Carcinoma of the esophagus: results of treatment. The treatment results of the Rotterdam working group on esophageal cancer during the period January 1970-January 1978 were assessed. A total number of 328 patients were treated: 230 males and 98 females. Of the 133 patients eligible for a combined treatment modality i.e. preoperative radiotherapy and surgery, 52 showed irresectable or metastatic disease during operation. The five year actuarial survival rate of the 81 patients, in whom curative surgical resection of the tumor was performed, amounted to 21%. Females fared better than males, the five year survivals being 42% and 12% respectively. This female preponderance in survival is partly explained by the considerable postoperative mortality of the male patients: 28% vs 7.4% in females. Patients who received only radiation therapy, whether curative or palliative, had a very bad prognosis. It is concluded that preoperative irradiation followed by surgical removal of the tumor should be performed in all operable-curable patients.


Esophagogastrectomy for carcinoma: current hospital mortality and morbidity rates. Between January 1, 1970, and March 1, 1979, 153 patients with carcinoma of the esophagus or cardia were seen at the Lahey Clinic; 124 (81%) underwent surgical exploration and 102 (82.3%) were found amendable to resection. This report concerns the 82 patients operated on by the senior author, 72 of whom (87.8%) had surgical resection. A variety of resective techniques were used but currently esophagogastrectomy and esophagogastrostomy is preferred, a left thoracotomy being used for low lying lesions; upper thoracic and cervical lesions are approached through a combined abdominal and right thoracic approach or esophagectomy with cervical esophagogastrostomy and without thoracotomy is used. Two deaths occurred within 30 days of operation, a hospital mortality rate of 2.8%. Significant complications developed in 11 patients (15.3%). The average survival was 20.8 months, and satisfactory long-term relief of dysphagia was achieved in 91.2% of patients. An aggressive surgical approach to the management of patients with carcinoma of the esophagus or cardia is justified, for esophagogastrectomy and esophagogastrostomy is applicable to the majority of patients; can now be performed at low risk with a reasonable period of hospitalization; and provides satisfactory long-term palliation.


Sex-hormone-binding globulin in the plasma of normal men and patients with benign prostatic hypertrophy. Sex-hormone-binding globulin (SHBG) was measured in the plasma of normal men aged 30--50 yr, normal men 50--70 yr of age, and men 50--70 yr of age suffering from benign prostatic hypertrophy (BPH). The level of binding proteins in normal "young" men decreased approximately 45% in normal "older" men. A 100% rise was observed in BPH patients as compared to normal older men. This increase seemed to be related to a rise in SHBG and also to the appearance of binding species of smaller molecular weight than that of SHBG.


Tuberculosis. Clinical aspects and diagnosis. The presentation of tuberculosis is variable depending on the severity of the infection, the age of the patient, whether the infection is primary or secondary, and whether the manifestations are due to inhalation of organisms or hematogenous dissemination. A definitive diagnosis is made by culture of the organism; spontaneously expectorated sputum is the most suitable specimen for diagnosing pulmonary tuberculosis. Diagnosis of extrapulmonary tuberculosis frequently requires tissue biopsy. The classic staining method for demonstrating tubercle bacilli is the Ziehl-Neelsen technique. Newer methods based on fluorescent dyes and phase-contract microscopy make rapid screening feasible, but false-positive identification is more frequent. Culture of tubercle bacilli is most successful when two media are used. The differential diagnosis of pulmonary tuberculosis includes bacterial pneumonia, especially anaerobic infection, and fungal infections including histoplasmosis, coccidioidomycosis, and blastomycosis. Lung carcinoma can mimic tuberculosis and the two diseases can coexist. Surgery is frequently necessary for a definitive diagnosis, expecially when the disease is seen as a noncalcified nodule.


Regulation of the kidney xanthine dehydrogenase by glutaminase. The steady-state concentrations of glutamine, glutamate and ammonia in the kidney cells might regulate the rate of renal xanthine dehydrogenase activity. Both glutamate and glutamine were found to be effective inhibitors of the renal xanthine dehydrogenase activity in vivo. The inhibition by glutamate depends essentially on the glutaminase inhibition.


Pattern of the ventilatory response to transient hypoxia in man: differences from transient hypercapnic test. The pattern of change in ventilatory variables after inhalation of pure N2 for two breaths was studied in normal children and adults. In six subjects the trends of change were compared to the ventilatory response to transient hypercapnia. We observed differences in the patterns of increasing ventilation with an initial abrupt increase of tidal volume for transient hypoxia and a progressive change for hypercapnia. In both cases respiratory frequency was progressively but unsystematically enhanced. A highly significant positive correlation was demonstrated between individual sensitivities to CO2 and O2, with a greater response to hypercapnia (5.6 time) than to hypoxia. Finally, a very short-latency decrease in expiratory duration occurred in the first breath after inhalation of hypercapnic mixture, supporting the recent data of Cunningham et al. (1977).


alpha-Methyl-D-glucoside uptake in renal cortical slices of normal and alloxan diabetic rabbits. The uptake of alpha-methyl-D-glucoside by slices of renal cortex was compared in normal and alloxan diabetic rabbits. Alloxanized rabbit tissue showed significantly higher levels of sugar accumulation than normal tissue. Diamide, which is known to oxidize intracellular glutathione (GSH), inhibited the uptake of alpha-methyl-D-glucoside by renal cortical slices. GSH stimulated sugar uptake and was also capable of reversing the inhibition of sugar accumulation caused by diamide. Mercaptoethanol and dithiothreitol, which are commonly used thiol compounds, were not as effective as GSH in stimulating sugar uptake. The level of GSH found in normal and alloxan diabetics rabbit kidneys shows a slightly decreased cortical GSH content in alloxanized animals. One can conclude that GSH participates in sugar uptake in kidney slices from both diabetic and normal rabbits.


[Effect of S-adenosyl-L-homocysteine of dopamine catabolism]. 1.--The administration of SAH to rats, at physiologically active dose on the sleep, does not change the urinary level of MD and NM. On the other hand, the excretion of DA and NA decreases. 2.--In the brain, SAH does not modify neither the concentration of NA and NM in hypothalamus and thalamus, nor the concentration of DA and MD in corpus striatum. 3.--After intracisternally injection of [14C]DA or [3H]NA, SAH increases the level of [14C]MD and [3H]NM. 4.--Contrary to the studies in vitro, where SAH is an inhibitor of COMT, on the rat it does not seem prevent the methylation of DA and NA.


Quantitative measurement of gluconeogenesis from isobutyrate in sheep. Experiments with continuous infusion of [14C] isobutyrate and single injection of [3H] glucose were performed in two sheep under fed and fasted conditions in order to investigate the contribution of isobutyrate to glucose synthesis. The pool size, total entry and irreversible loss of glucose in the fed sheep were 2.8 mmol/kg0.75, 1.70 and 1.43 mmol/h per kg0.75. After 72-h fasting these parameters decreased about 40% but recycling of glucose carbon increased from 16 to 38% of the total entry rate. Isobutyrate infused intravenously at a rate of 3.5 mmol/h contributed to a minimum of 3-5% of glucose entry indicating that at least 40-60% of the infused isobutyrate was used for net glucose synthesis. The efficiency of the glucogenic and energetic use of isobutyrate as compared to propionate is discussed.


Biological parameters of the blood in the genetically obese Zucker rat. A study of the various biological parameters of the blood in the genetically obese Zucker rat, the nonobese Zucker rat, and the Wistar rat has revealed great similarity between the two latter types of animals. On the other hand, in genetically obese Zucker rats as compared with the nonobese ones, (1) the blood mass per unit of weight was lower; (2) the level of nitrogenous degradation compounds was the same; (3) the lipase activity was lower; (4) the levels of substances for which liver plays a crucial role--all lipid and protein fractions, glucose, and the enzyme GPT--were higher; (5) the levels of Ca, Zn, Fe, Cu and Pi were high; (6) the blood and bone-marrow cells were unremarkable.


Ouabain-sensitive and -insensitive ATPase activities in the kidney and the liver of spontaneously hypertensive rats. Ouabain-sensitive and-insensitive ATPase activities were measured in the kidneys and the kidneys and the liver of male and female rats of the Lyon hypertensive (LHS) and Lyon normotensive strain (LNS). At the age of 48 +/- 2 weeks, hypertensive rats exhibited a significantly higher blood pressure and body weight than age-matched normotensive rats. Ouabian-insensitive ATPase activity was decreased in the kidney, but not in the liver of hypertensive rats compared to normotensive rats. Ouabian-sensitive ATPase activity was non significantly decreased in the kidney of hypertensive rats.


Effects of nickel ion on the conduction of action potentials in non-myelinated nerve fibre of crayfish. Effects of NiCL2 and PCMB (p-chloromercuribenzoate) on the action potential were examined by the method of extra- and intracellular electrodes, using a single nerve fibre of crayfish. The results obtained were as follows : The conduction of the action potential was blocked by treating the nerve fibre with Ni ion or PCMB. The blockade was easily recovered by replacement with cysteine. The process of the blockade and recovery, which could be repeated several times, was fairly characteristic such that the more repetition led the sooner blockade and the harder recovery. No conduction block was observed by treatment with Ni-cysteine mixed solution nor with PCMB-cysteine solution. The critical concentration for blocking was 1.1 x 10(-4)M for NiCL2 and 5.6 x 10(-6) M for PCMB. The action potential was disappeared without any change in the resting potential by treatment with the chemicals, which gave significant effects on the rising and falling phases of the action potential before the blockade.


Prostaglandins do not affect ion concentrations in human erythrocytes. In an attempt to further our understanding of the mode of action of prostaglandins (PGs), experiments were performed in which sodium and potassium ion concentrations were measured in human erythrocytes exposed to various concentrations of PGs. No consistent effect was observed and it is concluded that such changes are not likely to underlie excitability alterations induced by PGs.


Concentrating capacity of the kidney and nitrogen partition in the urine in Perodicticus potto (Prosimii, Lorisidae, Lorisinae). The concentrating ability of the kidney in Perodicticus potto was investigated. As the relation between this capacity and the thickness of the medulla has been stressed by Schmidt-Nielsen & O'Dell, the size of the various layers in the kidney was measured. Based on our observation that the potto may excrete uric acid in fairly high concentrations, the partition of nitrogen in the urine was also explored.


Relatedness among coagulase-negative staphylococci: deoxyribonucleic acid reassociation and comparative immunological studies. DNA-DNA-homology values were determined under restrictive to relaxed reassociation conditions with type strains and some additional strains of coagulase-negative staphylococci belonging to ten different species. The immunological relationship of the catalases present in the type strains of these species was also determined by applying double immunodiffusion and microcomplement fixation. The results of these studies support the previous proposal to subdivide the coagulase-negative staphylococci into at least ten separate species. However, it is evident that some of the species are more closely realted than others and can form species groups. According to the results presented in this study, the coagulase-negative staphylococci can be combined into five species groups: The Staphylococcus saprophyticus group is composed of S. saprophyticus, S. xylosus and S. cohnii. The S. epidermidis group comprises S. epidermis, S. capitis and S. warneri. The S. hominis group which exhibits a significant relationship to S. epidermidis includes S. hominis and S. haemolyticus. The species group S. sciuri consists of S. sciuri ssp. sciuri and S. sciuri ssp. lentus and the species group S. simulans is presently represented by the corresponding single species.


Carcinoembryonic (CEA)--like antigen in adenomas and cancers of the breast in human patients. By using specific anti-CEA serum and indirect immunofluorescence, a substance was detected in breast cancers which reacted similarly to CEA demonstrable by the same method in cancers of gastrointestinal tract. However, the double diffusion test in agar gel and additional absorption analysis showed that it was a glycoprotein different from CEA, although possessing common or similar antigenic determinants with it. This antigen also differs from another antigen called "nonspecific cross-reacting antigen NCA" and from alpha 1-acid glycoprotein of human serum.


Immunoglobulins of colostrum. V. Comparative studies of ovine and bovine serum and colostral IgM. Comparative studies of physiocochemical properties of ovine and bovine serum and colostral IgM are described. Serologic properties, molecular weight, amino acid composition, carbohydrate content, peptide maps, heterogeneity, and ORD and CD spectra were compared. Colostral IgM have additional antigenic determinants, higher carbohydrate content, lower heterogeneity, and higher content of beta-structure. Different amino acid composition and different peptide maps of serum and colostral IgM were shown. The results obtained suggest that ovine and bovine colostral IgM immunoglobulins may be a mixed population of molecules locally synthesized and transferred from the serum.


Antigenic structure and biological characteristics of influenza strains isolated in the USSR during the 1974/75 epidemic. Antigenic analysis of strains isolated in the USSR during the epidemic in 1974/1975 showed differences in the hemagglutination inhibition test and in neuramidase activity with antisera against three reference strains. Strains isolated in a later period of the epidemic were classified into subgroup A/Port Chalmers/1/73. Nearly all strains were effective inducers of interferon and were susceptible to this inhibitor of virus replication.


Antigenic similarity of morphologic type CIII1 bacteriophages. The aim of the study was to demonstrate antigenic differentiation among bacteriophages belonging to the same morphologic type--CIII1 according to Krzywy and Slopek or A2 according to Ackermann. Twenty-six bacteriophages which multiplied on various strains of bacteria of the genera Escherichia, Shigella and Klebsiella, were studied. Serologic tests were done by the quantitative complement fixation test. Immune sera against 5 phi E. coli. D8 Sh. flexneri, G35 Sh. sonnei and Kl7 Klebsiella bacteriophages were obtained from rabbits. It was shown that bacteriophages of CIII1 morphologic type, with morphologically identical virion had different antigenic specificity. On the basis of results obtained the phages could be divided into eight serologic groups. The bacteriophages for which antisera were obtained belonged to four different serologic groups. All bacteriophages had some common antigens with 5 phi E. coli phage. The similarity points to a phylogenetic relationship among phages of CIII1 morphologic type.


HLA gene and haplotype frequencies in the population of southern Poland. Results of a population study on frequencies of phenotypes and genotypes of the HLA system (locus A and B) in 240 adults, unrelated and of both sexes, living in southern Poland are reported. Frequencies of haplotypes were calculated according to Mattiuz et al. Distribution of all HLA specificities conformed with the Hardy-Weinberg law and was similar to the distribution in other white populations, except HLA-B13 which had a phenotype frequency of 14.17% and gene frequency 7%. Haplotype frequencies and positive and negative association were analogous with those in other white populations.


Isolation and cell culture propagation of rotaviruses from turkeys and chickens. Rotaviruses were detected by electron microscopy in the faeces of turkey poults and broiler chickens with diarrhoea. Apparently symptomless infection was also observed in broilers. The avian rotaviruses could not be isolated in cell cultures by conventional techniques, but were adapted to serial growth in chick cell cultures following trypsin treatment of the virus and the cells. Immunofluorescence studies showed that the avian and mammalian rotaviruses are antigenically related. Antibodies to rotavirus were widespread in sera collected from turkeys, chickens and ducks.


C-type RNA virus from hamster cells transformed in vitro by 1-B-D-arabinofuranosylcytosine. Evidence of type-C RNA viral activity in fetal hamster cells transformed in vitro by 1-B-D-arabinofuranosylcytosine (ara-C) after at least one in vivo passage is described. The virus possesses properties typical of other type-C RNA viruses, such as: a) morphology as determined with the electron microscope, b) presence of 70S RNA, c) enhanced expression following treatment with halogenated pyrimidines, d) group specific antigens of hamster type, and e) a buoyant density of 1.15 g per cm3. However, the virus particles are deficient in RNA-dependent DNA polymerase activity under conditions that easily detect Rauscher Leukemia virus and will infect neither hamster, rat, mouse, human nor rabbit cells. The possible role of this virus in chemical carcinogenesis of cultured hamster fetal cells is discussed.


[Cryptococcosis with a lesion of the meninges and adrenals (on the choice of fungal stain in histological sections]. A description of vitally unrecognized cryptococcal meningitis and tumour-like cryptococcosis of the adrenals is presented. Comparison of the results of various stainings for degeneratively changes and phagocytized fungi suggests that cryptococci in histological sections should be stained by Zihl-Nilsen's method (methylene blue and basic fuchsin).


[Excretion of urinary hydroxyproline in urinary tract diseases]. Urinary hydroxyproline has been measured in 150 patients, 70 of them with urinary diseases. High values were found in chronic hyperazotaemia and in carcinoma with bone metastasis. In the latter, the prognostic value was of particular interest where the lesions are not yet identifiable radiologically.


A modified filtering operation for neovascular glaucoma. Thirteen patients with useful vision but suffering from uncontrollable neovascular glaucoma due to total angle closure were treated with modified filtering operations. Ten (77%) of the cases were controlled more than one year postoperatively. Most patients whose pressures were controlled maintained their preoperative visual acuities. These visual results appear to be better than those reported with cyclocryotherapy, where vision is usually lost despite satisfactory pressure control.


Iris neovascular tufts. Relationship to rubeosis, insulin, and hypotony. Iris neovascular tufts are abnormal proliferations of twisted microcapillary loops that project from the iris pupillary border. They appear to be a morphologic variant of rubeosis iridis. This study searched for iris neovascular tufts by biomicroscopy and iris fluorescein angiography. They were found in two of 16 (12.5%) myotonic dystrophy patients, in two of 30 (6.7%) maturity-onset diabetics, and in zero of 14 (0%) juvenile-onset diabetics. Myotonic and adult-onset diabetics have in common pancreatic beta-cell dysfunction, abnormally high serum insulin levels, and possibly defective hormone-receptor interactions. Whether these factors contribute to the altered blood-iris barrier and iris neovascular tufts in these groups is uncertain, but a correlation is suggested because the myotonic with the highest insulin level also had the most prominent tufts. Hyperinsulinemia and ocular hypotony are associated with neovascular tufts.


Genetics of formamidase-5 (brain formamidase) in the mouse: localization of the structural gene on chromosome 14. A single formamidase, which is different from the formamidases found in other tissues occurs in the brains of mice. This enzyme is here called formamidase-5 and the gene symbol is designated For-5. Two alleles are recognized on the basis of their differential heat sensitivity:For-5b is relatively heat stable and is present in strain C57BL/6J, while For-5d is relatively heat sensitive and is present in strain DBA/2J. The heat sensitivity of formamidase-5 in 44 other inbred strains and substrains was tested and found to resemble that of C57BL/6J or DBA/2J. Thirty-six recombinant inbred strains derived from progenitors that differed at For-5 were studies to test for single-gene inheritance and linkage with other loci. Complete concordance was found with the esterase-10 locus (Es-10), indicating close linkage. The 99% upper confidence limit of the distance between For-5 and Es-10 is 3.7 centimorgans (cM). Es-10 is located on chromosome 14 about 19 cM from the centromere. An independent demonstration of linkage of For-5 with Es-10 and another chromosome 14 marker, hairless (hr), is provided by the finding that the HRS/J strain, which has been sibmated for 60 generations with forced heterozygosity at the hr locus, is cosegregating at For-5 and Es-10. A survey of 32 inbred strains and substrains revealed that the For-5d allele is associated with the Es-10b allele, and that the For-5b allele is associated with Es-10a and Es-10c. Formamidase-5 segregates as expected in the F2 generation of crosses between strains bearing For-5b and For-5d alleles. It is possible that this unique formamidase of the brain is involved in the metabolism of a neurotransmitter substance.


Further studies on the structural requirements for polypeptide-mediated histamine release from rat mast cells. Structure-activity studies have been performed on a series of naturally occurring and 'tailor-made' polypeptides, by measurement of ability to induce selective histamine release from normal rat peritoneal mast cells in vitro. Compounds investigated include corticotropin and melittin derivatives, mast-cell-degranulating peptide from bee venom, polymyxin B, bradykinin and various synthetic poly(amino acids) and short-chain peptides. It was confirmed that a cluster of four basic residues (lysine or arginine) was optimal for histamine release by corticotropin and melittin polypeptides, provided that the C-terminal carboxyl group was substituted (by, for instance, amidation). In contrast, the presence of a free C-terminal carboxyl group or nearby dicarboxylic acid residues led to a considerable diminution in histamine-releasing activity. Likewise, polypeptides comprised essentially of acidic amino acids were inactive. On the basis of these observations it has been possible to predict that synthetic peptides comprising a particular sequence within the Fc region of human immunoglobulin E, the immunoglobulin class particularly involved in mediation of allergic reactions of the immediate type, would possess potent histamine-releasing activity when similarly made to react with normal rat mast cells. The further study of such a structure should throw new light on the molecular basis of allergen-antibody triggering of mast cells.


Soluble proteins in the human atherosclerotic plaque. With spectral reference to immunoglobulins, C3-complement component, alpha 1-antitrypsin and alpha 2-macroglobulin. A number of soluble proteins contained in human aortic intimal tissue was extracted into buffered saline (pH 7.4) and identified and quantitated by immunoelectrophoresis and immunodiffusion. The proteins included IgA, IgG, IgM, B1C (C3), alpha 1-antitrypsin, alpha 2-macroglobulin, fibrinogen, albumin, LDL, HDL, alpha 1-acid glycoprotein, beta 2-glycoprotein, transferrin and ceruloplasmin. The concentration of soluble proteins was significantly higher in the atherosclerotic intima than in the normal intima. The diseased intima also contained a small amount of tissue-bound IgG, IgA and B1C which was extractable with citrate buffer at pH 3.2. The vascular band IgG, and B1C were shown by enzymatic and immunohistochemical studies to be closely associated with the collagenous tissue of the plaque. The Ig contained in the atherosclerotic plaque may be derived in part from the biosynthesis of Ig by the artery, since the incorporation of 14C-labeled leucine into IgG by the atheromatous plaque was demonstrable by radioimmunoelectrophoresis. In contrast to the diseased artery, the normal artery did not synthesize IgG and did not contain vascular bound IgG or complement. However, the normal artery was capable of fixing IgG and B1C eluted from the diseased artery. The present studies suggested that the IgG contained and synthesized by the plaque might represent an immune response to an endogenous or exogenous antigen closely associated with plaque collagen. IgG and B1C either alone or in the form of an immune complex also may play an important role in phagocytosis in the plaque and thereby influence the course of atherosclerosis. The proteolytic inhibitors, alpha 1-antitrypsin and alpha 2-macroglobulin, found in relatively high concentrations in the plaque, could enhance fibrosis of the lesion because of thier known inhibitory effects on collagenase and elastase.


[Counterimmunoelectrophoresis on cellulose acetate membrane with a commercial lyophilized antigen in diagnosis of human hydatidosis]. A very simplified method of crossed over electrophoresis (CIEP) was employed with a lyophilized commercially produced antigen (previously submitted to several freezing and thawing) and cellulose acetate membrane, for the diagnosis of human hydatidosis. Results are as follows: active hydatidosis (surgically confirmed): number of sera tested 35; positive 32 (91%). Sera of patients with parasitic and non parasitic illnesses (especially malignancies of liver and lung) did not show any precipitin lines. The pattern of immunoprecipitation is characteristically in the form of an "upper lip" or a thick and undulating "streak". The very thin arcs of precipitation which are occurring in some control sera, are to be considered negative. These results indicate that with regard to the rule of the "three S" (specificity, sensitivity and simplicity) the method appears sufficiently satisfactory.


An improved immunoelectrophoretic methodology for the evaluation of protein caloric malnutrition. An improved technique of two dimensional immunoelectrophoresis of serum proteins on gelatinized cellulose acetate for routine evaluation of protein nutritional state is described. The method, which allows the estimation of more than 30 protein fractions, proves extremely useful in monitoring the effectiveness of total parenteral nutrition and protein metabolic conditions. Sera samples from healthy subjects and cachectical neoplastic patients have been examined and preliminary results are reported.


Separation of human leukocyte interferon components by concanavalin A-agarose affinity chromatography and their characterization. Human leukocyte interferon (HL-IF), produced by mixed leukocytes infected with Newcastle disease virus, was resolved into three distinct fractions when chromatographed on concanavalin A-agarose. The major portion (70--75%) of interferon appeared in the breakthrough (BT fraction). The bound interferon (25--30%) was displaced from the column as two peaks: the first was eluted with 0.01 M methyl alpha-D-mannoside, yielding 15-20% of the interferon activity (alpha-MM fraction), and the second by including ethylene glycol (70%) in the eluant, yielding the remaining 5--15% of the interferon (EG fraction). No interferon was retained when HL-IF produced in the presence of glycosylation inhibitors (tunicamycin or 2-deoxy-D-glucose) was chromatographed on concanavalin A-agarose, suggesting that the fraction of interferon retained by this lectin is glycosylated. The three fractions of interferon (BT, alpha-MM, and EG) were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, cross-species antiviral activity, and neutralization by specific antisera. The BT fraction contains exclusively the 16 000 molecular weight component of human leukocyte interferon. The majority of the alpha-MM fraction (90%) is the 21 000 molecular weight component. However, the EG fraction contains the 16 000 and 21 000--23 000 molecular weight components in essentially equal proportions. On the basis of cross-species antiviral activity and neutralization by specific antisera, the BT and alpha-MM fractions are leukocyte-type interferon and the EG fraction seems to be primarily of fibroblast type.


Utilization of mismatched initiator termini by avian myeloblastosis virus DNA polymerase. DNA synthesis by avian myeloblastosis virus was studied using poly(C) as template and modified oligo(dG) as primer. The addition of one noncomplementary base to the 3'-end of the primer has no important effect on synthesis. The mispaired base is incorporated into the product and the apparent Km (for primer) and the V of the reaction remain unchanged. This confirms the absence of a 3' leads to 5'-exodeoxynuclease activity using a template that is transcribed faithfully rather than one that can undergo a slippage reaction.


Glutathione thiol esterases of human red blood cells. Fractionation by gel electrophoresis and isoelectric focusing. The number and the substrate specificities of glutathione thiol esterases of human red blood cells have been investigated by gel electrophoresis and isoelectric focusing and staining methods devised for the location of these enzymes on gels. Several glutathione thiol esterase forms, both unspecific (with respect to the S-acyl group of the substrate) and specific were found. Electrophoresis on both polyacrylamide and agarose gels resolved three enzyme components with apparently similar substrate specificity. Isoelectric focusing in liquid column separated two unspecific thiol esterase components with S-lactoylglutathione (pI = 8.4) and S-propionylglutathione (pI = 8.1) as the best substrates, respectively, and two specific enzymes, S-formylglutathione hydrolase (pI = 5.2) and S-succinylglutathione hydrolase (pI = 9.0). Isoelectric focusing on polyacrylamide gel resolved nine unspecific glutathione thiol esterase bands (between pH values 7.0 and 8.4). Partially purified glyoxalase II (S-2-hydroxyacylglutathione hydrolase, EC 3.1.2.6) from erythrocytes or liver still gave three components on electrophoresis and several activity bands on gel electrofocusing. These results indicate that human red cells contain at least four separate glutathione thiol esterases. Glyoxalase II, one of these enzymes, apparently occurs in multiple forms. These were neither influenced by preptreatment of the samples with neuraminidase or thiols nor were interconvertible during the fractionations.


The use of radio-immuno-inhibition assay for the study of the y, d and w determinants of hepatitis B surface antigen. A radioimmunoassay determination of the HBs antigen subtypes is discussed, it is simple but effective technique was used in association with the use of the Austria II kit (Abbott Laboratories). This method consists of an inhibition reaction of the Austria II test, by previous incubation of the antigen to be subtyped with a monospecific antibody. With this method we were able to distinguish the y and the d antigens as well as the w1, w3, w4 determinants of hepatitis B surface antigen. We have included a frequency table of the various HBs subtypes found among donor and patient populations in Liège.


[Immunohistochemical identification of specific antigens in stained and balsam-embedded eye lens sections]. Localization of specific proteins at preliminary stained with azan and embedded in balsam sections of amphibian eye lens (Rana temporaria, Xenopus laevis) was studied by indirect immunofluorescence. The analysis was performed on the lens of intact animals as well as on lens induced in amphibia gastrula ectoderm in vitro. To remove balsam the lens sections were successively washed by xylene, ethanol and saline (pH 7.1). Then they were treated according to the general principles of immunohistochemical analysis. Specific antigens have been thus shown to be detectable at lens sections which had been kept in balsam for over a year. The immunofluorescent reaction to lens proteins was negative at more prolonged storage of the sections (2--3 years).


[Two-layer rocket-electrophoresis on cellulose-acetate membranes]. Two-floor rocket-electrophoresis on gelatinated acetate cellulose membrane "Cellogel" has been developed. The method is based on electroimmunodiffusion detection of the antigen on the acetate-cellulose membranes, containing monospecific antiserum of the test-system. The procedure is followed by the detection of precipitation bands ("rockets") by staining, if the reaction is conducted in the visible zone or by the further treatment of the acetate-cellulose strips, containing invisible precipitates with antiglobulin antibodies, the complement or their combination. An increase in the method sensitivity up to 30-60 ng/ml in the visible zone of the reaction is achieved by simultaneous reduction in the antibody concentration and the growth of the absolute quantity of the antigen, subjected to electrophoresis, up to 50-100 microliters. The method has been applied to human alpha-fetoprotein.


Increased serum prolactin pituitary reserve in patients with prostatic neoplasms. Prolactin (PRL) pituitary reserve was measured in 8 patients with prostatic adenoma and 11 patients with prostatic carcinoma. The PRL reserve was significantly higher in these patients compared to an age-matched control group of 6 subjects. A possible role for PRL-inhibiting drugs in the management of oestrogen-resistant carcinomas of the prostate is suggested.


Regulation of hormone-induced cyclic AMP response to parathyroid hormone and prostaglandin E2 in cells cultured from human giant cell tumors of bone. Cells dispersed from human giant cell tumors of bone and grown in monolayer culture increase intracellular cyclic AMP (cAMP) when incubated with parathyroid hormone (PTH) or prostaglandin E2 (PGE2). When cells are continuously exposed to PTH, cAMP levels increase acutely but then decrease rapidly to pretreatment values despite continued presence of hormone or addition of new hormone. Preincubation of cells with PTH for periods as short as 10 min results in a decrease in the capacity of cells to increase cAMP content when re-exposed to maximal stimulatory concentrations of PTH. The decrease in the magnitude of the PTH-induced cAMP response observed in cells pretreated with this hormone is dependent on the concentration of PTH present during the preincubation. The loss of cAMP response in cells pretreated with either PGE2 or PTH is hormone specific in that cells made refractory by pretreatment with one hormone still increase cAMP content when exposed to the other. Although the cells are not releasing measurable amounts of prostaglandins into the medium, pretreatment with indomethacin results in an increase in the magnitude of the cAMP response to PGE2. The PTH-induced cAMP response is not affected by indomethacin pretreatment. The loss of PTH responsiveness produced by hormone preincubation is consistent with the phenomenon of "down-regulation" observed with ligand-receptor interactions in a variety of tissues.


Continuation of fast axonal transport in regenerating axons in vitro. A previous study by McLean and co-workers reported that regenerating axons of the rabbit vagus nerve were unable to sustain axonal transport in vitro for several months after nerve injury. In contrast, we found that sensory axons of the rat sciatic nerve were able to transport 3H-labeled protein into their regenerating portions distal to the site of injury within a week after injury when placed in vitro. Transport in vitro was not significantly less than transport in axons maintained in vivo for the same period. Transport occurred in the medium that was used by the McLean group, but was significantly reduced in calcium-free medium. When axon regeneration was delared, only small amounts of activity were present in the nerve distal to the site of injury, showing that labeled protein normally present in that part of the nerve was associated with axons and was not a result of local precursor uptake by nonneural elements in the sciatic nerve. We were not able to explain the failure of McLean and co-workers to demonstrate transport in vitro in regenerating vagus nerve, but we conclude that there is no general peculiarity of growing axons that makes them unable to sustain transport in vitro.


Effects of adenosine and adenine nucleotides on synaptic transmission in the cerebral cortex. Adenosine and the adenine nucleotides have a potent depressant action on cerebral cortical neurons, including identified corticospinal cells. Other purine and pyrimidine nucleotides were either weakly depressant (inosine and guanosine derivatives) or largely inactive (xanthine, cytidine, thymidine, uridine derivatives). The 5'-triphosphates and to a lesser extent the 5'-diphosphates of all the purine and pyrimidines tested had excitant actions on cortical neurons. Adenosine transport blockers and deaminase inhibitors depressed the firing of cortical neurons and potentiated the depressant actions of adenosine and the adenine nucleotides. Methylxanthines (theophylline, caffeine, and isobutylmethylxanthine) antagonized the depressant effects of adenosine and the adenine nucleotides and enhanced the spontaneous firing rate of cerebral cortical neurons. Intracellular recordings showed that adenosine 5'-monophosphate hyperpolarizes cerebral cortical neurons and suppresses spontaneous and evoked excitatory postsynaptic potentials in the absence of any pronounced alterations in membrane resistance or of the threshold for action potential generation. It is suggested that adenosine depresses spontaneous and evoked activity by inhibiting the release of transmitter from presynaptic nerve terminals. Furthermore, the depressant effects of potentiators and excitant effects of antagonists of adenosine on neuronal firing are consistent with the hypothesis that cortical neurons are subject to control by endogenously released purines.


Effect of synthetic cyclic nucleotides on immunologic histamine release from calf granulocytes. Sensitized bovine granulocytes release histamine when exposed to specific antigens. A unique modulation of histamine release by adrenergic agents has been shown in the bovine; beta-adrenergic agonists enhance and alpha-adrenergic agonists inhibit histamine release. This is an opposite response to that reported in other species. The present study was undertaken to determine the possible relationship between cyclic nucleotides and adrenergic agents in this species. Dibutyryl cAMP enhanced antigen-induced histamine release over the complete concentration range tested (10(-6)--10(-3)M); it also overcame, in a dose-dependent manner, the inhibition of antigen-induced histamine release produced by 10(-4) M phenylephrine. The 8-bromo cGMP AND 0-MONOBUTYRYL CGMP had no significant effect on antigen-induced histamine release nor did 8-bromo-cGMP have any significant effect on the enhancement of histamine release produced by 10(-4) M dibutyryl cAMP. These findings suggest that only cAMP has a role in the modulation of antigen-induced histamine release from bovine granulocytes.


The synthesis of antigenic determinants for yeast D-mannans and a linear (1to 6)-alpha-D-gluco-D-mannan, and their protein conjugates. 2-O-Benzoyl-3,4,6-tri-O-benzyl-1-O-tosyl-D-mannopyranose and 2,3,4-tri-O-benzyl-6-O-(N-phenylcarbamoyl)-1-O-tosyl-D-glucopyranose were allowed to react with partially blocked 2-[4-(ptoluensulfonamido)phenyl]ethyl alpha-D-manno- and gluco-pyranosides. Disaccharides having alpha-D-Manp-(1 to 2)-alpha-D-Manp, alpha-D-Manp-(1 to 6)-alpha-D-Glcp, alpha-D-Manp-(1 to 6)-alpha-D-Manp, and alpha-D-Glcp-(1 to 6)-alpha-D-Manp structures, and a branched trisaccharide having the structure alpha-D-Manp-(1 to 2)-[alpha-D-Manp-(1 to 6)]-alpha-D-Manp were synthesized. The oligosaccharid:s were deblocked with sodium in liquid ammonia to give glycopyranosides having a free primary aromatic amine which were converted into isothiocyanate derivatives with thio-phosgene. The functionalized oligosaccharides were then coupled to bovine serum albumin to give protein conjugates.


Three-dimensional arrangement of sarcoplasmic reticulum in crayfish as seen by high voltage electron microscope. Sections several micro thick of single fibres from the crayfish muscle were examined by means of a high voltage electron microscope to find out the geometry of the sarcoplasmic reticulum (SR). A sufficient contrast of the SR was achieved by lead acetate histochemical method for calcium. Longitudinally oriented files of SR vesicles at the level of A bands and interruptions in otherwise continuous SR net are the most conspicuous features.


Cells of origin of the hippocampal afferent projection from the nucleus reuniens thalami. A combined Golgi-HRP study in the rat. Neurons of the nucleus reuniens thalami stained with Golgi methods are compared to cells in this nucleus labelled in retrograde fashion after hippocampal injections of horseradish peroxidase. The cellular morphology ranges from fusiform to multiangular with most cells showing radiating processes characteristic of neurons in the reticular core. Dendrites are long and relatively smooth, with a few sparsely distributed spinous processes. These cells are comparable to the cholinergic cells of the median septal/diagonal band area which also project into the hippocampal formation.


Assembly and maturation of HLA-A and HLA-B antigens in vivo. HLA-A and HLA-B antigens are integral membrane glycoproteins which consist of a glycosylated heavy chain embedded in the membrane in noncovalent association with beta 2-microglobulin, a water-soluble polypeptide. The assembly and maturation of these antigens has been studied in vivo in the human B lymphoblastoid cell line T5-1 (HLA-A1, -A2, -B8, -B27). Two antigenically distinct populations of HLA-A and -B heavy chains can be detected by antisera which recognize determinants sensitive to the conformation of the heavy chain. One heavy chain population is associated with beta 2-microglobulin, whereas the other population is not. These populations can be further distinguished by their oligosaccharide structure and their localization within the cell. Pulse-chase experiments demonstrate a precursor-product relationship between these heavy chain populations and suggest the following pathway for the assembly and maturation of HLA-A and -B antigens. The completed heavy chains initially carry high mannose oligosaccharides and are largely or wholly associated with beta 2-microglobulin. During the next 10-15 min, association with beta 2-microglobulin occurs and the heavy chain conformation is altered. Beginning at about 30 min after synthesis, the oligosaccharides are converted from the high mannose form to the complex form, and between 60 and 80 min after synthesis, the mature antigens appear at the cell surface. These observations are discussed in relation to in vivo and in vitro studies on the biosynthesis of a variety of secreted proteins and membrane proteins.


Testicular germ cell tumors: diagnosis, management, and the potential for cure. Even in advanced stages, testicular germ cell tumors are now curable. Pathologic classification proposed by W.H.O. affords more precise histologic definition; nevertheless, an improved staging classification more relevant to combination chemotherapy is required. Diagnostic evaluation should include the newer serum protein markers, lymphangiography as well as ultrasound and/or computerized axial tomography when clinically indicated. Combination chemotherapy drug regimens have made a major impact on survival of patients with testicular germ cell tumors because they can induce a high percentage of durable complete remissions. Only prospective trials can define the optimum number of drugs needed and the length of time of treatment to obtain cure. The role of combined modality therapy (radiation, reductive surgery, chemotherapy) needs to be further defined. It is hoped that ongoing clinical trials will answer many of these important questions.


Phase I and II clinical study of anhydro-ara-5-fluorocytosine (AAFC) and ICRF-159 combination in adenocarcinoma of digestive origin. The results of a phase I--II study of a combination chemotherapy with AAFC and ICRF-159 in advanced adenocarcinoma of digestive origin are presented. Myelosuppression was the dose-limiting toxicity with anemia, leukopenia, and thrombocytopenia. The maximum tolerated dose of AAFC in the combination program was 650 mg/m2 I.V. weekly. ICRF-159 was given in a 3-day course every 3 weeks and the dose was escalated from 125 mg/m2 to 500 mg/m2 daily. Bone marrow toxicity was noticied at the first escalation level and all dose levels were similarly toxic. The results of this combination chemotherapy were: two partial responses in 14 patients with gastric cancer; no responses in nine patients with colorectal cancer; no responses in three patients with pancreatic cancer; and no responses in two patients with biliary tree cancer. In conclusion, AAFC and ICRF-159 combination chemotherapy demonstrated a low level of activity in advanced carcinoma of digestive origin. The peculiar hematologic toxicity found at the low-level dose requires further documentation and could make this drug association suitable for a phase II study in leukemia and/or lymphoma.


The biologic basis for combined modality treatment of cancer. Strategies related to cancer should be aimed at optimization of local and regional treatment. Delineation of subsets of patients with high risk for recurrence would have a significant impact, upon which such adjunctive systems of therapeutic management should be evaluated. In disseminated disease, or where the probability of dissemination is high, evaluation of any realistic potential combination of treatment for tumor control should be pursued. It is necessary to delineate optimal palliative therapy where tumor control is not possible, but also to test new drugs, immunotherapeutic tools and combination regimens in all such situations where the disseminated compartment of the disease is of high probability. Only under such circumstances will the most practical and important optimization of treatment programs be achieved. The development of the above basic attributes for each tumor type needs to be studied so that protocols can be developed with a clear understanding of the nature of the disease, condition to be studied, and the general therapeutic strategies to be followed.


[Stimulating determinants of the secondary mixed lymphocyte reaction (MLR-II)]. An anti-Ia immune serum (A . TL anti-A . TH) directed to the antigens of the stimulating cell, blocked the MLR-II. A significant correlation was observed between the anti-Ia reactivity of the immune serum, studied against a panel of eleven original H-2 haplotypes and the reactivity against the same panel of the in vitro primed responding cells (A . TL anti-A . TH). These results confirm the hypothesis that Ia antigens are the structures stimulating the MLR-II.


[Comparison of soluble antigens of 2 cultured clones, of different age, from the same strain of a flagellate parasite: Trichomonas vaginalis Donné, 1837]. Antigenic analysis released indications that there seemed to be a qualitative difference between two clones (A2, A3) as well as between a young clone A2 and an old one (the latter having been in culture for 12 months) both of them originating from the same initial clone. Such differences are reduced to quantitative level by the results of cross absorptions. Antigenic variations mentioned by the previous authors are to be questioned.


New concepts concerning the clinical control of IgE synthesis. Prospects for new therapeutic approaches to IgE-mediated allergic diseases have arisen from (1) recent experimental observations that have suggested new concepts concerning the pathogenesis of the allergic phenotype, and (2) adaptation of previously described methods for induction of specific immunological tolerance to problems to allergy. The bases for these new approaches to allergic diseases are summarized herein.


Metabolism of intravenously administered cholecalciferol in man. Following intravenous injection of (3H) cholecalciferol into healthy subjects the disappearance of label from the plasma was followed by the reappearance ("rebound") of (3H) radioactivity associated exclusively with cholecalciferol. Lipoprotein fractionation of plasma revealed an increasing association of label with protein rather than lipoprotein during the rebound phase. We conclude that the rebound of plasma radioactivity reflects the transfer of label from lipoprotein to Vitamin D-binding globulin in the liver followed by its release into plasma.


Sister chromatid differential staining by direct staining in Na2HPO4-Giemsa solution and the mechanism involved. The direct staining of BrdU-substituted Chinese hamster chromosomes in a Na2HPO4-Giemsa solution without any pretreatments resulted in a B-dark type SCD istituted (TB) chromatids stained light. Detailed examinations of the staining process suggested that the Na2HPO4 solution acts to collapse chromosomes whereas the Giemsa dye works to reconstruct the collapsed chromosomes, and that during the reconstruction process preferential binding of the Giemsa dye to the BB-chromatids occurs to produce the B-dark SCD. It was revealed that not only the time but the temperature at which chromosome preparations are kept prior to use considerably affect the occurrence of SCD.


[Primary retroperitoneal tumours (author's transl)]. Mean duration from symptoms to diagnosis in 87 patients with primary retroperitoneal tumour was 4.8 months for malignant tumour (67 patients), 18.1 months for benign tumour (20). The cardinal symptoms were palpable abdominal mass (68%), feeling of fullness and nausea (64%), as well as ill-defined abdominal pain (55%). Radiological diagnosis, especially selective angiography, provided important pointers to the global diagnosis of "retroperitoneal tumour". Small-needle biopsy, guided by ultrasound, has now become available in the pre-operative diagnosis of the type and malignancy of these tumours. After radical resection of 30 malignant tumours there was a mean survival time of, so far, 30.2 months. Results were worse with palliative malignant-tumour removal in 25 patients, at an average of 17.3 months. Survival time was only 4.8 months for 29 patients declared inoperable. In individual instances cytostatic treatment achieved an increase in survival time of up to 12 years.


[Anti-arrhythmic effect of tocainide (lidocaine congener) on ventricular arrhythmias (author's transl)]. The antiarrhythmic effect of tocainide, an amine analogue of lidocaine, was tested in the course of a double-blind trial on 12 patients with ventricular arrhythmias. Daily dose was 400 or 600 mg every eight hours. In all tested patients the frequency of ventricular extrasystoles, analysed by means of ambulatory 24-hour ECG monitoring, decreased by an average of 70%. In four of nine patients the severity of the ventricular extrasystoles (Lown's classification) was improved by at least one functional class. There was no correlation between the anti-arrhythmic effect and the plasma level of tocainide, which was between 4.1 and 9.7 micrograms/ml. Four patients had side effects in the form of CNS symptoms, but the drug did not have to be discontinued. Tocainide is an orally effective antiarrhythmic drug, an alternative in the treatment of ventricular arrhythmias.


Binding of bovine immunoglobulins to pituitary intermediate lobe cells. Basophilic granular cells of the bovine pituitary intermediate lobe and intraglandular colloid show an affinity for immunoglobulin IgG, IgA, and IgM. The cells were identified by direct and indirect fluorescent staining methods using blocking tests as controls. It is suggested that these cells may play some role in the synthesis and release of immunoglobulins or their determinants to the intact Ig molecule.


Activity staining for glutamate and alanine dehydrogenases in polyacrylamide gels of varying porosity. A study of bovine placental enzyme forms. A procedure has been developed for characterizing the various molecular forms of placental and liver glutamate dehydrogenases through a combination of activity staining and varying gel pore size in electrophoresis. At a concentration of 2 mg/ml, the bovine liver GDH remained associated in a very high molecular weight form, while the placental enzyme was substantially dissociated to a molecular species of near 240,000 molecular weight and several charge isomeric species of near 160,000 molecular weight. The general approach outlined here provides a means of definitely correlating the electrophoretic behavior of various dehydrogenase isozymes with both glutamate and alanine dehydrogenase activities and molecular size and should be applicable, even in crude extracts to other dehydrogenase enzymes which exhibit multiple forms or states of association.


Neoplasm localization with radionuclides. Tumor scintigraphic localization of neoplasms can be done in two ways: indirectly and directly. The first method shows alternations of the normal structure of the organ, such as "cold lesions" in liver and thyroid. Abnormalities in function as increased permeability of the blood barrier results from abnormal deposition of the radionuclide in the brain scintigram of a patient with neoplasm. Increased focal areas of uptake of bone-seeking radionuclides are very characteristic of metastases. The direct methods depend on preferential uptake of the radionuclide by the neoplastic tissue resulting from altered metabolism (e.g. Se-75). Other agents such as Gallium-67 have affinity for neoplasms. Another approach is to use antineoplastic agents and radioactive antibodies which will localize in the tumor. At this stage the most useful neoplasm seeking agents are Gallium-67 citrate and 111In-Bleomycin, even though infections can give false positives. The possibility should be considered of enhancing the uptake of radionuclides by neoplastic cells using increased O2 concentration.


The effect of dose loading and of double labelling with 57Co and 125I on the tissue distribution in animals. Dose loading effects upon the performance of 57Co-bleomycin as a tumor localizing agent have been investigated in Rhabdomyosarcoma bearing Wag/Ry rats. The addition of non-radioactively labelled Co-bleomycin increased the relative uptake of 57Co-bleomycin in rapid growing tumors, but the addition of non-chelated bleomycin had no influence at all. In our experimental system, iodinated bleomycin generally labelled by reaction with ICl, was found to be an unsatisfactory tumor localizing agent. In order to combine the useful localizing properties of Co-bleomycin with the qualified detection properties of some iodine isotopes, we attempted to prepare bleomycin doubly labelled with Co and I. However, we were unable to prepare 57Co-125I-bleomycin by general labelling with ICl. This result indicates that both labels need the imidazole ring for the formation of a stable, labelled bleomycin.


HLA B27 and defects in the T-cell system in Whipple's disease. The cellular immune system was tested in nine patients with Whipples' disease. Three patients had active disease, and six had been in remission for up to 10 years. Intradermal delayed hypersensitivity reactions to candidin, trichophytin, tuberculin and varidase, T-cell counts as determined by E-rosettes, allogeneic stimulation of lymphocytes in the mixed lymphocyte culture, and mitogenic activation of lymphocytes by concanavalin A, phytohaemagglutinin and by pokeweed mitogen, were tested in the patients and compared with control subjects. HLA typing was performed in all patients. The reaction to tuberculin and varidase, the T-cell counts and the activation of lymphocytes by concanavalin A were significantly reduced in patients with active disease and in patients during remission. The reaction to candidin and trichophytin was poor even in the controls. The mean results of the mixed lymphocyte culture, phytohaemagglutinin, and pokeweed mitogen activation tests were not significantly different from the controls. In patients with active disease the mixed lymphocyte culture reaction and the T-cell counts were less than in patients in remission. The results suggest a persistent defect of T-cells in patients with Whipple's disease, a defect that is more severe in patients with active disease. The finding of HLA B27 in four of thenine patients supports the hypothesis of primary rather than secondary impairment of the cellular immune system in Whipple's disease.


Clinical experience of encainide (MJ 9067): a new anti-arrhythmic drug. Encainide is a new anti-arrhythmic drug, which is highly effective against ventricular extrasystoles, both single and coupled, in the dose range of 80--140 mg i.v. Ventricular extrasystoles were abolished in 31 out of 33 cases treated. The drug is also relatively effective against supraventricular extrasystoles, but has little effect on atrial fibrillation. Both subjects with clinically normal hearts and those with ischaemic heart disease have been successfully treated. The drug prolongs the QRS and QT duration. Its effectiveness appears to be of the same order of magnitude and the range of indications similar to those of aprindine and lorcainide. Further study of the drug seems to be warranted.


Immunological activity of a T hybrid line. I. Production of an H-2-related suppressor factor with specificity for sheep red blood cells. T lymphocyte hybrid lines have been produced by fusion of the thymoma BW 5147 with spleen cells of C57BL/10 mice primed to sheep red blood cells (SRBC). The supernatant (culture fluid) of a T hybrid designated A 1 was able to suppress the primary (IgM) and secondary (IgM and IgG) antibody responses to SRBC in vitro. The suppressive activity of supernatants could be titrated to 50% end points at final dilutions of up to 1 : 270. The suppression affected only SRBC and haptens coupled to SRBC, except when used at high concentration when some nonspecific suppression was observed. Absorption of the A 1 supernatant with SRBC removed all activity, while several other species of red cells failed to do so. The suppressor factor present in A 1 supernatant was not removed by anti-Ig adsorbents, but was removed by anti-H-2 antibodies reacting specifically with the haplotype of the spleen cells used in the fusion (H-2b). The cellular target of action of the factor was apparently a B cell, based on absorption with different cell populations. No genetic restrictions in the activity of the factor were found. A 1 cells carried H-2 and Thy-1 alleles of both parental cells and formed rosettes with SRBC.


Epidermal growth factor stimulates proliferation of rat hepatoma cells producing alpha-fetoprotein. Epidermal growth factor stimulated both [3H]thymidine uptake and proliferation of rat AH66 hepatoma cells. However, the increase in cell number was not accompanied by a proportional increase in the levels of alpha-fetoprotein of the culture media. The effects of EGF on the cell proliferation were antagonized by N6,O2'-dibutyryl cAMP.


Effect of hyperthermia on DNA single-strand breaks induced by bleomycin in Hela cells. The effect of hyperthermia (43 approximately 45 degrees) on DNA single-strand breaks induced by bleomycin was examined in HeLa cells by the alkaline sucrose gradient sedimentation analysis. When HeLa cells were exposed to a temperature of 43 degrees approximately 45 degrees for 1 hr, only a small number of DNA single-strand breaks than bleomycin treatment at 37 degrees. The DNA strand breaks induced by bleomycin were almost completely repaired by post-treatment incubation of the cells in the control medium at 37 degrees. On the contrary, the repairing capacity of the cells was markedly inhibited by post-treatment incubation at 43 degrees, and this inhibition of cell repair seemed irreversible because no repair was found after an additional incubation of the cells at 37 degrees for 1 hr. These results support the possibility that the mechanism of synergism between the effect of bleomycin and hyperthermia may be related to the inhibition of DNA repair.


[Prenatal recognition of omphalocele (author's transl)]. The prenatal diagnosis of omphalocele is reported. The characteristic signs in the ultra-sound scan were a small trunk, an angulation of the spine and mass adjacent to the absomen of the fetus. By x-ray amniography the ultra-sound findings were confirmed at further abnormalities outlined. The alpha fetal protein levels in the maternal serum and in the amniotic fluid are markedly elevated.


Site specificity of bleomycin-mediated single-strand scissions and alkali-labile damage in duplex DNA. Form II PM2 DNA, which contained bleomycin-mediated single-strand breaks, was purified and treated with the extracellular endonuclease from Alteromonas BAL 31. This enzyme cleaves the phosphodiester backbone opposite a single-strand break to yield a double-strand break. The locations of these double-strand breaks were determined relative to the cleavage sites produced by the restriction enzyme HindIII. The experimental procedure was as follows. Form I PM2 DNA was treated with bleomycin to produce alkali-labile bonds. These were hydrolyzed by alkali treatment and the DNA, now containing single-strand breaks, was purified and treated with the BAL 31 enzyme and the HindIII enzyme to determine the positions of the original alkali-labile bonds. It was found that the single-strand breaks and alkali-labile bonds were introduced at preferred sites on the PM2 genome, since electrophoretic analyses of the DNA after the HindIII digestion revealed DNA bands of discrete sizes. The molecular weights of the DNA fragments produced by these treatments indicate that single-strand breaks and alkali-labile bonds occur at the same sites as those previously determined for direct double-strand scissions introduced by bleomycin at neutral pH. Some of the specific sites of double-strand scissions mediated by bleomycin at neutral pH (Lloyd et al., 1978b) are also shown here to be relatively more reactive than other sites when the DNA contains superhelical turns.


Bleomycin fragmentation of duplex DNA occurs as staggered single-strand scissions. Electron microscopy of purified full-length linear duplex molecules produced by bleomycin reaction with PM2 DNA revealed low frequencies of closed circular duplex molecules as well as linear duplex molecules with opposed ends (cyclized molecules which have dissociated to yield a gap between the termini). The occurrence of these latter forms indicates that double-strand scissions produced by bleomycin reaction consist of two single-strand scissions which are physically staggered on the complementary strands. Analysis of the temperature dependence for cyclization led to the estimate that an average of 1.7 +/- 0.44 base-pairs (2.6 +/- 0.5 base pairs without base-stacking energies) occur between the staggered breaks. The reassociated termini cannot be ligated with T4 ligase. When PM2 DNA was fragmented at several sites within each molecule, circular duplexes and linear duplexes with opposed ends with a range of sizes from 350 base pairs up to full-length PM2 DNA were observed. Analysis of the frequency distribution of lengths of these fragments indicates that most, if not all, of the specific sites for bleomycin-directed double-strand scissions in PM2 DNA contain representatives of the same two base single-stranded termini.


[Treatment of prostatic neoplasms].

[Treatment of prostatic neoplasms]. Early stage carcinoma of the prostate is curable. Not every patient must be treated because there are significant differences in the biologic activity of the tumors. This causes a sharp difference between morbidity and mortality rate in prostatic cancer. Besides the clinical staging morphological malignancy grading is the trend factor for the treatment that has to be chosen because it has shown to be a reasonable and feasable prognostic indicator.


[Medical management of tachycardias (author's transl)]. Guidelines for a step-wise plan of treatment of tachycardias have been compiled based on clinical empirical experience and with the aid of surface electrocardiograms, intracardial electrograms and stimulation techniques. The plan is primarily with the aid of surface electrocardiograms, intracardial electrograms and stimulation techniques. The plan is primarily oriented with respect to the antiarrhythmic efficacy, the adverse reactions and the practicability of the respective agents. Any type of tachycardia, including premature atrial or ventricular contractions, may be regarded as indication for treatment. Treatment is not indicated only in those asymptomatic patients with rare and evanescent tachycardias and in those with less than 300 premature contractions per hour. Beta-adrenergic blockers are the drugs of choice for the persistent sinus tachycardia. Should the latter agents be contraindicated, propafenon, amiodarone or aprindine may be administered. Verapamil and/or digitalis are indicated only for suppression of paroxysmal sinus tachycardias. Atrial premature contractions are best managed with guinidine or disopyramid. An acute reduction of rapid ventricular rates associated with atrial tachycardias, atrial flutter or fibrillation can best be attained through the administration of verapamil prior to digitalis or beta-adrenergic blockers. Re-establishment of sinus rhythm and prophylactic suppression of the latter should be undertaken with quinidine or disopyramid in combination with digitalis and/or either a beta-adrenergic blocker or intravenously-administered verapamil. Verapamil is the drug of choice for initial management of AV-junctional tachycardia for which a combination with digitalis may be considered. An alternative combination is that of a beta-adrenergic blocker and digitalis. For the acute treatment of ventricular tachycardias, lidocain has proved most effective. Although ajmaline and/or propafenon may be given should no response be obtained, electrical cardioversion would be more appropriate. To prevent ventricular tachycardia or when treatment is indicated for ventricular premature beats, ajmaline, propafenon, quinidine, disopyramid or mexiletine, occasionally in combination with a beta-adrenergic blocker should be employed. Verapamil and/or ajmaline, are usually very effective for termination of reciprocal tachycardias. Ajmaline or propafenon in combination with a beta-adrenergic blocker is recommended for the prophylactic treatment of reciprocal tachycardia. In patients who additionally have bradycardia, prolonged QT-intervals or pre-excitation syndromes, the guidelines should be modified accordingly.


Immunological properties of peptic fragments of bovine serum albumin. The effect of peptic degradation on the immunological and antigenic properties of bovine serum albumin (BSA) was investigated. Molecular fragments obtained after various times of digestion (3-360 min) were studied. Enzymatic digestion resulted in a rapid loss of serologically defined antigenic determinants. The immunogenicity of the fragments as measured by the level of reaginic and total anti-BSA antibody response in BDF1 mice was also diminished. Pre-treatment of mice with fragments exhibiting a low density of B-cell interacting determinants before immunization with BSA, resulted in significant suppression of both the primary and secondary antibody response. The most effective immunosuppressive fragments were obtained following removal of peptides which bound to anti-BSA antibodies. The suppression was shown to be antigen specific. It was concluded that separate determinants on the BSA molecule were responsible for the immunogenic and suppressive properties of the antigen.


B-cell subsets responsive to fluorescein-conjugated antigens. III. Differential effect of E. Coli lipopolysaccharide on T-dependent and T-independent responses in vivo. The effect of E. coli lipopolysaccharide (LPS) on the induction of both hapten-specific immunity and tolerance was studied in an in vivo system utilizing putative T-cell dependent (TD) or T-cell independent (TI) challenge antigens. The administration of LPS 1 day prior to challenge preempted the response of C3D2 mice to a TD antigen (FL-KLH) but had little effect on the response to FL-Ficoll, a TI antigen. LPS did not affect the responsiveness of C3H/HeJ mice, an LPS-unresponsive strain, to either antigen. The reduction of the response to a putative T-dependent antigen by LPS pre-treatment was only temporary since mice challenged 7 days after LPS responded normally in vivo. We also confirmed that LPS administered shortly after a tolerogen prevented FL-specific IgG tolerence induction and produced B-cell priming to a subsequent T-dependent antigenic challenge. LPS, however, did not significantly interfere with tolerance induction in terms of the IgM responce to either challenge antigen. These results suggest that LPS acts either directly or indirectly on a subpopulation of B cells responsive to a TD antigen. Our data further reflect the heterogeneity of B-cell subpopulations responsive to various polyclonal activators.


Inhibition of antigen-induced lymph node cell proliferation by murine amniotic fluid and its components. Murine amniotic fluid (MAF), alpha-foeto-protein (AFP) and MAF depleted of AFP by affinity chromatography (MAF-AFP) inhibited the T-cell dependent in vitro proliferative responses of lymph node cells sensitized to a variety of soluble antigens. Variable degrees of inhibition were observed with the different antigens used in the assay. In general, the higher the proliferative response induced by a particular antigen, the less it was inhibited by the three inhibitors. Enhancement of proliferation was not infrequently observed at lower concentrations followed by a dose-dependent inhibition as the concentration of the inhibitor was increased. Usually the order of inhibition was MAF greater than MAF-AFP greater than or equal to AFP although variations in inhibitory potency were noted between different preparations of AFP and MAF-AFP. The existence of inhibitors in preparations of MAF depleted of AFP raised the question as to whether MAF contains single or multiple inhibitory factors. The most facile explanation is that two inhbitors exist; AFP and the as yet uncharacterized non-AFP suppressor present in MAF-AFP.


Antigenic differences between alveolar and peritoneal macrophages of the rat. Lack of population-specific determinants. Antisera produced in rabbits against adherent cells of rat alveolar or peritoneal lavage fluid (anti-rat alveolar macrophage sera, ARAMS, or anti-rat peritoneal macrophage sera, ARPMS) were used to detect antigenic differences between alveolar (AM) and peritoneal (PM) macrophages in an indirect membrane immunofluorescence (IMF) test. Of all sera tested, the IMF titres were higher with cells of that population which was used for immunization. These differences were found before and after exhaustive absorptions with insolubilized rat plasma, rat erythrocytes, and non-adherent rat kidney, spleen, thymus and bone marrow cells. The differences were not due to antigens specific for one of the macrophage populations, as demonstrated by cross-adsorption studies with macrophages of different localization. It is assumed that two or more macrophage specific antigenic determinants are present in different density in the two macrophage populations.


Preferential induction of memory T cells for delayed-type hypersensitivity with reduced and alkylated human serum albumin in mice. Reduction and alkylation of human serum albumin (HSA) resulted in molecular aggregation of the protein. The reduced and alkylated antigen (RA-HSA) was lacking in the ability to induce delayed-type hypersensitivity (DTH) response as well as antibody response to native HSA in mice, although native HSA induced both responses. On the other hand, RA-HSA could stimulate a priming function that accelerated and enhanced the DTH response to native HSA, which, however, failed to stimulate the function. Thus, DTH-related memory activity was dissociated from DTH-related effector activity. The DTH-related memory activity, manifested by the accelerated an enhanced response in RA-HSA-primed mice, could be transferred antigen-specifically by their spleen cells and T-cell enriched fraction, but not by their T-cell depleted spleen cells. The RA-HSA-primed T cells, however, failed to transfer the effector function for DTH response. These results suggested that DTH-related memory T cells belong to different subset(s) of T cells from effector T cells for a DTH response.


Delayed hypersensitivity to Staphylococcus aureus in mice: in vivo responses to isolated Staphylococcal antigens. The development of delayed hypersensitivity (DH) to Staphylococcus aureus in Swiss mice was evaluated by the footpad (FP) assay. In order to determine which component of the bacteria was responsible for the in vivo immune reactivity, purified Staphylococcal cell wall, cell membrane, protein A, lipoteichoic acid, teichoic acid, as well as lipid-free membrane proteins were isolated. The immune responses of mice receiving one to eight S. aureus injections indicated that the first DH peak, following three injections, was primarily dependent upon protein antigens associated with the bacterial membrane. Increased bacterial injections gave rise to a second DH peak following seven injections which was dependent upon multiple bacterial components including cell wall, protein A, and membrane proteins.


Normal immunosuppressive protein: in vitro inhibition of DNA synthesis in T and B lymphocytes and lymphoid cell lines. The biologic activity of normal immunosuppressive protein (NIP) isolated from human plasma was studied. NIP was found to inhibit the proliferation of both T and B lympohcytes in vitro. It suppressed the DNA synthesis of normal mouse lymphocytes responding to the mitogens phytohemagglutinin and lipopolysaccharide, as well as the [3H]Thymidine and [3H]leucine uptake by T and B lymphoid cell lines of human and murine origin. The lymphoid specificity of NIP was demonstrated by showing that DNA and protein synthesis of normal and transformed fibroblasts and other nonlympohid cell lines was not affected by NIP treatment. Furthermore, by using lymphoid cell lines we were able to show that 1) NIP inhibits the process of ongoing DNA and protein synthesis; 2) the duration of the cells' exposure to NIP is crucial for obtaining optimal effect; and 3) the inhibitory effect of NIP is totally reversible.


[New studies on the pathogenesis of lupus erythematosus]. The pathogenesis of the lupus erythematosus results from co-operation of three principles: (1) genetical disposition, (2) increased reactivity of the immune system, (3) different exogenic influences. The genetical disposition is confirmed by family investigations, metabolic disorders and immune anomalies as well as by parallels to animal models. The reaction manner of the immune system is genetically determined. Exogenic factors influence the immune system (behaviour) either as starter or by modifying the genetical material. The most striking humoral immune phenomenon is an immense number of (auto-)antibodies. Investigations of xeroderma pigmentosum as well as with DNA of different antigenity have shown that an increase of the antigenicity is unlikely for this phenomenon. In the serum of patients there were established and partially characterized factors (mitogens, granulocytic adherence-factor) for increasing the immune reactivity. The increase of such factors may be the sequence of a T-suppressor cell-defect. Like-wise no interdigitating cells in systemic lupus erythematosus have been found, cells which may be responsible for the terminal differentiation of T-lymphocytes. The mode of action of exogenic factors is represented and discussed with the example of the UV provocation.


Simultaneous determination of the amounts of metallic and "reducible" silver in histologic specimens. Acids and weak complexing agents (pK less than 8) are not able to remove, without leaving a residue, silver bound to biological tissues by ionic or complex bonds ("reducible" silver), whereas, strong complexing agents (pK greater than 8) can also partially or completely dissolve metallic silver formed under the influence of reducing groups in the tissue. For this reason, the chemical nature of the silver contained in tissue sections, be it metallic or reducible, must not be determined on the basis of solubility tests; moreover, the amount of neither of the two above fractions can be determined by removing the other with any kind of washing. Using radioactive impregnating baths, radioactive silver bound to the tissue as reducible silver can be replaced in a quantitative manner with inactive silver ions by means of a one-hour incubation in 1% inactive silver nitrate dissolved in 10% acetic acid, but the radioactive silver existing in reduced (atomic) state will be left unaffected. Consequently, radioactivity remaining in the tissue after the above treatment represents metallic silver. The amount of reducible silver can be calculated by subtracting that of the metallic silver from the total silver content of the sections.


Kinetics of formation of metallic silver and binding of silver ions by tissue components. The effect of time on the formation of metallic silver by tissue reducing groups follows a curve which can be divided into three main parts. In the first, which may last for several hours, the reaction is very slow, and only an undetectably small amount of metallic silver is produced. In the second period the speed of the reaction first increases in a progressive manner and then begins to decrease gradually; during the third period the speed approaches zero asymptotically. Binding of the silver ions by the tissue commences initially at its fastest rate; the level then decreases steadily to zero within about a quarter of an hour. There is no direct relationship between the amount of silver ion bound to the tissue and the formation of metallic silver. The latter cannot take place by way of direct (non-catalysed) reaction. The following mechanism is proposed for the process: Transfer of electrons from the reducing molecules to the silver ions is mediated at first by certain tissue sites (catalytic points) and then also by the steadily increasing total surface area of the metallic silver grains (autocatalysis). On the basis of this mechanism, several anomalies of both the argentaffin and argyrophil reactions are explained.


Factors affecting the formation of metallic silver and the binding of silver ions by tissue components. The rate of formation of metallic silver has a maximum when plotted as a function of pH. The site of this maximum on a pH scale differs noticeably for various tissue elements. By contrast, the amount of silver ions bound to the tissue is a monotonously increasing function of the pH. A temperature rise decreases the length of the induction period and increases the gradient of the ascending section of the kinetic curve representing the formation of metallic silver. It also increases the maximum amount of silver ions bound to the tissue. An increase in the concentration (activity) of the silver ions in the impregnating bath has the same effect. Chemical composition and concentration of the complexing agent, as well as "special" ions in the impregnating bath to which earlier some definitive role has been attributed in the silver staining methods, proved to be ineffective when both pH and activity of silver ions were kept constant. Illumination of the reaction was also ineffective. The kinetic curves obtained in nonaqueous but polar media (e.g., acetone) exhibited the same qualitative characteristics as those obtained in aqueous solutions. No reaction between silver ions and tissue was observed in apolar solvents.


The histochemistry of complex carbohydrates in the testes of patients of idiopathic male infertility. In testis tissues from patients with idiopathic male infertility, complex carbohydrates have been studied by means of light microscopic histochemical methods, comparisons being made with the normal testes. In the infertile testes, histochemical reactions for acidic and neutral complex carbohydrates were in general weaker in intensity than in the normal testes. In the seminiferous tubules and interstitium of the infertile testes, a histochemical finding of primary importance was obtained with the demonstration of galactose deficiency in the complex carbohydrates involved. These glycoproteins contained only small numbers of sulfhydryl and disulfide groups. The possible histophysiological significance of these complex carbohydrates is discussed, with special reference to their histochemical properties determined in the present study.


Quantitative succinate-dehydrogenase histochemistry. I. A Methodological study on mammalian and fish muscle. In enzyme histochemistry formazan production can be used as a measure for oxidative enzyme activity. The formazan deposits can be measured quantitatively per cell with a scanning and integrating microspectrophotometer. Optimal conditions are described for the estimation of histochemical succinate dehydrogenase activity in sections of fish bodymusculature and mouse soleus and plantaris muscle. It is shown that when proper measuring conditions are chosen a ditetrazolium salt (TNBT) can be used in quantitative enzyme histochemistry and that the optimal conditions for the histochemical succinate dehydrogenase reaction in muscle fibres of fish and mouse muscle are somewhat different for these two species. The differences in pH, temperature and succinate sensitivity are the most prominent.


Quantitative succinate-dehydrogenase histochemistry. II. A comparison between visual and quantitative msucle fibre typing. Most muscles exhibit a mosaic pattern of staining intensities of their muscle fibres after the histochemical reaction for succinate dehydrogenase (SDH). Visually these muscle fibres are usually classified into three groups: with low (A-fibres), intermediate (B-fibres), and high (C-fibres) enzymatic activity (staining intensity). Cytospectrophotometric methods were employed to investigate whether discrete groups of muscle fibres could be discerned, comparable to those found after the visual classification. The classifications were based on quantitative parameters of the total absorbance per cell and the distribution of the coloured endproduct over the fibre cross area.


Quantitative succinate-dehydrogenase histochemistry. III. Variations in histochemical succinatedehydrogenase activity in different cross-sections of the same muscle fibre. The variation in histochemical SDH-activity at different levels in the same muscle fibre was determined in muscle fibre cross-sections both by visual classification and quantitative determination of the formazan-deposits. This work resulted in a confirmation of the earlier micro-biochemical studies of Spamer AND Pette (1977, 1979) and Lowrey et al. (1978) that the activity of enzymes of the citric acid cycle is not homogeneously distributed in a muscle fibre over its entire length. In addition it is shown that the observed variations in histochemical SDH-activity strongly interfere with the visual muscle fibre typing. Some of the possible causes for these variations in histochemical SDH-activity (section-thickness, presence of the motor-endplate) and the implications of these findings for the relation between histochemical characteristics and functional properties of the muscle fibres are briefly discussed.


Immunocytochemical technique for detection of prolactin (PRL) and growth hormone (GH) in hyperplastic and neoplastic lesions of dog prostate and mammary gland. An immunocytochemical technique was described to test for immunoreactive prolactin (PRL) and growth hormone (GH) in spontaneous and experimentally induced hyperplastic and neoplastic lesions of the prostate and mammary gland. The dog was used as an animal model. The specificity and validity of the immunocytochemical staining procedure and of the antisera to canine PRL and canine GH can be regarded as established for the demonstration of PRL- and GH-dependent staining respectively. In mammary and prostatic tissues, both endogenous PRL and GH as well as intracellular free binding sites (for exogenous PRL and GH) were detected immunocytochemically. The technique presented seems to be an important tool to localize putative target sites for pituitary hormones in hormone-dependent hyperplasia and neoplasia.


Histochemical reactions of gastrointestinal mucosubstances with high iron diamine after prior oxidation and methylation of tissue sections. High iron diamine reactions after the prior methylation and oxidation of tissue sections with performic acid or potassium permanganate (metox-HID or ox-met-HID) in epithelial mucosubstances and in mucosal mast cells were studied in tissue samples from the human gastrointestinal tract and were compared with reactions with high iron diamine without any pretreatment (HID) and high iron diamine with the prior methylation (met-HID). High iron diamine reactions after the prior oxidation (met-ox-HID, ox-met-Hid and ox-Hid) demonstrated mucosubstances in a way which seemed to operate by the staining of acidic groups evoked by the oxidation of the tissue sections. These acidic groups were not blocked by the methylation. It was supposed that they are sulphonic acids resulting from sulphur groups (sulphydryls or disulphides) in some mucus glycoproteins. Met-ox-HID and ox-met-HID reactions seemed to stain mucosubstances and mast cells in a similar way but differed from the ox-HID reactions with the manner which could be interpretated to be due to the blocking of free sulphate ester groups in reactions of the former. Met-ox-HID (and ox-met-HID) positive and in goblet cells of small and large bowel.


Specificity of cytochemical procedures for localising peroxidase activity in the sarcoplasmic reticulum. Cytochemical evidence is reported for substantiating the view that when lightly-fixed skeletal muscle is incubated in a diaminobenzidine-H2O2 medium at pH 5, the resulting enhanced electron opacity of the sarcoplasmic reticulum is more likely to be due to a peroxidatic activity therein rather than to a non-enzymic binding reaction. The reticulum staining is absent in incubated sections of overfixed or boiled tissue; or if hydrogen peroxide is omitted from the incubation medium; or if aminotriazole is included in the medium.


Facilitating generalization of on-task behavior through self-monitoring of academic tasks. This study (1) examined whether a self-monitoring procedure taught in a laboratory setting would increase independent on-task behavior there and would generalize without further teaching to a classroom setting, and (2) analyzed the durability of the training effects over the course of 5 months for one subject and 10 months for two other subjects. Two multiple-baseline designs, one across three normal and the other across three deviant children, showed that self-monitoring of academic task-completions facilitated on-task responding for all subjects in the generalization (classroom) setting. A subsequent reversal design showed that these effects were durable, in two of the three subjects still available, at least as much as 1 year after commencement of training. This latter design also suggested that one subject who was not maintained by self-monitoring could be supported in on-task behavior by a peer who was maintained by self-monitoring.


Electron microscopy of the rodlet layer of Neurospora crassa conidia. Neurospora crassa macroconidia possess a regularly arranged layer of small fibers (rodlets) near the spore surface. The structure and location of this layer were studied by making surface replicas, by negative staining, by freeze-fracturing and deep-etching, and by thin sectioning. When conidia were shaken vigorously in water, the layer fragmented and became separated from the surface in sheets. Negative staining of such sheets showed that the individual rodlets have a hollow central core. When conidia were shaken gently in water or fixative, large fragments of the rodlet layer often remained on the conidial surface. The fragments tended to fold back on each other such that multiple layers were sometimes seen in thin sections. It is concluded that in dry conidia the rodlets are located on the extreme outside of the spore where they form a monolayer with only occasional regions of overlap.


Immunochemical studies on the correlation between conformational changes of DNA caused by ultraviolet irradiation and manifestation of antigenicity. The conformational changes of double-stranded DNA induced during irradiation with ultraviolet, were immunologically investigated. These studies revealed that at least two distinct antigenic sites were induced in the irradiated DNA molecule, giving rise to two different antibodies specific for ultraviolet-irradiated (uv) DNA and thermally denatured DNA-like structure, and these were demonstrable using radioimmunoassay and double diffusion tests. A series of experiments, including melting profile, fluorescence intensity of the ethidium bromide complex and chromatographic behavior on hydroxyapatite, performed on the antigenically active uvDNA indicated that the duplex structure of DNA separated irregularly during irradiation. Furthermore, the data showed that the conformational determinants of uvDNA are located on the exposed single-stranded regions.


Motor neuron diseases in man and animals. Aspects of motor neuron diseases in man are described and compared with a series of animal models for these disorders. Such models provide new approaches to understanding many features of abnormalities of the central and peripheral nervous system.


Incubation of 3,4-benzo(s)pyrene with serum fractions: effect on tumor production. The effect on tumor formation in Swiss mice of incubation of 3,4-Benzo(a)pyrene [B(a)P]either with saline or saline containing serum, gamma-globulin or albumin (form mouse or rabbit) was investigated. A high incidence of sarcomas (80--100%) was obtained in Swiss mice by B(A)P incubated in vitro with rabbit serum, rabbit gamma-globulin, mouse serum, mouse gamma-globulin, and saline. A lower incidence (60--65%) was obtained with B(a)P incubated with mouse serum, mouse albumin, and rabbit albumin. The data suggest that binding of B(a)P to protein, particularly to albumin, significantly decreases the biological activity of B(a)P.


[Concentrations of cytostatic drugs in organs and tumors: comparison after intravenous and intratumoral injection (author's transl)]. The cytostatic drugs Chlorambucil and Bleomycin were labeled with 131I and with 57Co, respectively: Ruthenocenealdehyde-(N-methyl-N-beta-chlorethyl)-hydrazone and Ruthenocene-3-phenyl-prop.1-en-3-one were labeled with the gamma emitter 103Ru. In tumor-bearing mice the elimination and organ distribution of these radioactive substances were tested after intravenous (i.v.) and intratumoral (i.t.) injection of the drugs. The organ load showed lower values after i.t. than after i.v. application, while the radioactivity concentrations in the tumor were considerably increased after i.t. injection. The integrals of the concentration-time curves showed 10--80 times higher concentrations in the tumor after i.t. compared to i.v. injection of the drugs.


Analysis of Vibrio parahaemolyticus soluble antigens by employing passive hemagglutination. The passive hemagglutination assay was explored as a sensitive test of immunological reactivity of endotoxin and other antigens prepared from selected strains of Vibrio parahaemolyticus. The tannic acid procedure for passive hemagglutination, commonly used with protein antigens, was the only procedure yielding good results with V. parahaemolyticus protein extracts, endotoxins, and related preparations. These results were probably due to the presence of large amounts of protein in the V. parahaemolyticus endotoxins as determined by earlier work referenced in the text. Glucose and galactose as possible antigenic determinants in the endotoxin of a Vibrio strain were tested by inhibition tests. Cross-reactions were observed between endotoxin preparations, but were low in hemagglutination, suggesting reactions of common generic antigens. The ability of V. parahaemolyticus endotoxins to stimulate production of antibodies was determined.


A technique for estimating total spine numbers on Golgi-impregnated dendrites. The functional significance of dendritic spines and their morphological sensitivity to a wide spectrum of experimental manipulations and pathological states have led to a number of studies in which counts of dendritic spine number have been carried out. These studies have, for the most part, involved the enumeration of only those spines which protrude from the opaque shafts of Golgi-impregnated dendrites into the clear zones flanking the dendrite. Such counts, limited to only those spines which are visible, underrepresent the true total number of spines borne by the dendrites. The magnitude of underrepresentation correlates positively with dendritic shaft diameter and negatively with spine length. This seriously restricts the usefulness of comparisons of spine density between dendrites, or even between segments of the same dendrite. In the present report, a geometrically based method is presented whereby total dendritic spine numbers can be estimated with reasonable accuracy, taking into account factors such as dendrite diameter and spine length. The technique entails the following principal steps: a determination, for a given length of dendrite over which spines are to be enumerated, of the volume of the flanking zones in which spines are visible and can be counted; a determination of the volume of the entire zone which encircles the dendritic shaft and which contains all spines, both visible and not visible; and a proportional extrapolation from the number of visible spines to obtain an estimate of the true total spine number. Tests of the predictive accuracy of the technique using dendrites of known total spine number suggest that estimates which deviate from true total spine numbers by less than 10% can be achieved.


A stain for plucked anagen hairs.

A stain for plucked anagen hairs. Anagen and telogen plucked hairs may be difficult to distinguish based on their microscopic appearance. The root ends of plucked anagen hairs develop a red color when stained with 4-dimethylaminocinnamaldehyde, while those of plucked telogen hairs do not. This is the result of a reaction with citrulline-containing proteins in the internal root sheath. This histochemical stain provides a simple and accurate means of determining the telogen/anagen ratio in plucked hairs.


Allergic contact dermatitis to the perfume in Mycolog cream. Mycolog contact allergy is due primarily to its ethylenediamine and fragrance components. In the evaluation of Mycolog-sensitive patients, the major constituents, including the perfume at 5% or 10% in petrolatum, should be tested.


Pinguecula and actinic elastosis. An ultrastructural study. Three cases of pinguecula (conjunctival elastosis) were ultrastructurally investigated. Findings were compared with the features described in cutaneous actinic elastosis. Changes observed were not uniform. In both diseases, numerous elastotoc fibers were present with a finely granular matrix and masses of dense grains. The degenerative changes of the elastotic fibers evolved differently in conjunctival stroma and in dermis. The superficial extracellular concretions observed in pinguecula seemed to be an ultimate stage of elastotic degeneration. Like actinic elastosis and elastosis observed in chronic radiodermatitis, pinguecula is believed to result from a dystrophic increased elastogenesis induced by chronic irradiation, with secondary degenerative changes.


An exploration of a patient-centred nursing approach in the admission of selected surgical patients: a replicated study. It is well recognized that admission to hospital is a stressful experience for patients, and little attention is directed towards helping patients to adapt to hospitalization. This paper reports the replication of a descriptive study which explored the patient's welfare after the use of a patient-centred nursing approach designed to alleviate stress on admission to hospital. It also attempts to determine the patient's perception of the nurse during the implementation of the admission procedure, and his general reactions toward the admission procedure.


Learning disabilities: the role of the developmental optometrist. The role of the optometrist in treating children experiencing learning disabilities embraces numerous areas beyond the customary optometric services. Etiological, diagnostic and therapeutic factors are discussed stressing visual functional disorders, perceptual-motor and developmental lags and cognitive style. Their effects on the learning disabled child are analyzed; and the rationale of optometric procedures frequently used in treating LD children is reviewed. The optometrist may also be helpful in counseling patients who have learning readiness problems, reading disorders, psychological difficulties and nutritional deficits.