a decrease of granulomatous tissue and reduction of prostaglandin concentrations in granulomatous exudates. Under the same experimental conditions, a similar correlation between these two effects is observed with dibutyryl cyclic-AMP. An anti-granuloma effect is also achieved with the phosphodiesterase inhibitors, IBMX and RA-233, but not with theophylline, a rather feeble inhibitor of this enzyme. The present findings provide further support for the concept that elevation of cyclic-AMP in cell population(s) within granuloma is a promising line for pharmacological suppression of inflammatory tissue proliferation.


Retrolental fibroplasia.

Retrolental fibroplasia. Retrolental fibroplasia is more accurately referred to as retinopathy of prematurity. Host factors most clearly incriminated are the maturity of the infant and the time in oxygen therapy. A continuum of retinopathy ranges from abnormal arborization of terminal retinal vessels to a detached retina incorporated in a cicatrix behind the lens. Every high-risk infant should be examined by indirect ophthalmoscopy prior to discharge from the nursery. A thorough knowledge of the natural course of the disease is essential because many cases will resolve spontaneously.


Evaluation of the chronic inhalation toxicity of a manganese oxide aerosol--I. Introduction, experimental design, and aerosol generation methods. A brief literature review on manganese toxicity is presented; as related to designing a chronic inhalation study for evaluating methylcyclopentadienyl manganese tricarbonyl when utilized as a motor fuel additive. The experimental design of this study is described. The generation system utilized to simulate the manganese aerosol produced by an internal combustion engine is described in detail. This generation system operated twenty-four hours per day, seven days per week producing aerosols at 11.6, 112.5, and 1152 micrograms Mn/m3 with an aerodynamic diameter of approximately 0.11 micron.


Histamine and cyclic AMP in isolated canine parietal cells. The relationship between cyclic AMP production and the response of isolated canine parietal cells to histamine has been examined. Histamine increased cyclic AMP generation, and this effect correlated with histamine stimulation of oxygen consumption and aminopyrine accumulation. Metiamide inhibited histamine-stimulated cyclic AMP generation and oxygen consumption in a parallel fashion. At concentrations below 100 microM, isobutyl AMP production and oxygen consumption in a similar fashion. However, with IMX above 100 microM, histamine caused no further increases in oxygen consumption, despite markedly enhanced cyclic AMP generation. Neither carbachol nor gastrin increased cyclic AMP production beyond that produced by IMX alone, and the combinations of histamine and carbachol and of histamine and gastrin produced no greater cyclic AMP generation than produced by histamine. These findings support a close relationship between cyclic AMP production and the action of histamine but not of carbachol or gastrin on isolated parietal cells. The mechanisms underlying the potentiating interactions between histamine, carbachol, and gastrin involve step(s) beyond stimulation of cyclic AMP generation.


Molecular forms of immunoreactive pancreatic cationic trypsin in pancreatitis patient sera. The molecular forms of immunoreactive pancreatic cationic trypsin in sera of patients with acute pancreatic inflammation have been characterized using a radioimmunoassay technique that is capable of detecting trypsinogen as well as trypsin bound to alpha 1-antitrypsin. Trypsin bound to alpha 2-macroglobulin is not immunoreactive under normal assay conditions. However, alpha 2-macroglobulin-bound trypsin can be detected after gel filtration of serum on Bio-Gel A-0.5 m and acid treatment of column fractions. The average serum level of immunoreactive cationic trypsin from 20 patients with acute pancreatic inflammation was 1,590 ng/ml. An average normal value of 26 ng/ml has been obtained previously. Serum samples from 14 patients with pancreatic inflammation were chromatographed under conditions that resolve trypsinogen, alpha 1-antitrypsin-bound trypsin, and alpha 2-macroglobulin-bound trypsin. In each case, the major portion of the immunoreactive material eluted at a position corresponding to free trypsinogen, while a minor fraction of the immunoreactive material appeared to be trypsin bound to alpha 1-antitrypsin. The zymogen nature of the major peak was confirmed in one case by activation with human enteropeptidase. In 11 of 14 patients, acid treatment of the alpha 2-macroglobulin peak yielded immunoreactive trypsin.


[Immunopathogenesis of the Vogt-Koyanagi-Harada syndrome]. Regarding immunopathogenesis of the Vogt-Koyanagi-Harada syndrome (VKHS), peripheral blood lymphocytes of a female patient with this disease were tested for sensitization against several antigens such as myelinic basic protein (BP) and uveoretinal, brain and spleen tissue homogenates. For comparison, cells of seven patients with chorioretinitis only and cells of six patients with encephalitis of unknown etiology were also tested. Cells of healthy donors served as controls. The electromobility test and the leucocyte migration test were used as in vitro test systems for cell-mediated immunological reactivity. The results show a strong sensitivity against BP and uveoretinal and brain tissue homogenates in VKHS. In the patients suffering from chorioretinitis, a strong reactivity against uveoretinal homogenate only was found, while in patients with encephalitis, responses to brain antigens only were observed. Reactions towards spleen tissue homogenate were negative in all experiments. The extent to which this tissue-specific immunological reactivity, which is different from the monoreactivity in chorioretinitis or encephalitis, might be a characteristic phenomenon of the VKHS is discussed.


Fluorescein angiography and changes in aqueous humor protein after argon laser photomydriasis in rabbits. In brown-eyed and blue-eyed rabbits, irises treated with argon laser photomydriasis (ALP) were studied by using fluorescein angiography and by evaluating the aqueous humor protein concentration. The following results were obtained. Immediately after ALP, a breakdown of the blood-aqueous barrier is demonstrated by a marked influx of fluorescein into the ciliary processes and into the anterior and posterior chambers. The dye flow into the aqueous humor decreases to normal values within 3 days. There is no fluorescein leakage from laser impacts themselves. Peripupillary capillary loops are permanently occluded, but in a few cases, recanalization occurs. Neovascularization did not occur during 56 days of observation. ALP initially leads to a six- to eightfold increase in normal aqueous humor protein concentration, which decays to normal values within 3 days.


Analysis of antigens in a commercial house-dust extract by means of quantitative immunoelectrophoresis. Crossed immunoelectrophoresis (CIE) of a commercial house-dust extract revealed 43 immunoprecipitates. Rocket immunoelectrophoresis with 106 different allergen extracts from animal hair and dander, feathers, fungi, pollen, foodstuffs, textiles and house-dust mites showed the presence of precipitating antibodies in the rabbit anti-house-dust serum against antigens from 34 of the extracts. Crossed immunoelectrophoresis with the addition of antigen, and crossed line immunoelectrophoresis (CLIE) with these 34 allergen extracts, as well as crossed immunoelectrophoresis with intermediate gel containing rabbit antibodies against nine human serum proteins, allowed the identification of 14 antigens in the house-dust extract. These antigens were shown to originate from house-dust mites, hair and dander from various animals, fungi, feathers, human hair and dander, and human serum proteins.


Specific diagnosis of exogenous bronchial asthma in adults. Comparison between case history, intracutaneous test, RAST, histamine release from basophil leucocytes, and bronchial provocation. Twenty-two adults with extrinsic bronchial asthma were examined with 26 different allergens from five groups: house dust + mite (30 BP), animal dandruff (30 BP), pollen (12 BP), mould fungi (11 BP), and feather + cotton (10 BP). The same allergen batch was used for each allergen extract in all analyses. Based on the results of the single analyses: case history (CH), intracutaneous test (ST), RAST, histamine release from basophil leucocytes (HR), and the results of the analysis combinations and the "true diagnosis" bronchial provocation (BP), the sensitivity and specificity (SE and SP) were calculated and shown graphically. An evaluation of the diagnostic reliability of the single analyses and analysis combination was then possible. Generally, HR was the best individual analysis with very few false positive values and false negative values, followed by RAST, ST, and CH. None of them was able to predict allergy for feather + cotton. Otherwise, BP was positive when HR and rast reached scores 2 and 3 with the exception of RAST for animal dandruff. As regards ST, we had to reach score 3 in order to predict positive BP. CH would only provide the diagnosis with a maximum score in the pollen group. BP was positive in any analysis combination resulting in score 6 or more. ST appeared to be the most sensitive of the individual analyses. All the patients with ST score 0 showed negative BP for all allergen groups, whereas a corresponding sensitivity for HR score 0 was only achieved with animal dandruff, and for RAST score 0 for pollen only. In spite of the new analyses, BP is in many cases necessary if a definite diagnosis is required.


Ultrastructural localization of histones in epiphyseal chondrocytes. Epiphyseal growth plate chondrocyte nuclei of growing rats were studied for histone content using the ammoniacal silver technique. This stain, which is specific for the demonstration of the arginine-rich fractions of histones, revealed that growth plate nuclei vary in their histone content. Nuclei of cells of the proliferating zone revealed a significantly greater amount of postformalin ammoniacal silver deposit consistent with the presence of arginine-rich histones.


Malignant premature ventricular beats in ambulatory patients. The characteristics of premature ventricular beats predisposing to ventricular tachycardia or fibrillation were assessed by 24-h ambulatory monitoring and maximal treadmill exercise testing in 339 cardiac patients with premature ventricular beats. Premature ventricular beats were divided into early (Q-premature ventricular beat less than QT), late (within the last 20% of the cardiac cycle), and midcycle. Ventricular tachycardia was recorded in 45 patients and ventricular fibrillation, in three. The frequency of ventricular tachycardia or fibrillation was 32% in patients with late, 16% in patients with early, and 7% in patients with midcycle premature ventricular beats (P less than 0.05). Patients with frequent (less than 10/min) multiformed premature ventricular beats had a frequency of ventricular tachycardia or fibrillation of 44%, while only 13% of patients with frequent uniformed premature ventricular beats had ventricular tachycardia (P less than 0.05). Ambulatory patients with ventricular tachycardia or fibrillation have frequent multiformed premature ventricular beats, and the ventricular tachycardia or fibrillation is usually triggered by late premature ventricular beats.


Presence of fibronectin in basement membranes and acidic structural glycoproteins from human placenta and lung. Cold-insoluble globulin was detected in both trophoblast and alveolar basement membrane preparations, whereas it was not detected in GBM preparations. Acidic structural glycoproteins from both placental villi and lung parenchyma, which were extracted with 0.3 M acetic acid and recovered by adjusting the pH to 4.7, also contained CIg. Fractions of TBM, solubilized by either dilute alkali (0.01 N NaOH), by reduction and alkylation of the disulfide bonds, or by 0.3 M acetic acid extraction, all contained the antigen and possessed properties similar to those of ASG. The ASG fractions also reacted with antifibrinogen, but proof that the two types of determinants occur on a given molecular species is lacking at present. Purified collagenase solubilized CIg from ABM, from lung parenchyma, and from the stroma of placental villi, and this finding is strong evidence for an association of CIg with collagen in these connective tissues.


Transabdominal bilateral adrenalectomy for metastatic breast carcinoma. A case is made for the more general adoption of the transabdominal approach to the adrenal glands as this has obvious advantages over bilateral loin incisions.


Studies on filariasis transmission in Kwale, a Tanzanian coastal village, and the results of mosquito control measures. The main vectors of bancroftian filariasis in Kwale, Tanzania, were Anopheles gambiae complex and Culex p. quinquefasciatus. After vector control by the insecticides Dursban (chlorpyrifos) or Abate (temephos), the number of all man-biting mosquitoes was reduced by 94.8%, and that of house-resting mosquitoes by 95.3%. Anopheles gambiae complex were highly exophilic, possibly due to a high proportion of Anopheles merus in this village. The infection rate of A. gambiae complex rose significantly in the post-control samples (P less than 0.05), but nevertheless the overall reduction in filariasis transmission was 87.2%. The infection rate of A. gambiae complex caught off human bait was significantly higher than that of the spray-caught sample (P less than 0.05). It was suggested that the former sample contained a higher proportion of A. merus. This species may be a better vector of filariasis than freshwater forms.


[Action of isoprinosine on the "in vitro" activation of human lymphocytes (author's transl)]. The action of isoprinosine on lymphocytes stimulated by optimal or suboptimal concentrations of polyclonal mitogens (ConA and PWM) was studied in 3, 5 and 7-days cultures. In some experiments, varying concentrations of isoprinosine were added to lymphocyte cultures at variuos times. Isoprinosine exerted no effect on non stimulated cells but enhanced the proliferative response in the presence of mitogens. The optimal response was obtained with a dose of 1 000 micrograms (/10(6) cells) of isoprinosine and on the 5th day in the cultures stimulated by ConA or PWM, but an enhancing effect could still be detected on the 7th day of culture. The effect seems stronger when isoprinosine is introduced earlier in the course of the cultures. Preliminary experiments suggest that isoprinosine increase the proliferation of both T and B cells and that its activity on B cells could be due to the potentiating action of helper T cells.


[A quick and inexpensive staining method for vaginal smears]. Vaginal cytology is a commonly used method for diagnosis of sexual cycle phases in the rat, but all staining techniques are time consuming. A simple, easy and fast technique is reported here, that is based upon the use of a Lugol solution that allows precise identification of the sexual cycle day of the rat. Following this technique, samples are taken at 8:00 a.m. and a smear is made on the glass slide adding one drop of Lugol solution. This preparation is covered and seen directly under the optic microscope.


[Ascending aortic aneurysm with mucoid infiltration of the media and aortic leaflets. Pathogenic hypotheses- surgical consequences]. The theoretical and practical problems posed by aneurysms of the ascending aorta with mucoid infiltration of the Media and the aortic leaflets are discussed with reference to two suggestive cases. These histological changes are not specific for Marfan's syndrome, the relation with which is very inconstant. Incomplete replacement of the affected tissues risks a recurrence which is especially serious when the sinus of Valsalva is involved. Therefore, total replacement of the ascending aorta with aortic valve replacement and reimplantation of the coronary arteries is generally indicated. Histology of the excised tissues should be performed routinely with specific stains for mucopolysaccharides. Close follow-up of operated patients with this condition is recommended, especially those with only partial replacements.


[Value of continous 12-hour electrocardiography during the rehabilitation period in patients convalescing from myocardial infarction. Apropos of 40 cases]. The results of continuous 12-hour electrocardiography in 40 convalescent patients after myocardial infarction undergoing a programme of readaptation on average 4 weeks after the initial infarction are reported. The frequency and alarming nature of the VES observed in 75% patients who had already undergone selection for the coronary readaptation programme, 30% of whom were receiving antiarrhythmic therapy, is emphasized. The observation of a progressive increase in the number of VES during the day prompts the following suggestions: the possibility of limited continuous ECG monitoring at the end of the day, the need to adapt the dosage of antiarrhythmic drugs during this most critical period. The comparison of the results of continuous ECG with those of repeated exercise ECG seems very positive, but some reservations are made and it would seem logical to associate these two complementary methods of investigation.


[Anti-arrhythmic effect of magnesium cations. Protection against ventricular excitability during cardiotonic therapy]. This study showed the effectiveness of magnesium sulphate during digitalis therapy complicated by ventricular hyperexcitability. 28 patients with cardiac disease and ventricular arrhythmias in heart failure were studied. Magnesium sulphate was given by slow intravenous infusion (30--50 mg/min) twice daily. The anti-arrhytmic action allowed treatment with digitalis until improvement in cardiac function was observed. The mechanism of the action of magnesium sulphate in digitalis--induced arrhythmias is discussed.


Latex fetuin spheres as probes for influenza virus neuraminidase in productively and abortively infected cells. Fetuin bound latex spheres do not adhere to the membranes of non-infected cells but adhere to those of cells productively infected by fowl plague virus (FPV Dobson strain). In contrast, asialo fetuin spheres do not attach to the membranes of productively infected cells. Moreover latex fetuin spheres incubated with extracts of productively infected cells and extensively washed are specifically enriched in neuraminidase activity without any trace of haemagglutinin. These observations suggest that viral neuraminidase in the membrane is the site of attachment of the sialic acid moieties of fetuin spheres. These neuraminidase sites are detectable when L cells are productively infected by a mammalian cell adapted mutant of the Dobson strain (FPV-B) but are not detectable on L cells abortively infected by wild type (FPV+). However, even in the abortive system, neuraminidase is synthesised de novo as shown by its labelling with 14C-glucosamine and by its isolation from labelled extracts of infected cells by latex fetuin spheres. These results show that misintegration of viral neuraminidase in the plasma membrane of L cells is a feature of abortive infection of these cells by the Dobson strain of FPV. However the relationship (if any) of this misintegration to abortive infection remains to be established.


[Method of histological diagnosis of liver alveococcosis]. A method for histological diagnosis of alveococcosis of the liver by staining sections with aniline gentian stain is proposed which permits to detect clearly scraps of the cuticular membrane and pre-vesical formations of the parasite even in those cases when small amounts of biopsy (operation) materials are brought for examinations.


[Pulmonary cryptococcosis].

[Pulmonary cryptococcosis]. A clinico-morphological analysis of two observations of cryptococcomas of the lungs are presented. In one case the diagnosis was established on the basis of cytological examinations of the sputum. This form of pulmonary cryptococcosis as well as identification of cryptococci in the sputum by cytological examinations are first described in the Soviet literature. The attention is drawn to the difficulties of clinico-roentgenological and morphological diagnosis of this disease and to the necessity to differentiate it from metastases of hyperneophroma, diseases of parasitic origin, and mycoses by other fungi.


Ultrastructure of peripheral nerves of mice inoculated with rabies virus. Fourty adult female albino mice were inoculated in the right hind leg with rabies viruses of the street type. The mice were sacrificed with an interval of 24 hours each, starting in the next day after inoculation. From the 10th day ownwards the animals started presenting signs of paralysis, first on the leg where the viruses were inoculated anbnormalities were found in peripheral nerves compatible with axonal degeneration with secondary demyelination but the rabies viruses were not found in the axoplasm, myelin sheet, Schwann cell cytoplasm, endoneural or in the epineural structures.


[Familial spastic paraplegia with amyotrophy. Clinical, electromyographic, histochemical study and microdissection]. Four cases of familial spastic paraplegia with amyotrophy in siblings from a consanguineous married are reported. The routine laboratory examination were normal. The electromiography and muscle biopsy processed by histochemistry showed signs of denervation with reinervation. The motor nerve conduction velocity was decreased in the peroneal nerve in 3 cases. The teased fiber preparation of sural nerves was abnormal in four cases. It was found increased of C, D and G fibers suggesting demyelination with secondary remyelination. The authors believe the abnormalities found could be due the distal axonal degeneration, with secondary regeneration and suggest the hypothesis that the fact is an axoplasmic flow defect in the central and peripheral nervous system.


Inhibition of platelet aggregation during hypovolemic hypotension by acetylsalicylic acid and artificial respiration. In vivo measurement of platelet aggregation was performed by a previously described photoelectric method. It is shown that platelet aggregation induced by arterial hypotension can be inhibited by i.v. administration of ASA and by artificial respiration. The inhibition of platelet aggregation following artificial ventilation is significantly more pronounced than by ASA given 24 h prior to the beginning of the experiments. Different sources of platelet aggregation in venous and arterial blood are informative about the canine lung not only as a filter for aggregates but also as a source of aggregating substances.


In vivo microscopy of the hepatic microvascular system. In vivo microscopic study of the responses of the various segments of the hepatic microvascular system to adrenergic, cholinergic, and other aminergic substances and their antagonists have revealed the relative distribution of receptor sites in these various vessels. The results also suggest a possible interaction of cholinergic substances with periportal mast cells. The release of serotonin, histamine and possibly other products from these cells results in dramatic alterations in blood flow through the hepatic sinusoids due to the adhesion of leukocytes and platelets to the endothelium.


The electrophoretically 'slow' and 'fast' forms of the alpha 2-macroglobulin molecule. alpha 2-Macroglobulin (alpha 2M) was isolated from human plasma by a four-step procedure: poly(ethylene glyco) fractionation, gel chromatography, euglobulin precipitation and immunoadsorption. No contaminants were detected in the final preparations by electrophoresis or immunoprecipitation. The protein ran as a single slow band in gel electrophoresis, and was designated 'S-alpha 2M'. S-alpha 2M bound about 2 mol of trypsin/mol. Treatment of S-alpha 2M with a proteinase or ammonium salts produced a form of the molecule more mobile in electrophoresis, and lacking proteinase-binding activity (F-alpha 2M). The electrophoretic mobility of the F-alpha 2M resulting from reaction with NH4+ salts was identical with that of proteinase complexes. We attribute the change in electrophoretic mobility of the alpha 2M to a conformation change, but there was no evidence of a change in pI or Strokes radius. Electrophoresis of S-alpha 2M in the presence of sodium dodecylsulphate gave results consistent with the view that the alpha 2M molecule is a tetramer of identical subunits, assembled as a non-covalent pair of disulphide-linked dimers. Some of the subunits seemed to be 'nicked' into two-thires-length and one-third-length chains, however. This was not apparent with F-alpha 2M produced by ammonium salts. F-alpha 2M produced by trypsin showed two new bands attributable to cleavage of the subunit polypeptide chain near the middle. Immunoassays of F-alpha 2M gave 'rockets' 12-29% lower than those with S-alpha 2M. The nature of the interactions between subunits in S-alpha 2M and F-alpha 2M was investigated by treating each form with glutaraldehyde before electrophoresis in the presence of sodium dodecyl sulphate. A much greater degree of cross-linking was observed with the F-alpha 2M, indicating that the subunits interact most closely in this form of the molecule. Exposure of S-alpha 2M to 3 M-urea or pH3 resulted in dissociation to the disulphide-bonded half-molecules; these did not show the proteinase-binding activity characteristic of the intact alpha 2M. F-alpha 2M was less easily dissociated than was S-alpha 2M. S-alpha 2M was readily dissociated to the quarter-subunits by mild reduction, with the formation of 3-4 new thiol groups per subunit. Inact reactive alpha 2M could then be regenerated in high yield by reoxidation of the subunits. F-alpha 2M formed by reaction with a proteinase or ammonium salts was not dissociated under the same conditions, although the interchain disulphide bonds were reduced. If the thiol groups of the quarter-subunits of S-alpha 2M were blocked by carboxymethylation, oxidative reassociation did not occur. Nevertheless treatment of these subunits with methylammonium salts or a proteinase caused the reassembly of half-molecules and intact (F-) tetramers. It is emphasized that F-alpha 2M does not have the properties of a denatured form of the protein...


Sulphate-ion/sodium-ion co-transport by brush-border membrane vesicles isolated from rat kidney cortex. Uptake of SO(4) (2-) into brush-border membrane vesicles isolated from rat kindey cortex by a Ca(2+)-precipitation method was investigated by using a rapid-filtration technique. Uptake of SO(4) (2-) by the vesicles was osmotically sensitive and represented transport into an intra-vesicular space. Transport of SO(4) (2-) by brush-border membranes was stimulated in the presence of Na(+), compared with the presence of K(+) or other univalent cations. A typical ;overshoot' phenomenon was observed in the presence of an NaCl gradient (100mm-Na(+) outside/zero mm-Na(+) inside). Radioactive-SO(4) (2-) exchange was faster in the presence of Na(+) than in the presence of K(+). Addition of gramicidin-D, an ionophore for univalent cations, decreased the Na(+)-gradient-driven SO(4) (2-) uptake. SO(4) (2-) uptake was only saturable in the presence of Na(+). Counter-transport of Na(+)-dependent SO(4) (2-) transport was shown with MoO(4) (2-) and S(2)O(3) (2-), but not with PO(4) (2-). Changing the electrical potential difference across the vesicle membrane by establishing different diffusion potentials (anion replacement; K(+) gradient+/-valinomycin) was not able to alter Na(+)-dependent SO(4) (2-) uptake. The experiments indicate the presence of an electroneutral Na(+)/SO(4) (2-)-co-transport system in brush-border membrane vesicles isolated from rat kidney cortex.


Detection of a low-molecular-weight antigen on melanoma cells by a human antiserum in leukocyte-dependent antibody assays. Biochemical characterization of serologically detected human melanoma antigens was undertaken for the development of immunodiagnostic assays in melanoma. An antiserum from a human melanoma patient, which detected melanoma antigens expressed on a large proportion of different melanoma cells, was used in leucocyte-dependent cytotoxic antibody (LDA) 51Cr-release assays to monitor the purification of melanoma antigens in urea/acetate extracts of lactoperoxidase 125I-labelled melanoma cell membranes. The separation procedures included affinity chromatography on Concanavalin A, gel filtration on porous polyacrylamide beads and preparative isoelectric focusing. The fractions were also monitored by polyacrylamide electrophoresis in sodium dodecyl sulphate and by measurement of beta 2 microglobulin and carcinoembryonic antigen content. The antigens detected by this antiserum appeared to be acidic (pI 3.5) low-mol.-wt glycoproteins of approximately 15,000 daltons which were resistant to heating at 56 degrees C and digestion with neuraminidase, but susceptible to repeated freeze-thawing and trypsin digestion. They did not appear to be related to HLA antigens, beta 2 microglobulin or known foetal antigens. The nature of the antigens detected in these studies is as yet unknown, but they appear similar to those described in the sera and urine of melanoma patients in previous reports. Thes combined results and the frequent expression of these antigens on melanoma cells from different patients suggest that assays to detect this antigen may provide a valuable immunodiagnostic aid in the management of melanoma.


The relation between maternal plasma alpha-fetoprotein and birth weight in twin pregnancies. In the course of routine screening for neural tube defects, maternal plasma alpha-fetoprotein (AFP) was measured between 15 and 23 weeks of gestation in 64 twin pregnancies. Women with AFP levels more than twice the median for singleton pregnancies gave birth to infants with significantly decreased birth weights. Women with AFP less than the median also tended to produce twins with decreased birth weights. The distribution of gestations at delivery suggested that in twin pregnancies low AFP values gave an early warning of growth retardation while high values signal possible premature delivery.


Comparison of the effect of linear gramicidin analogues on bacterial sporulation, membrane permeability, and ribonucleic acid polymerase. Various analogues of linear gramicidin were tested for their biological activity in restoring the normal spore phenotype of gramicidin-negative mutants of Bacillus brevis and for their ability to increase cation conductivity of black lipid membranes and to inhibit bacterial RNA polymerase. Whereas many biologically active gramicidin analogues had no effect on membrane permeability, all biologically active peptides were able to inhibit ribonucleic acid (RNA) polymerase. These observations make it unlikely that membranes are the site of action of gramicidin during bacterial sporulation, but they are consistent with the notion that gramicidin functions to control RNA synthesis during the transition from vegetative growth to sporulation (Sarkar & Paulus, 1972). The relationship between peptide structure and the ability to restore normal sporulation and inhibit RNA polymerase showed that the eight amino-terminal residues have little influence on the function of gramicidin, whereas the highly nonpolar repeating sequence D-leucyl-L-tryptophan is essential for biological activity and may represent the site of interaction with RNA polymerase.


Adrenergic stimulation of membrane protein phosphorylation in human erythrocytes. Adrenergic modification of membrane protein phosphorylation was studied in intact human erythrocytes. Micromolar norepinephrine increased 32P incorporation into Band 2 by 70%, and into Band 3 by 40%. Phosphorylation levels observed with a series of specific agonists and antagonists suggest that an alpha-adrenergic receptor is involved in this effect. The mechanism of linkage between this receptor and protein phosphorylation does not appear to involve modulation of intracellular concentrations of ATP, cyclic AMP, or cyclic GMP.


Galactosyltransferase from commerical preparations of fetuin. A galactosyltransferase that transfers galactose from UDPgalactose to asialoagalacto fetuin or N-acetylglucosamine was partly purified from two commerical preparations of fetuin and its kinetic properties were characterized. Several other preparations of fetuin were also found to contain galactosyltransferase activity.


Role of cyclic GMP in submandibular gland secretion. Cholinergic stimulation of rat submandibular gland slices resulted in a rapid increase in the level of cyclic GMP. This increase was dependent upon the presence of Ca2+ and a phosphodiesterase inhibitor. Adrenergic agonists did not produce a significant elevation of cyclic GMP. The addition of Ca2+ to slices preincubated with the divalent ionophore A23187 caused a rapid rise in cyclic GMP levels which were unaffected by cholinergic stimulation. While these results could support a function for cyclic GMP in cholinergic-mediated K+-release from these glands, they do not support a role for this nucleotide in alpha-adrenergic agonist-induced K+-release or protease secretion.


[Immunochemical analysis of histone H1 and H5 from pigeon erythroid cells]. An antiserum with the antibody titer of 1 : 4096 was obtained by immunization of rabbits with the tRNA-histone H5 complex from pigeon erythrocytes. The specificity of the antiserum was studied quantitatively from the reaction of the complement binding to a homologous antigen (histone H5) and its modifications (I, II, III), differing in the degree of phosphorylation. It was shown that phosphorylation of histone H5 increases the ability of the antigen to bind to antibodies, which is especially well-pronounced at the antiserum dilutions as high as 20480. The comparison of the antigenic properties of histones H5 from pigeon and chicken erythrocytes revealed beside structural differences of the proteins the presence of common antigenic determinants. A similar observation was made when histones H5 and H1 from pigeon erythrocytes were compared. Histone H1 from chicken erythrocytes and histone H1 from calf thymus did not produce criss-cross reactions with antiserum H5.


[Anti-arrhythmic effect of tetrodotoxin in the late stage of experimental myocardial infarct]. The action of tetrodotoxin (TT), blocking the fast sodium current, on arrhythmias that occurred 24 hours after occlusion of the coronary artery was studied in 10 dogs. TT injected intravenously in doses of 0.5--3.0 micrograms/kg significantly decreased the number of ventricular extrasystoles, and completely restored sinum rhythm in 4 animals. The maximum antiarrhythmic effect was noted 3 to 5 minutes after TT adminstration. It is suggested that arrhythmias and the antiarrhythmic effect of the drugs in the late stage of myocardial infarction are connected with the fast inward sodium current.


[Effect of diiodobenzotefa on the functionally different subpopulations of T-lymphocytes]. Diiodobenzo-tepa (DIB) was given orally to CBA mice in a dose of 25 mg/kg for 3 successive days. The number of nucleus-containing cells decreased 3.9 fold in the thymus and 1.4 fold in the bone marrow. In experiments on transplantation of lymphoid cells to intact or lethally irradiated (CBA X C57BL/6J)F1 mice treated with DIB this substance did not influence the helper activity of T lymphocytes but inhibited the activity or B and T lymphocytes, inducing "graft-versus-host" and T cell-suppressor functions.


Subsets of malignant lymphomas in children related to the cell phenotype. We studied the lymphomatous cells of 39 children presenting with the classical features of malignant lymphoma. Twenty-two had T lymphoblasts. We could classify these patients into three subsets: The T lymphoblasts from children group 1 displayed antigen(s) shared by a thymocyte subpopulation, had terminal deoxynucleotidyl transferase (TDT), but no affinity for peanut agglutinin (PNA). The T lymphoblasts from children group 2 lacked the thymocyte antigen(s), had no TDT, but showed affinity for PNA. The T lymphoblasts from children group 3 displayed mature T-cell antigens, had no TDT, and no affinity for PNA. Children from the three groups were similar in terms of clinical presentation, age and sex distribution, and cell morphology; however patients from the three groups might have a different prognosis. Fourteen children had B lymphoblasts that, in half of the cases, had affinity for Helix pomatia agglutinin. Three patients had lymphoblasts lacking specific marker. Two of them had cells displaying an antigen found on common acute lymphoblastic leukemia cells and had TDT.


Plasma beta-thromboglobulin: differentiation between intravascular and extravascular platelet destruction. To elucidate the usefulness of beta-thromboglobulin (beta TG) in the differentiation of the mechanism of thrombocytopenia, plasma beta TG concentration was measured in one patient with amegakaryocytic thrombocytopenia, four patients with autoimmune thrombocytopenia (ATP), two patients with thrombotic thrombocytopenia (TTP), and one patient with thrombocytopenia secondary to disseminated intravascular coagulation (DIC). Plasma beta TG was not measurable in amegakaryocytic thrombocytopenia, was normal in ATP, and was increased in TTP and DIC. These data indicate that in thrombocytopenic patients, increased plasma beta TG concentration may result from intravascular platelet consumption with release of platelet constituents in contrast to extravascular platelet destruction by the macrophage-monocyte system.


A new fluorescence technique for staining of mononuclear phagocytes. A recently developed fluorochrome, 4',6-diamidino-2-phenylindole (DAPI), is used to stain mononuclear phagocytes of the mouse. After addition of heparin, these cells showed a bright yellow outer ring.


Methyl xanthines, adenosine 3',5'-cyclic monophosphate and the spinal transmission of nociceptive information. 1 In spinal cats anaesthetized with either alpha-chloralose or sodium pentobarbitone, a study was made of the effects of adenosine 3',5'-cyclic monophosphate (cyclic AMP), mono- and di-butyryl cyclic AMP and the methyl xanthines, theophylline and isobutyl methyl xanthine (IBMX), on the responses of neurones of laminae I, IV and V to noxious and innocuous skin stimuli. The compounds were administered from micropipettes positioned in the substantia gelatinosa. IBMX was also given intravenously.2 When administered in the substantia gelatinosa, neither cyclic AMP, its butyryl derivatives, nor the methyl xanthines had any effect on the excitation of neurones of spinal laminae IV and V by noxious heating of the skin or deflection of hairs. When the nociceptive responses of cells had been reduced by electrophoretic morphine, methyl xanthines and cyclic AMP failed to modify the effects of morphine on these deeper neurones. Electrophoretically administered naloxone reversed the effect of morphine.3 Intravenously administered IBMX (1 to 2 mg/kg) produced large transient increases in the firing rate of both C fibres and the excitation of dorsal horn neurones by noxious heating of the skin. These increases coincided with decreases in the mean systemic blood pressure, and probably resulted from increased temperatures being attained in the dermis by each noxious stimulus. When dorsal horn neurones were activated by electrical stimulation of the tibial nerve by a stimulus adequate to excite C fibres, intravenous IBMX produced a small or no increase in the number of spikes per stimulus.4 These results in the spinal cord do not support the hypothesis that the inhibition of synthesis of cyclic AMP is relevant to the analgesic action of morphine in mammals.


Non-immunological release of slow-reacting substance from guinea-pig lungs. 1 During incubation with calcium ionophore A23187, sensitized and unsensitized guinea-pig chopped lung released a slow-reacting substance (SRS) and histamine. 2 SRS possessed many characteristics of slow-reacting substance of anaphylaxis (SRS-A). It was inactivated by arylsulphatase, antagonized by FPL55712 (1 microgram/ml) and more stable in alkali than acid. Furthermore, it behaved like SRS-A  by stimulating archidonic acid metabolism in guinea-pig isolated perfused lungs. 3 Maximal ionophore generation of SRS in sensitized lung was greater than maximal anaphylactic generation of SRS-A but the release of histamine was not significantly different. Simultaneous challenge with antigen and ionophore produced more SRS-like or SRS-A-like activity than either stimulus alone. 4 These results have shown a non-immunological release from guinea-pig lung of SRS which was similar (or possibly identical) to SRS-A. It is suggested that in addition to mast cells, other cell types may be involved.


Axonal transport and metabolism of [3H]fucose- and [35S]-sulfate-labeled macromolecules in the rat visual system. The axonal transport of labeled macromolecules in retinal ganglion cells of rats was investigated from 1 to 20 days following intraocular injection of [3H]fucose and [35S]sulfate. Maximal incorporation of [3H]fucose into acid insoluble material in the retina was at 8 h, followed by a biphasic decline. Transported [3H]fucose (98% as glycoprotein) was in the optic nerve at 1 h, the optic tract and lateral geniculate body by 2 h, and the superior colliculus by 3 h after injection, indicating a rate of transport of approximately 200 mm/day. Radioactivity continued to accumulate in the superior colliculus for at least 8 h and began to decline rapidly by 24 h. Between 3 and 6 days levels rose again in both optic tract and superior colliculus before starting a gradual decline, indicating that a wave of rapidly transported material was delayed in leaving the retina. When proteins in the superior colliculus were fractionated by gel electrophoresis, the composition of the two fucosylated protein transport phases could be partially resolved. Radioactivity in individual gel peaks represented primarily in the first phase decayed with an average half-life of one day, althouth that in one prominent protein of molecular weight 280,000 turned over with a half-life of the order of 12 h. Radioactive peaks primarily in the second phase decayed with an average half-life of more than a week. Incorporation of [35S]sulfate into acid insoluble material in the retina was maximal at 1-2 h, after which there was a rapid loss of label. The appearance of [35S]sulfate in the optic tract, lateral geniculate body and superior colliculus preceded by a short time that of the [3H]fucose; indicating a shorter retinal processing time for this label. The total transported [35S]sulfate in the superior colliculus peaked by 4-8 h and had fallen by 65% at one day; no prominent second wave of transport was observed as was the case for [3H]fucose. Acid insoluble [35S]sulfate in the superior colliculus was equally divided between glycopeptides and glycosaminoglycans at all times examined, indicating that these macromolecules are transported at the same rate. [35S]Sulfate incorporated into various proteins fractionated by gel electrophoresis had heterogeneous turnover rates, the average being around 12 h. Radioactivity in one group of proteins, of molecular weight around 90,000, decayed with a half-life of only a few hours.


The uptake and radioautographical localization in the frog retina of [3H](+/-)-aminocyclohexane carboxylic acid, a selective inhibitor of neuronal GABA transport. The accumulation of [3H](+/-)-cis-3-aminocyclohexane carboxylic acid ([3H] ACHC) in frog retinae in vitro was highly localized in horizontal cells and their processes. [3H]GABA was also mainly accumulated within horizontal cells, but [3H]L-2,4-diaminobutyric acid ([3H]DABA) was taken up predominantly by the neuroglial Müller cells, whilst [3H]beta-alanine was localised largely within the photoreceptors. The uptake of [3H]ACHC (4.2 microM) was almost linear for 30 min and after 60 min a tissue/medium ratio of 5.25 was achieved. The uptake process was temperature sensitive highly dependent on sodium ions, and appeared to be mediated by a saturable transport process with an IC50 value of 0.83 mM. The accumulation of [3H]ACHC was inhibited by GABA and DABA (IC50 = 0.32 mM and 0.23 mM, respectively) whilst beta-alanine was a relatively weak inhibitor (IC50 = 9 mM) of ACHC uptake. In agreement with these results, the efflux of [3H]ACHC from the retina was increased by exposure to ACHC, GABA and DABA but not beta-alanine. In contrast, the efflux of [3H]DABA from the retina was not increased by GABA or ACHC, although DABA itself and potassium depolarization stimulated the release of [3H]DABA. These results strongly suggest that ACHC is accumulated in the frog retina by the same neuronal transport process as GABA. In contrast, the high affinity sites for DABA are localized mainly in glia, although inhibitor and release studies suggest that, at high concentration, DABA also interacts with the neuronal GABA (ACHC) transport process.


Prenatal and early postnatal beta-adrenergic receptor-mediated increase of cyclic AMP in slices of rat brain. The levels of cyclic AMP in slices of cerebral cortex and cerebellum from newborn rats were significantly, but transiently, increased by exposure to the beta-adrenergic agonist, isoproterenol. Isobutylmethyxanthine, an inhibitor of phosphodiesterase, enhanced this effect and permitted its detection in cerebral cortex obtained from the prenatal rat. These results are consistent with the possibilities that functional noradrenergic synapses are formed early in the ontogeny of the CNS, and that norepinephrine may exert cyclic AMP-mediated influences on brain development.


Organization of visual cortical projections to the claustrum in the cat. The projection patterns from different visual areas of the parieto-occipital cortex to the claustrum were studied autoradiographically in cats. When [3H]proline was injected into 17, 18, 19 or Clare-Bishop areas, the label was transported to an area restricted to the dorsal and caudal parts of the claustrum without any suggestion of retinotopic organization. Injection in each of these visual areas resulted in individual patterns of projection but with overlapping fields of termination, a pattern similar to corticocaudate projection. When injected into area 7, a region shown to have neurons involved in visuomotor mechanisms, the label was transported to the same area as that of the visual projection. These and other findings suggest that claustrum may be reciprocally and topographically connected with the cerebral cortex.


Projection of neostriatal spiny neurons to the substantia nigra. Application of a combined Golgi-staining and horseradish peroxidase transport procedure at both light and electron microscopic levels. One type of striatonigral neuron in the rat has been characterized. Golgi impregnation of striatal neurons that had been retrogradely labeled by horseradish peroxidase has shown that the medium-sized, densely spiny neurons project to the substantia nigra. Some of the synapses on three of these identified striatonigral neurons have been studied in the electron microscope following replacement of the Golgi deposit by means of the 'gold-toning' method. Synapsing axonal boutons were found on the following sites: soma and axon initial segment (symmetrical, with flattened or pleomorphic vesicles); primary and secondary dendritic shafts (symmetrical with pleomorphic vesicles); dendritic spines (asymmetrical, with spheroidal vesicles). These findings show that new information concerning neuronal connectivity can be obtained by combining three classical procedures in the same material: first, the Golgi method, that characterizes the type of neuron on the basis of its dendritic morphology; second, a retrograde tracing method, that identifies the projection area of the neuron; and, third, ultrastructural analysis of the nature of afferent terminals on the neuron.


Direct immunofluorescence tests with counterstains for detection of Chlamydia psittaci in infected avian tissues. Different methods of preparation and serological evaluation of rabbit globulins for use in fluorescent antibody conjugate and different methods of counterstaining with fluorescent antibody tests were evaluated for detection of Chlamydia psittaci in infected turkey tissues. The agar gel precipitin reaction was that chosen for testing and selecting antiserums to be used for fluorescein isothiocyanate conjugation. The fluorescent antibody staining was most pronounced with conjugate made from globulins precipitated with ammonium sulfate. A direct fluorescent antibody method with Evans blue counterstain correctly identified "coded" specimens of C. psittaci-infected and noninfected turkey air sacs. However, naphthalene black was superior to Evans blue as a counterstain when infected pericardial sacs were tested.


Surgical treatment of carcinoma of the prostate. Prostatic cancer kills more men than any other malignant condition except that arising from the lung. "Cancer Therapy: Prognostic Factors and Criteria of Response" predicted that there would be approximately 17 500 deaths due to this disease in the United States in 1978. Surgery is only applicable in stages A and B, when the tumour, as shown by various tests, such as measurement of the acid phosphatase value, technetium scanning and radioimmunoassay, is confined to the prostate. Ideally, the lymph glands along the iliac and obturator vessels should first be removed and quick-sectioned. If malignant cells are found in the lymph glands, the disease is considered surgically incurable and the procedure should be abandoned. If, however, the glands are disease free, a total prostatovesiculectomy should be carried out. The author also discusses the place of palliative surgery, such as transurethral resection, in the treatment of cancer of the prostate.


Some observations on serum gammaglobulin concentrations in suckled beef calves. Gammaglobulin concentrations were measured in serum samples collected from 23 single-suckled beef calves immediately after birth and from 346 calves at 48 +/- 12 hours postpartum. Considerable variation in these levels appeared to be associated with age and breed of dam. There were also indications that they might be influenced markedly by herd management. These post-suckling gammaglobulin concentrations were of no value in predicting incidence or severity of undifferentiated neonatal diarrhea in the beef herd as a whole, or in the individual neonate.


Kinetics of 57Co-bleomycin in mice after intravenous, subcutaneous and intratumoral injection. In tumor-free and tumor-bearing mice the body clearance and organ distribution of 57Co-BLM was measured in different time intervals after iv, sc, and it administration of the drug. No significant differences could be demonstrated in body clearances following different doses and routes of application of labeled BLM in tumor-free and tumor-bearing mice. The organ distribution studies showed higher concentrations following iv compared to sc or it application of 57Co-BLM; however, the activity in the ipsilateral injection sites were significantly increased after sc and it injection. In tumor-bearing mice the activity in the draining lymph nodes of the injection site were as high as that seen in draining lymph nodes following iv injection. However, on the contralateral side, the lymph node concentration was significantly reduced after it injection. These results indicate on the basis of organ distribution of 57Co-BLM a rational basis for it treatment of malignant tumors.


Focal nodular hyperplasia of the liver with alcoholic hyalin bodies and cytologic atypia. Focal nodular hyperplasia is an uncommon hepatic lesion in which the component hepatocytes are usually histologically normal. A case of focal nodular hyperplasia with cytologically atypical hepatocytes is presented. Light and electron microscopic evidence is given for the presence of alcoholic hyalin in these cells. A review of cytologic changes reported in focal nodular hyperplasia by others is included. To our knowledge, the presence of alcoholic hyalin in focal nodular hyperplasia has not previously been reported.


Primary leiomyosarcoma of the bone and its comparison with fibrosarcoma. Two cases of primary leiomyosarcoma of the bone are recorded, one in the distal fibula of a 66-year-old man, the other in the proximal tibia of a 61-year-old woman. The cytological, histological, and ultrastructural features of leiomyosarcoma of bone are described and compared with those of fibrosarcoma. These features are sufficiently characteristic to enable a confident diagnostic distinction between leiomyosarcoma and fibrosarcoma. Nevertheless, certain basic similarities exist between these two tumors, manifested at the ultrastructural level by the presence of myofilaments in fibrosarcoma; it would seem that the observed differences relate to the degree of development of the myofilamentous structures. It is postulated that primary leiomyosarcoma of the bone need not necessarily always arise from the media of blood vessels; it might also conceivably develop through advanced myogenic metaplasia of a sarcoma originating from fibroblastic tissue.


Inhibitory effects of 9-beta-D-arabinofuranosylguanine 5'-triphosphate and 9-beta-D-arabinofuranosyladenine 5'-triphosphate on DNA polymerases from murine cells and oncornavirus. The effects of the newly synthesized compound 9-beta-D-arabinofuranosylguanine 5'-triphosphate (ara-GTP) on the activity of DNA polymerases from mouse cells and oncornavirus were compared with those of 9-beta-D-arabinofuranosyladenine 5'-triphosphate. Ara-GTP did not replace deoxyguanosine 5'- triphosphate as substrate for these DNA polymerases but inhibited the activities of DNA polymerase alpha, beta, and gamma and viral DNA polymerase. DNA polymerase alpha was more sensitive than DNA polymerases beta and gamma and viral DNA polymerase to inhibition by ara-GTP. The inhibitions by ara-GTP and 9-beta-D-arabinofuranosyladenine 5'-triphosphate were due to competition or partial competition 5'-triphosphate were due to competition or partial competition with deoxynucleoside triphosphate with the same base. The inhibition constant (Ki) and the mode of inhibition of nucleotide incorporation varied depending on the combination of inhibitor, substrate(s), and enzyme species.


Detection of both T-cell and Ia-like antigens on cells from patients with acute myelomonocytic leukemia and chronic myelogenous leukemia in blast crisis. Appropriately absorbed antisera to the lymphoblastoid cell lines HSB and SB detect a human T-lymphocyte-associated antigen (TLAA) and the human Ia-like antigens, respectively. Cells from some patients with acute myelomonocytic leukemia (AMML) and chronic myelogenous leukemia in blast crisis expressed both TLAA and Ia antigens when tested in a complement-dependent microcytotoxicity assay (greater than 90% lysis with both antisera). When patients were in remission, expression of TLAA and Ia antigens returned to normal values. Quantitative absorption of anti-TLAA serum with increasing numbers of AMML cells showed that these cells could remove reactivity of the serum for both HSB and human thymocytes. Similarly, absorption of anti-Ia serum with AMML cells removed all serological reactivity when this serum was tested on chronic lymphocytic leukemia cells or normal B-cells. These serological findings were confirmed by immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis studies using radiolabeled antigens. Cells from an AMML patient were labeled with 125I using lactoperoxidase; both the TLAA and Ia antigens were precipitated from the resulting solubilized membrane preparation. Leukemic cells from one AMML patient and one patient with chronic myelogenous leukemia in blast crisis were studied for Ia and TLAA antigens with a double fluorescence technique. Over 80% of the cells showed dual fluorescence.


Mutagenicity and cytotoxicity of nineteen heterocyclic mustards (ICR compounds) in cultured mammalian cells. The mutagenicity and cytotoxicity of 19 ICR compounds, including 6 reported previously, have been determined in the Chinese hamster ovary/hypoxanthine-guanine phosphoribosyltransferase system. As with other physical and chemical agents, ICR 170 and 191 exhibit a phenotypic expression time of 7 to 9 days, independent of concentrations tested. Thirteen of these compounds are mutagenic. At equimolar concentrations, the compounds with the tertiary amine-type side chain (ICR 217, 340, 355, 368, 170, and 292) are more mutagenic than the compounds with the secondary amine-type side chain (ICR 449, 371, 191, and 372). All secondary amine types show a "plateau" in their concentration-dependent mutagenesis curves at 3 to 4 microM. Shortening of the side chain by one carbon (ICR 171) results in a reduced mutagenicity. Substitution of a sulfur atom for a nitrogen in the side chain (ICR 342) increases both mutagenicity and cytotoxicity. The presence of two 2-chloroethyl groups on the side chain (ICR 220) also results in greatly increased cytotoxicity and mutagenicity. When the 2-chloroethyl group of ICR 340, 372, 292, 191, or 170 is replaced by a 2-hydroxyethyl group (ICR 340-OH, 372-OH, 292-OH, 191-OH, or 170-OH), a mutagenically inactive compound results which remains toxic. Replacement of the amine linkage with an ether linkage (ICR 283) also yields a mutagenically inactive compound.


Role of single-breath carbon monoxide-diffusing capacity in monitoring the pulmonary effects of bleomycin in germ cell tumor patients. Serial pulmonary function tests including single-breath carbon monoxide-diffusing capacity (DLCO), forced vital capacity (FVC), and forced expiratory volume in 1 sec were performed in a relatively homogeneous group of male patients with germ cell tumors treated with vinblastine, bleomycin, and cis-diamminedichloroplatinum. Of the pulmonary function tests used, the DLCO was shown to be the most sensitive indicator of subclinical bleomycin pulmonary effects. Decreases in DLCO were both total dose and schedule dependent. Patients receiving their total dose of bleomycin at a rate of 25 +/- 2 (S.D.) units/week developed a linear decrease in DLCO with increasing total doses of bleomycin. Changes in FVC did not correlate with bleomycin total dose. Although both the mean DLCO and FVC decreased after completion of bleomycin therapy, the mean FVC returned to base-line levels rapidly, whereas the decrease in mean DLCO was persistent for several months. When routine volumetric tests (FVC and forced expiratory volume in 1 sec) and DLCO are used in a systematic manner, DLCO is the most sensitive indicator of the subclinical pulmonary effects of bleomycin in germ cell tumor patients treated with vinblastine, bleomycin, and cis-diamminedichloroplatinum.


Control of permeation of bleomycin A2 by polyene antibiotics in cultured Chinese hamster cells. Control of permeation of bleomycin A2, a well-known antitumor antibiotic, in combination with various polyene macrolide antibiotics was analyzed in cultured Chinese hamster cells in vitro. Three polyene antibiotics, filipin, pentamycin, and pimaricin, were found to enhance the action of bleomycin A2 remarkably, while amphotericin B or nystatin could not. Although DNA synthesis and colony-forming activity of polyene-sensitive Chinese hamster V79 cells were synergistically inhibited by the combination of filipin and bleomycin A2, in a polyene-resistant subline (AMBR-1) derived from V79, they were only slightly affected in the presence of both drugs. The cellular uptake of [14C]bleomycin A2 by V79 was enhanced 2- to 4-fold in the presence of increasing doses of filipin or pentamycin, but not in the presence of amphotericin B. The treatment of V79 cells with filipin for 20 to 30 min was enought to block DNA synthesis almost completely when combined with 20 microgram belomycin A2 per ml. The pretreatment of the hamster cells with 6 microgram filipin per ml for 60 min continued to enhance the inhibitory action by bleomycin A2 of DNA synthesis up to 5 hr after the removal of filipin from the cultured medium.


Staging procedures and the role of surgery in testicular cancer. The conventional staging system for testicular cancer is inadequate in discriminating between varying degrees of local, nodal, and pulmonary disease. This staging system which uses conventional tests including lymphangiogram, iv pyelogram, and inferior venacavogram has been demonstrated to have a 35%--53% error rate in distinguishing between stages I and II cancer based upon retroperitoneal node dissection. A surgicopathologic staging system has been proposed which improves upon the conventional staging system for testicular cancer. This clinical and surgicopathologic staging system has been proposed based on determination of serum alphafetoprotein and human chorionic gonadotropin before and/or after orchiectomy and lymphadenectomy with a low staging error. The conventional surgical management of testicular cancer and the current status of the National Cancer Institute prospective randomized clinical trial assessing the role of intensive chemotherapy with or without cytoreductive surgery in poor-risk bulky stage III testicular cancer are discussed.


Applications of biologic tumor markers in testicular cancer. Quantitative measurement of serum alphafetoprotein (AFP) and human chorionic gonadotropin (HCG) by double antibody radioimmunoassays originally developed at the National Cancer Institute, Bethesda, Md) has been performed in 130 patients with seminoma and in 300 patients with nonseminomatous testicular cancer. Ten of 130 (7.7%) patients with seminoma, 99 of 152 (65%) patients with embryonal carcinoma with or without teratoma, and five of five (100%) patients with pure choriocarcinoma had an elevated level of HCG. None of the patients with seminoma or choriocarcinoma had an elevated level of serum AFP although elevated levels were present in 67% of the patients with embryonal carcinoma with or without teratoma. Either HCG or AFP was elevated in about 90% of the patients with clinically demonstrable embryonal carcinoma with or without teratoma and choriocarcinoma. These two markers have been helpful in staging, detecting recurrence, prognosis, and localization (by the alpha subunit of HCG) of a tumor. Utilizing immunocytochemical techniques, HCG has been localized in syncytiotrophoblastic component of choriocarcinoma. AFP has been localized in yolk sac tumor and embryonal carcinoma cells.


Vinblastine, bleomycin, and cis-dichlorodiammineplatinum(II) in disseminated testicular cancer: preliminary report of a Southwest Oncology Group Study. Combination induction chemotherapy consisting of vinblastine, bleomycin, and cis-dichlorodiammineplatinum(II) was administered to 126 patients with advanced germ cell tumors of the testis. Following this 4-month induction therapy, maintenance treatment consisting of chlorambucil and actinomycin D alternating monthly with vinblastine was given for an additional 20-month period. Sixty-four patients (51%) achieved a complete remission, while 33 patients (26%) achieved a partial response. Responses were observed in all histologic categories. Seven patients (11%), who were in complete remission, have relapsed at a median time of 5 months. Median duration of remission is greater than 12 months for complete responders and 4 months for partial responders. Significant leukopenia was noted in 57% of evaluable courses and significant thrombocytopenia was noted in 18%. Less than 20% of patients were noted to have nephrotoxicity. Other toxic effects observed were gastrointestinal (42%), dermatologic (15%), pyrexia (14%), neuromuscular (9%), and pulmonary (less than 5%). Several pretreatment parameters are currently being evaluated to determine the impact on complete remission rate and survival.


Treatment of metastatic nonseminomatous testicular cancer: a preliminary report of induction chemotherapy followed by maintenance chemotherapy or radiotherapy. Patients with metastatic nonseminomatous testicular cancer received an induction regimen consisting of bleomycin in 24-hour infusions and bolus iv doses of vinblastine followed by an Adriamycin and cis-dichlorodiammineplatinum(II) combination. Patients achieving complete remission after one or two cycles of this induction chemotherapy were then randomized to receive either radiotherapy (RT) to the previously involved tumor areas or maintenance chemotherapy (MCT) with CCNU, methotrexate, and vinblastine for 2 years. Among 62 evaluable patients, induction chemotherapy achieved 15 (24%) partial remissions and 35 (56%) complete remissions. Two patients with partial remission and single pulmonary metastases were rendered disease-free by surgical resection of residual tumor. Twenty patients received MCT and 15 received RT. To date, median survival is 10,8+ months in the MCT group with five relapses and 12.5 months in the RT group with two relapses. Toxicity in the induction phase was moderately severe with two drug-related deaths.


Chemotherapy for advanced germinal cell neoplasms: preliminary report of an Eastern Cooperative Oncology Group Study. A series of 45 evaluable patients with stage III germ cell tumors have been treated between July 1977 and May 1978, with two cycles of an intensive five-drug regimen including cyclophosphamide, actinomycin D, vinblastine, bleomycin, and cis-dichlorodiammineplatinum(II) (cis-platinum). The patients were randomly assigned to receive the cis-platinum as a high dose 1-hour infusion with mannitol diuresis or as a low-dose 8-hour infusion without diuresis. The overall response rates were 36% complete response (CR) and 47% partial response (PR) after two cycles of induction therapy. Patients who achieved a CR or PR after two cycles were randomized to continue treatment with either a three-drug program of vinblastine, actinomycin D, and cis-platinum or a cyclic crossover regimen of three regimens (each containing three drugs) given in sequence at 3-week intervals with the entire sequence repeated at 9 weeks. During continuing treatment a minimum of five patients improved from PR to CR, so that the minimum CR rate will be 47%. A major determinant of response was extent of disease, with limited-disease patients having an 83% CR rate and advanced-disease patients having 22% CR rate. At present, there are no statistically significant differences between response rates for the two induction regimens. Patients have not been followed long enough on the continuing treatment arms to allow evaluation of improvement from PR to CR, duration of response, or duration of survival. Hematologic and renal toxiciity were not significantly different on the two induction treatment arms.


Sequential combination chemotherapy and surgery for disseminated testicular cancer: cis-dichlorodiammineplatinum(II), vinblastine, and bleomycin remission-induction therapy followed by cyclophosphamide and adriamycin. A multimodality treatment program for disseminated testicular cancer using vinblastine/bleomycin/cis-dichlorodiammineplatinum(II) remission-induction therapy followed by surgery and cyclophosphamide/Adriamycin maintenance therapy has been used in 25 patients. The overall complete and partial response rate was 100%. Six of six patients (100%) with stage II (advanced abdominal) disease have remained in complete remission for a median followup period of 8 months. Seventeen of 19 patients (89%) with stage III metastatic disease have been in complete remission for a median duration of 20 months. No relapses have occurred in any patients who achieved a complete remission. Two of 19 patients (11%) with stage III disease achieved a partial response; one died at 4 months and the other died at 14 months. The toxicity was severe, especially during remission induction. However, there were no drug- or surgically related deaths.


Connectivity of the auditory forebrain nuclei in the guinea fowl (Numida meleagris). Injection of tritiated leucine and proline into the nucleus ovoidalis of the Guinea Fowl (Numida meleagris) produces terminal labeling in the palaeostriatum and in three adjacent zones (field L1-L3) of the auditory neostriatum (AN). L2, situated between L1 and L3, receives the main input and corresponds to the former field L of Rose. These neuroanatomically defined zones of the auditory neostriatum are also characterized by differing properties of their neurons. Injection of radioactive material into the auditory neostriatum produces labeling of (i) a palaeostriatal, (ii) a ventral hyperstriatal, and (iii) an additional neostriatal area (Nd). Injection into the hyperstriatum ventrale reveals connections (i) to field L2, (ii) to the palaeostriatum, (iii) to Nd, and (iv) to the archistriatum. After injection into the palaeostriatum, labeling can be observed (i) in the neostriatum dorsale, (ii) in the hyperstriatum ventrale, (iii) in the archistriatum, (iv) in the diencephalic nuclei, nucleus ansae lenticularis and nucleus spiriformis lateralis, and (v) in the mesencephalic nuclei, nucleus tegmenti pedunculo-pontinus and nucleus intercollicularis. These results show that a widespread connectivity exists among primary and presumably higher order auditory areas in the forebrain of birds. Connections also exist between these auditory areas and presumed vocal-motor areas (neostriatum dorsale, archistriatum, nucleus intercollicularis).