Immunoperoxidase localization of alpha bungarotoxin: a new approach to myasthenia gravis. The IPBT method has made it possible to precisely visualize the AChR. Normal distribution of AChR is at the peaks of the postjunctional folds of the muscle sarcolemmal membrane with a small amount present on the axonal tip as well. Denervated muscle fibers have extrajunctional AChR. In MG, there are also denervated-appearing fibers but these do not have extrajuctional AChR with the IPBT stain. To explain this, we have been able to demonstrate a serum factor capable of blocking the binding of alpha-BuTx to the AChR and have shown for the first time that this factor is capable of acting at the neuromuscular junction itself. This blocking factor may play a major role in causing the weakness of MG.


Morphological and histochemical changes of motor units in myasthenia. Neuromuscular biopsies were obtained from 45 myasthenic patients. Motor innervation was studied in all specimens by vital staining with methylene blue. Quantitative data included the proportion of elongated motor endings, and the terminal innervation ratio (TIR) of motor axons. Quantitative histochemical data, obtained on 12 biopsies, included the atrophy factors of type I and II fibers, the I/II ratio, and the proportion of fibers strongly reacting to both ATPase and NADH diaphorase (type III fibers). Statistical analysis of the data led to the following conclusions: (1) elongated motor endings, found in  26 biopsies, were not related to denervation or to the severity of the disease, and were preferentially observed in younger patients; (2) increased TIR suggesting denervation was observed in 7 biopsies, only in patients over 50 years; and (3) various histochemical changes were found, mainly numeric reduction of type II fibers, having no demonstrable relationship with the incidence of elongated motor endings. The highest TIR was observed in a biopsy containing an increased proportion of type III and intermediate muscle fibers.


Polyfunctional antibodies. Their biology and inheritance. It seems likely that immunoglobulins have evolved from some archetypal molecule and those forms which are useful to the animal have been retained. It is this entire population of antibodies which forms the humoral immune system and in such a system, not only the properties of individual antibody combining regions, but the properties of the multiprotein system as a whole, are important for the defences of the body against pathogens. Antibody combining sites may bind a disparate set of structurally related and unrelated ligands. This multispecificity can be biologically meaningful: the same clone can be stimulated by different antigens. In this sense, cell surface immunoglobulins are multifunctional. The major biological consequence of antibody multispecificity is overlapping binding functions within subsets of the total antibody repertoire. The most significant impact of this overlap is: (1) it reduces the number of V genes necessary to code for the total number of combining sites; (2) the cross-stimulation of clones by structurally related and unrelated antigens may be instrumental in the normal maintenance of immune responsiveness and in addition, it may explain the ability to respond to unusual and less ubiquitous antigens; (3) the antigenic history of the animal may contribute to the maturation of the immune response by cross-stimulation of pre-selected clones of antigen binding cells.


The amino acid sequence of the variable regions of the light chains from two idiotypically cross reactive IgM anti-gamma globulins. Amino acid sequence analysis of the light chains of two IgM rheumatoid factors is presented. The parent molecules of these chains are human IgM proteins with specificity for human IgG and sharing idiotypic cross reactivity. The heavy chains had previously been shown to have extremely similar hypervariable regions. This communication presents data indicating that the light chains, even though representing two different kappa subgroups, also share identities in their hypervariable regions. These data can be most easily interpreted in terms of a gene interaction model for immunoglobulin V region assembly.


The relationship between hypervariable regions, antigen-binding specificity and the three-dimensional structure of antibodies. The amino acid sequences of the V domains (VL + VH regions) of 3 antibodies raised to type III pneumococcal polysaccharide in individual outbred rabbits are reported. With the exception of the second hypervariable section of the L chains, these antibodies have very different sequences in the hypervariable segments of the V domains. Within the third hypervariable region of the H chain, each antibody has a different length. On the sole basis of the amino acid sequences of these three anti-pneumococcal antibodies, the results do not support the concept of a simple correlation between primary structure in the hypervariable sections (known to determine the shape of the combining site) and antigen-binding specificity. In view of the large number of amino acid interchanges in the hypervariable positions and because of the size difference between the rabbit H chains, it is likely that each anti-SIII H chain studied here represents the product of a different structural germ line gene. In contrast to such a complex immune response to a relatively simple polysaccharide antigen, an extensive structural uniformity has been observed in the heavy chains of mouse myeloma proteins with phosphorylcholine activity and a mu chain from a human Waldenström IgM endowed with the same activity. The finding of a very similar heavy chain variable region in two different species which are separated by about 75 million years in evolution favours the concept of stable transmission of variable region genes throughout evolution.


Structural and functional mapping of immunoglobulin V-regions. V-regions of immunoglobulins chains contain 3 types of positions, which are equally represented: invariant and sub-group characteristic, which account for the "framework" and hypervariable positions, responsible for antigen recognition. The 3 types of positions are grouped and fall within a very few discrete stretches. Sub-group characteristic segments containing one of the 2 cysteyl residues of the V-regions may be isolated by high voltage paper electrophoresis and provide a basis to type for sub-groups in the VK and in the VH human systems. This allowed to characterize a large set of human myeloma proteins that were used in a series of competitive hybridizations which indicated that sub-groups had no influence on preferential reassociations, which occurred in 80% of the cases. This preference seems to rely mostly on individual structural differences, which may be linked to heterogeneity at the framework level. Distinction between framework heterogeneity and hypervariable regions heterogeneity may be approached by raising antibodies against a mouse myeloma protein, MOPC 173, of known sequence, by means of syngeneic and allogeneic immunizations, using Balb/c and A/J mice. Junction of distinct portions of immunoglobulin chains such as the V and the C regions raises the possibility that some recognition signals may operate at the DNA level. Since rotational symmetry regions in the DNA are known to act as such signals, it is discussed whether such regions can be expected from the amino acid sequence data, especially in the vicinity of the "switch" peptide.


Heavy and light chain variable region subgroups: antigenic analysis among human monoclonal immunoglobulins. An antigenic analysis of human heavy and light chain variable region subgroups has been possible by means of heterologous specific antisera using a hemagglutination-inhibition procedure. The specificity of antisera had been shown to be directed against antigenic determinants of VkappaI, VkappaII, VkappaIII, VlambdaI, VlambdaII, VlambdaIII and VHIII by means of chemically subgrouped proteins. A series of IgG, IgA, IgM and IgD were examined for the presence of a VHIII variable region subgroup antigenic determinant. The data showed that 50% IgG, 62% IgA, 55% IgM, 45% IgD were positive for VHIII antigenic determinant. NH2 terminus blocked monoclonal immunoglobulin belonging to the VHIII subgroup were found, increasing incidence of this subgroup among IgG. A preferential association of VHIII antigenic determinant with IgG1, IgG3 subclasses was observed among IgG myeloma proteins while the preferential association was only observed with IgG1 subclass when anti-Rh antibodies were considered. Among 53 IgG of kappa type 20 (37%) kappaI, 30 (56%) kappaII, 3 (5%) kappaIII subgroups were found and among 42 IgG of lambda type 13 (31%) lambdaI, 13 (31%) lambdaII and 16 (38%) lambdaIII were observed. Although numbers are limited in each group for conclusions a study of VHIII and non VHIII antigenic subgroup determinant with respect to the light chain subgroups is given. The non-allelic behaviour of this VHIII antigenic determinant was observed. Preliminary data on the presence of these human antigenic determinants among different animals species were given.


Phylogenic studies on allotypic specificities of rabbit immunoglobulins. Phylogenic studies of rabbit allotypic specificities are carried out in several lagomorphs. We give the results obtained through the study of hare Lepus capensis immunoglobulins. IgG of all the 140 studied hares exhibit determinants recognized by anti-b4 and anti-b5 immune sera. Most hares, but not all, have determinants recognized by anti-b6 immune sera. Among the determinants recognized by anti-b4 immune sera, some seem to be isotypic (found in all studied hares), others (absent in some hares) are allotypic. Determinants recognized by anti-b5 immune sera behave as isotypic; they are recognized by anti-b5 antibodies which participate to the cross reaction with b6 pattern. Other determinants recognized by anti-b5 antibodies are allotypic. Three allotypic patterns, at least, each made of two determinants, belonging to the same series, have been detected on hare L chains. Two of them are recognized by anti-b6 immune sera; the third is characterized by its lack of determinants cross-reacting with anti-b6 antibodies. It does not seem that L. capensis has any determinant recognized by anti-b9 immune sera. IgG of some but not all hares have determinants recognized by anti-a2 immune sera, only when these have been prepared in rabbits with the phenotype a(1-2-3+).


Allotypes of the a series and their variants in rabbit immunoglobulins. Six allotypic specificities of the a series are found on rabbit immunoglobulins: a1, a2 and a3 are found both in domestic and wild rabbits Oryctolagus cuniculus; a100, a101 and a102 seem to be present only in wild rabbits. Each of these specificities is a family of variants always present together in a given serum. These variants can be studied through the cross-reactivities detected between the patterns of the a series. The results of studies of cross-reactivities between a1, a3 and the two specificities a100, and a102 and also the cross-reactivity between a2 and a minor variant of the a1 specificity suggested a hypothetical scheme. This hypothesis attempts to take into account the evolution of the specificities of the a series and their variants. This hypothesis also postulates the existence of a set of closely linked genes which control the synthesis of the variants of a given specificity. One could suppose that primordial allelic genes might have appeared from an ancestor gene. By duplication each allele would have led to the appearance of a set of genes coding for a given specificity. These genes might have evolved through mutations and recombinations. In wild rabbits, the observation of an allotype which seems to result from a recombination between the group of genes coding for the a2 variants and the group of genes coding for a3 variants argues in favors of the genetic recombination mechanism.


The V gene repertoire as revealed by polyclonal B cell activators. The evidence for the one non-specific signal hypothesis stating that the B lymphocytes are activated by non-clonally distributed receptors which are not the Ig receptors, have been summarized. Even though protein A is a polyclonal B cell activator, it does not exert its effect by interacting with the Fc part of Ig receptors. One consequence of the one non-specific signal concept is that thymus-dependent antigens cannot activate or tolerize B cells. It was shown that B cells from animals tolerant to a thymus-dependent protein antigen could be activated by polyclonal B cell activators to produce antibodies against the tolerogen. Experimentally induced tolerance did not differ from tolerance to self antigens, since LPS and PPD induced autoantibodies capable of lysing autologous red cells and isotope labelled autologous and syngeneic spleen cells. Thus, B cells cannot discriminate self from nonself whereas T cells have been shown to possess this ability.


Assessing B cell diversification by antigen receptor and precursor cell analysis. A major element in the understanding of B cell specificity diversification is the extent of diversity present in mature and in developing B cell populations. Two general methods are currently used for assessing the specificity repertoire: (a) the enumeration of cells whose receptors can bind a specific antigen, and (b) the enumeration of cells which can respond to antigenic stimulation by antibody-forming cell clone production. Our laboratory has utilized the latter method to establish the frequency of B cells responsive to a wide variety of antigenic determinants. The findings indicate that: (a) the primary murine B cell specificity repertoire probably includes more than 10(7) clonotypes; (b) some clonotypes are represented by numerous B cells (40,000 TEPC 15 precursors per BALB/c mouse) while most are represented by fewer than to B cells per mouse; (c) the acquisition of the repertoire is apparently antigen-independent since germfree mice have repertoires similar to conventional mice and secondary B cells are easily distinguished from primary B cells; (d) the neonatal repertoire appears to contain only 10(4) clonotypes at birth, each represented by perhaps 200-400 cells; (e) the diversification process from neonatal to adult repertorie appears highly ordered and reproducible. Antigen binding cell studies have now used in conjunction with the splenic focus assay in an attempt to correlate these two techniques. The results indicate that the efficiency of the splenic focus assay used for precursor cell anlysis it 4-5% for both primary and secondary B cells and is similar to the percent of donor B cells lodged in recipient spleens. For certain antigens (DNP-BSA) the number of antigen-binding cells can represent 4% of the total B cells, and this number directly correlates with the concentration of antigen used; stimulation, on the other hand, appears to have an affinity threshold achieved by only 0,02% of the DNP-specific B cells. In contrast, PC-BSA antigen-binding cell and splenic focus precursor cell frequencies are identical. These findings are interpreted to indicate that antigen-binding cell analyses confirm the validity of the calculations used to estimate precursor frequencies in the splenic focus technique. However, for some antigens binding to cell receptors, one detects a large number of cells belonging either to a nonstimulatable B cell subclass or whose receptor affinity is too low to permit stimulation.


Inhibition of specific binding of DNP (dinitrophenyl) determinant to lymphoid-cell receptors by related and unrelated compounds : quantitative studies in vitro. The effect of related and unrelated compounds on the specific binding of dinitrophenyl-coupled bacteriophage (DNP-T4) to lymphoid cell receptors has been examined and compared with the effect on the neutralization of DNP-T4 by anti-DNP serum. Spleen cells and sera from Balb/c mice immunized with DNP-bovine serum albumin were used. The binding of DNP-T4 to the cells was inhibited by DNP-eAcp, di-DNP-Lys, DNP-Tyr, DNP-p(Ornith) and DNP-BSA (among the DNP-derivatives tested), TNP-BSA, ARS-p(Tyr) and TGA. In addition with the above named DNP and TNP compounds, the DNP-T4neutralization by antiserum was also prevented by DNP-derivatives with either L-cysteic acid, alanine, glutamine or poly-L-glutamic acid, while ARS-p(Tyr) and TGA were not effective. Plain carriers (BSA, HSA, poly-ornithine, polylysine and polyglutaminc acid) and cell-mitogens (ConA, LPS and PPD) had no significant inhibitory effect. The results obtained indicate the occurrence of differences between cell-bound receptors and circulating antibodies in what concerning their specific reaction with the dinitrophenyl determinant.


Development of immuneeoglobulin and antibody-forming cells in different stages of the immun response. After primary immunization of mice, rats and rabbits with antigens (horse radish peroxidase, bovine serum albumin and muchroom tyrosinase) emulsified in complete or incomplete Freund's adjuvant, both cells synthesizing immunoglobulin without detectable antibody function and antibody-producing cells were detected. The first cells which appeared were synthesizing and secreting IgG and IgM immunoglobulins without antibody function. These cells were progressively replaced by cells synthesizing and secreting antibodies. In some plasma cells of mice, rats and rabbits immunized with peroxidase, antibody activity was detected only in restricted areas of the cytoplasm ; the remainder contained antigenic determinants of immunoglobulins. After secondary immunization the results were the following: in mice, both cells containing immunoglobulins without antibody function and antibody-containing cells appeared simultaneously and they were present in equal amount; in rats, only the antibody-containing cells were present in high number. Immunizations performed using different protein antigens (horse radish peroxidase, human and bovine serum albumin, aggregated and desaggregated human IgG, and ovalbumin) injected as a solution in saline have shown that after antigenic stimulation both populations of cells appeared, their number depending on the dose of the antigen injected. Further experiments carried out with tolerant mice, with germ-free animals and with "B" mice have shown that the appearance or not of antibody-producing cells was always related with respectively the presence or absence of cells synthesizing immunoglobulins without detectable antibody function. Finally experiments performed on rabbits have shown that some cells containing immunoglobulins without antibody function share idiotypic determinants in common with cells synthesizing antibodies.


The biological origin of antibody diversity. Antibody diversity has a compelling fascination for many scientists and over the years speculations have sometimes seemed more numerous than facts. Now the structural basis of antibody specificity is well defined. Amino acid sequences and recently three-dimensional structures of various immunoglobulins provide the most solid basis for discussing the origin of diversity. The novel pattern of variable (V) and Constant (C) regions of amino acid sequence has been resolved further to show the functional pattern of variability. Inheritance of separate V and C genes is accepted, but attempts to define more than one gene coding for each V region are considered here to be unnecessary. The pattern of variability is still best understood in terms of mutation and the presence or absence of various selective pressures. The major area of debate still hinges around the extent to which mutation and selection operate during evolution or somatically. Sequence data have now been generally interpreted to require multiple V genes carried in the germ line. A few individual VH genes have been mapped in close linkage to CH genes in the mouse. The apparent existence of three VH alleles in rabbits was a strong argument against multiple V genes. Now the three phenotypes have been shown to be due to alleles controlling the expression of three sets of VH genes all present on the same chromosome. That V-gene expression requires rejoining of V and C genes at the DNA level is now almost certain. Models for the joining process can draw on the precedents of transposable genetic elements, which are widespread in Nature. The total extent of antibody diversity remains a philosophical point. Estimates of the number of antibody molecules required for observed diversity are reduced by two recently documented proposals. Each antibody combining site apparently has many (estimated at 100) different specificities and most combinations of VH and VL regions probably form a viable site. A given combining site can be defined by its pattern of shared specificities. Several specific antibody repertoires have been measured and the size in each case is consistent with the stringency with which the specificity is selected. Repertoire size appears to be under genetic control, but there are problems in viewing the genotype through the veil of clonal selection. Molecular hybridization has been used recently in an attempt to count V and C genes directly. C genes are seen in DNA having nonreiterated sequences, as formal genetics predicts. Each V-region probe hybridizes at a similar rate to C-region probes. Interpretation of this result depends on the extent to which one V-region probe will reveal nonhomologous V genes. Previous estimates that many cross-hybridizing genes should have been seen if present are possibly exaggerated. It is argued here that the data are compatible with a germ-line gene for each probe studied. Maximum estimates for the number of germ-line genes are sufficient to account for antibody diversity...


Protease inhibitors in plasma of patients with chronic urticaria. The hypothesis that deficiencies of plasma protease inhibitors might play a role in the pathogenesis of chronic urticaria was evaluated. Plasma levels were measured in patients with urticaria and a matched control group for alpha1-antitrypsin, alpha2-macroglobulin, total trypsin-inhibiting capacity, kallikrein-inhibiting capacity, and the complement factors C1 esterase inhibitor, C3, and C4. A total of 92 patients with chronic urticaria or more than three months' duration was studied. Patients with acquired cold urticaria had significantly decreased levels of alpha1-antitrypsin and total antitrypsin activity. In patients with acquired angioneurotic edema, alpha1-antitrypsin levels and antichymotrypsin activities were lowered, with less significant decreases in anti-trypsin and antikallikrein activities. Levels of C1 esterase inhibitor , C3, and C4 were normal in all groups. There was no correlation between the increased sensitivity to intracutaneously administered kallikrein injection and deficiencies of of protease inhibitors.


Chemosurgery for skin cancer: fixed tissue and fresh tissue techniques. Complete microscopical control of the excision of cancer of the skin is achieved by removing tissues layer by layer and examining the undersurface of each layer by means of frozen sections. If the cancer is extensive and complicated or if it is of a type readily spread over an excisional surface, the tissues are fixed in situ with zinc chloride prior to excision (chemosurgery, fixed tissue technique). If the cancer is not too extensive or complicated, the tissues are excised in the fresh, unfixed state (chemosurgery, fresh tissue technique). Both techniques are highly reliable; for example, in two consecutive series of basal cell carcinomas treated chemosurgically, the five-year rate of cure was 99.3% for the 9,351 lesions removed by the fixed tissue technique and 97% for the 127 lesions removed by the fresh tissue technique.


Granulocyte transfusion in treatment of infected neutropenic children. Thirty episodes of infection in severely neutropenic children with acute leukaemia and aplastic anaemia resistant to antibiotics were treated with total of 51 granulocyte transfusions collected from normal donors by means of an Aminco continuous flow cell separator. The mean granulocyte increment in the recipients was 820/mm3 at 1 hour and 307/mm3 at 15 hours post-transfusion and it showed a correlation with the number of cells transfused, the size of the child, and his pretransfusion granulocyte count. Fifteen (50%) episodes responded clinically by 24 hours and 23 (77%) by 5 days post-transfusion. Two more children recovered more slowly and 5 died. Poor response was associated with multiple-site infections and with a prolonged period of infection before granulocyte transfusion, but the primary diagnosis did not influence the outcome. We conclude that granulocyte transfusion significantly reduces the mortality and morbidity from infection in neutropenic children with acute leukaemia.


Lipoprotein lipase in the fat body of Triatoma brasiliensis. Lipoprotein lipase activity was detected and characterized in the fat body of  Triatoma brasiliensis one of the vector of Chagas' disease. The enzyme activity was close to a free fatty acid production of 47 mumol FFA mg protein-1-h-1 when assayed in the complete system. 2. The enzyme presented maximum activity at pH 8.5. It appeared to be activated by heparin (2 U/ml) and to require a cofactor (rat serum lipoproteins) when assayed with simple triglyceride emulsions. The requirement for a FFA acceptor (albumin or CaCl2) was also observed. 3. Inhibition of enzyme activity by pyrophosphate, HDB, protamine sulphate and 1 m NaC1 indicated further that the enzyme from fat body of T. brasiliensis had the characteristic features of lipoprotein lipase from other sources.


[Effect of cortisone acetate on acid secretion induced by histamine, pentagastrin, gastrin and food in dogs]. The effects of long term treatment with cortisone on the gastric secretion induced by histamine, pentagastrin, porcine gastrin and a meal have been investigated in four dogs with both gastric fistula and Heidenhain pouch. Cortisone increased the postcibal acid output and the observed maximal acid response from the  pouch to all three exogenous stimuli. The ED 50'S  remained unchanged. The same effects although less marked were observed in the innervated stomach. These data indicate that the increased acid secretion observed after long term treatment with cortisone is largely due to an increased secretory capacity of the gastric mucosa. This latter could result from an increase in the number of secretory units or to partial removal of a non competitive inhibitor of gastric secretion.


Changes in thoracic and right duct lymph flow  and enzyme content during skeletal muscle stimulation. In the pentobarbital-anaesthetized dog the effect of electrical stimulation of hindlimb skeletal muscles on thoracic and right duct lymph flow and  enzyme content was examined. Increase in plasma creatine kinase (CK), L-aspartate-aminotransferase (AST) and lactic dehydrogenase (LDH) during 30-min muscle stimulation were not significantly altered by draining lymph. Both right duct and thoracic duct lymph flow trebled during stimulation. At the same time, the activity of the three enzymes examined decreased in right duct lymph and increased in thoracic duct lymph. Of the latter,  only the increase in lymph CK was of a sufficient magnitude to have resulted in a detectable increase in plasma CK. CK was the smallest of the three enzymes studied and apparently preferentially entered the lymph, suggesting that the larger AST and  LDH molecules were not likely to have entered the blood plasma directly from skeletal muscle. Rather their entry from some other  tissue, possibly the formed elements of the blood, is indicated.


Effect of certain organic  compounds on saliva secretion in sheep. In two sheep fitted with esophageal cannulas the stimulation of resting saliva secretion by intrabuccal injections of volatile fatty acids (VFA) and of certain organic plant compounds was investigated. Volatile fatty acids, salicin and vanillin increased saliva flow rate by 100, 76 and 72% over 2.5 min after injection. Minor positive or negative effects were obtained with quercitrin, protocatechuic acid, grass juice, anethole and agaricinic acid. Injections of volatile fatty acids and of salicin induced chewing activities, which were significantly correlated with salivary flow rates. The results indicate that organic plant compounds can provide a stimulus for saliva secretion mediated in part by chewing activities.


Study of messenger-like RNA extracted from HeLa cells polysomes. II. Identification of the types of RNA eluted from  methylated albumin kieselguhr columns. Together with the elution pattern of pure messenger RNA molecules of various origin, the labelling kinetics of rapidly labelled heterogeneously sedimenting RNA (HSRNA) extracted from polysomes of HeLa cells have been studied by chromatogrphy on columns made of methylated bovine serum albumin adsorbed on kieselguhr. HSRNA is eluted within three peaks-IP, Q2P and TDP-following in that order the increase of NaC1 concentration in  the eluting buffer. Besides peak TDP which results from an experimental artefact, our data suggest that the appearance of peaks IP and Q2P reflects the absence and presence respectively of polyadenylic acid stretches in these molecules. Within peak Q2P, the critical factor affecting the order of elution  is the size of the polyadenylic acid stretch.


Characteristics of hypotension elicited by electrical stimulation of the lateralhypothalamusin anaesthetized dogs. Electrical stimulation of the lateral hypothalamus near the paraventricular nucleus hypothalami, resulted in : hypotension, light bradycardia, decrease of the left ventricular systolic pressure without increase of the left ventricular  end-diastolic pressure or left ventricular output, femoral vasodilatation and occasionally renal vasodilatation. The hypotensive reactions were potentiated by baroreceptor deafferentation. They were not blocked by anticholinergic, antihistaminic or antidopaminergic agents.


Hypothalamic depressor reactions in anaesthetized dogs after baroreceptor exclusion and during norepinephrine infusions. The stimulation of the paraventricular hypothalamic centre inhibits the vasoconstriction of the peripheric arterioles and the myocardial effects elicited by endogenous or exogenous increased catecholaminaemia.


Susceptibility to insecticides of Lasioderma serricorne (F.) (Coleoptera anobiidae) grown on vitamin-deficient diets. Variations in the susceptibility to pyrethrum and chlordane (two contact insecticides) of Lasioderma serricorne grown on different vitamin-deficient diets were studied and reasons for such variation were proposed.


Acute chlordane toxicity on the serum alkaline phosphatase activity of Meriones hurrianae Jerdon. Different acute doses of chlordane enhance the serum alkaline phosphatase activity in Indian desert gerbils. The damage to parenchymal cells of liver, and hepatic microsomal enzyme induction as a result of chlordane treatment are discussed as the possible reasons for the increase in serum alkaline phosphatase activity.


[Effect of curarization on sensitivity of chicken latissimus dorsi to electric current]. (1.) To test the sensitivity to electric current of the latissimus dorsi and, therefore, its ability to yield action potential, we have studied the effect of curarisation on the amplitude of the isometric tetanus during "massive" stimulations by alternative current. (2.) Curarisation causes a considerable but reversible  reduction of the tetanic tension of the latissimus dorsi anterior (L.D.A.) but has no effect on the tetanus of the latissimus dorsi posterior (L.D.P.). It is concluded that a part of the L.D.A. is not sensitive to electric current and is thus  unable to yield action potentials, unlike the opinion of some authors. (3.) Rising the temperature has an inverse effect on the speed of the tetanus ascending phase, depending on whether the L.D.A. has been curarised or not. This show the existence of two types of excitation processes of very different nature in this muscle. (4.) Direct current is much less efficient than alternative current on the curarised L.D.A.


Chronotropic responses to experimental ischemia of the canine sino auricular node. (1) Using isolated, blood-perfused atrium preparation of dogs, the effect of ischemia on sinus rate was studied in ten preparations. Cessation of atrial perfusion usually induced gradual deceleration of the sinus rate which was not blocked by atropine. Occasionally, brief and slight sinus acceleration was initially observed in three of ten atrium preparations. This positive  chronotropic effect was not  blocked by a  beta-adrenoceptor blocking agent, propranolol. (2) In every preparation, just after release of occlusion, there was an initial profound sinus deceleration, occasionally followed by oscillatory changes in sinus rate. (3) The chronotropic response pattern induced by temporary occlusion and release of the sinus node artery was not influenced by propranolol, phenoxybenzamine or atropine treatment. (4) These results suggest that ischemia exerts its principal effect directly on the sino auricular node pacemaker cells, rather than on neighboring nerve endings.


Variations of rat brown adipose tissue composition during cold acclimatization. The modifications in weight and composition (lipids, proteins, water) of rat interscapular brown adipose tissue (BAT) were studied along the six first weeks of cold exposure and acclimatization. The variations of noreponephrine content was also investigated. During the first day of cold exposure, the major part of tissue lipids was released. During the following two days there was a fall in lipid and norepinehprine contents and uptake of water. Then, until the end of the first week a  rapid repletion occurred. At that moment the relative pass of the tissue and the amounts of its principal components reached values which are not changed during the following weeks. We can conclude that the adaptative changes in the levels of BAT essential components are carried out at the end of the first week of cold exposure, long time before the non shivering thermogenesis is entirely effective.


Post-tetanic changes of depressor responses evoked by stimulation of the aortic nerve. The effect of changes in duration of the conditioning tetanus on the size of the testing depressor  response was studied in rabbits anaesthetized with urethane. Depressor responses were evoked by stimulation of the aortic nerve. The interval between the conditioning and testing stimulation was fixed at 40 and 120 sec. Two frequencies of conditioning tetanus were employed. Brief conditioning tetani facilitate the testing response. With lengthening of the conditioning stimulation the size of the testing response is decreased and when duration of the conditioning amounts to 20-60 sec depression of the testing fall of blood pressure reaches a steady level. Further increase in duration of the conditioning tetanus to 180 sec does not affect the plateau of depression of the testing response. The longest duration of conditioning affecting the size of the testing response is  considered to determine the range of control of the testing response exerted by preceding conditioning tetanus. Since the levels of plateau of depression are different for two used frequencies of conditioning, it is suggested that this factor may control the size of the testing response beyond the range of control executed by the duration of the conditioning tetanus.


[Plasma concentration curves of catecholamines and corticosteroids as well as PBI in swine kept under various conditions and acutely exposed to cold]. The response to acute cold (-5 degrees C) of swine kept single was found to differ from that of animals kept in groups. Such difference in reaction was reflected in significant variation of plasma concentration curves in catechol amines and 11-OHKS during and after acute exposure to cold. The response to cold stimulation of animals kept single was primarily and pronouncedly via the sympathicoadrenal system, whereas animals kept in groups responded to acute cold exposure, with the hypothalamic-adrenocortico-pituitary system playing the major role. Significant rise in PBJ in response to 30 and 90 minutes of exposure to cold was recorded only from animals kept single and interpreted as an expression of strong adrenergic reaction of such probands. Such difference in response to one and the same acute stimulus supported the conclusion that there was a difference in adaptation capacity between animals in those two keeping conditions. The results also suggested that type and degree of organic response to a specific stimulus were likely to depend not only on the intensity of the given stimulus, but also on the control potential produced by all closely interconnected control circuits at the time of stimulation.


Antigenic characterization of c-type oncornavirus isolated from lymphomatous baboon (Papio hamadryas). Antigens of BILN C-type oncornavirus isolated from lymphomatous baboon were studied in the double immunodiffusion. The major viral protein (P-30) of this virus possesses interspecies antigenic determinants (GS-3). This protein also revealed close antigenic relation to the P-30 of RD-114 and M-7 C-type oncornaviruses. Specific serum against BILN P-30 has been produced. The comparison of the three test-system (P-30s of M-7, RD-114, and BILN, and specific sera against them) did not allow to distinguish these proteins by the method used.


[Impulse cytophotometric investigations on tumour material (author's transl)]. By impulse cytophotometry (ICP) we can get the distribution of the frequency of cell nuclei as a function of the fluorescence intensity. The interpretation of the curves is only possible on the basis of histophysical and histochemical features. The following problems are discussed: specificity of staining with ethidium bromide, pretreatment, detritus, comparison with other independent methods. The second peak of the karyogram consists of G2-cells and, in polyploidy of tetraploid cells. This peak is often high in cell suspensions of malignant tumours; sometimes we found polyploidy up to 16c. The stem line concept is discussed. The ICP is a valuable aid in exfoliative cytology, but no screening method.


[Morphological studies on cellfree induced sarcomas in syrian hamster (author's transl)]. Morphological studies on sarcomas induced in syrian hamsters by cellfree transmission are described. The tumour tissue for the cellfree preparations stemmed from a sarcoma, containing C-particles. Basically, three histological groups have been distinguished: 1. neoplasms of the peripheral nerve-sheath, 2. undifferentiated sarcomas, and 3. liposarcomas. Furthermore, a rhabdomyosarcoma, an angiosarcoma and, in a heterotransfection on rat, an osteosarcoma have been established. The great majority of tumours could be transmitted by cellfree preparations. To this neoplasms belong the undifferentiated histological structure.


Formed blood elements in peritoneal effusion and the macrophage migration inhibition test in healthy guinea pigs and in guinea pigs with experimental allergic encephalomyelitis. An attempt was made to correlate the percentages of macrophages, lymphocytes and granulocytes in the peritoneal effusion in healthy guinea pigs and guinea pigs with experimental allergic encephalomyelitis (EAE), with the macrophage migration inhibition (MMI) test. Varying percentages of the cells had no influence on values of MMI. Similarly, in guinea pigs with EAE, percentages of formed elements in peritoneal effusion were not correlated with intensity of MMI or with histopathologic lesions in the brain and spinal cord. It is suggested that the observed differences are due to individual immunologic responsiveness of animals and, probably, to other hitherto unknown mechanisms.


Influence of the alpha-globulin fraction isolated from calf serum on graft-versus-host reaction in strongly histoincompatible mice. The influence of a two alpha-globulin fractions isolated from calf serum by the method of Mowbray on the graft-v-host reaction (GVH) in the F1 hybrids of inbred mouse strains differing  in H-2 locus after administration of parental lymphatic cells was studied. Some of the mice were X-irradiated with 500 R before administration of parental cells. The studied fractions exerted strong suppressive action on the splenomegalic reaction and proliferative phenomena in the spleen, accompanied by intensification of changes in the liver, particularly in irradiated mice, It was concluded that the applied doses of the investigated preparations had no influence on immunologic actvity of the donor's lymph cells, but that they inhibited nonimmunologic proliferation of recipient cells and consequently impaired defense mechanisms in the F1 hybrids.


Further studies on the influence of calf alpha-globulin on survival of skin allografts in mice. A single injection of 10 mg of alpha-globulin fraction isolated from calf serum prolonged survival of skin allografts in mice provided the preparation was injected two days before grafting. An immunosuppressive effect was also obtained by injecting alpha-globulin in a total dose of 13 mg in the second week after grafting in mice pretreated with antithymocyte serum; On the other hand, such an effect was not obtained when alpha-globulin was injected in the early days after grafting in doses of up to 25 mg per mouse.


Ribonuclease treatment of tick-borne encephalitis. Clinical trials of the pancreatic ribonuclease (RNAse) in treatment of patients with tick-borne encephalitis were carried out. Two hundred forty-six patients were treated with RNAse and 261 were given antiencephalitic gamma globulin. Administration of 30 mg of RNAse every four hours over a period of five to six days provided stable high concentrations of the enzyme in blood that exceeded normal values by two to three times. An increase of RNAse activity in cerebrospinal fluid also took place. The comparison of RNAse and gamma globulin demonstrated a considerable advantage of the former: under RNAse treatment the temperature normalized on the average of twice as quickly, and a rapid regression occurred in meningeal symptoms and cerebrospinal fluid pleocytosis. In patients treated with RNAse, no adverse reactions or aftereffects were observed. On the grounds of the reported results, RNAse was accepted officially as a new, highly efficient antiviral drug for the treatment of tick-borne encephalitis.


The chemical composition of wool. XV. The cell membrane couplex. The cell membrane complex of wool has been examined by electron microscopy of stained cross sections after immersion of the wool in formic acid. The cell membrane complex of the cortex is considerably modified by the treatment, but that of the cuticle appears unchanged. Resistant membranes from cuticle cells, cortical cells and wool have been prepared by treatment with performic acid-ammonia. Amino acid analyses show that the resistant membranes from the cuticle contain citrulline but those from cortical cells do not. It is concluded that the cell membrane complex of the cuticle differs from that of the cortex. Because of the high lysine content of the resistant membranes, their resistance to chemical attack, the hydrophobicity of epicuticle and the observation of a small amount of epsilon-(gamma-glutamyl)lysine, it is postulated that the resistant membranes may contain an appreciable amount of epsilon-(gamma-glutamyl)lysine cross links.


Occurrence of alpha-1-fetoprotein in the mammary gland tissue of men with gynecomastia. Immunohistochemical proof of A1FP production in the mammary gland of a man with gynecomastia is published here for the first time. The immunohistochemical proof of this fetal alpha-1-globulin in the intraductal epithelium of the mammary gland is described on the basis of the case history of a 25-year-old man with a metastasizing malignant teratoblastoma of the gonads. The question, to what extent A1FP-neosynthesis is characteristic for malignant tumors or whether it is a cellular response to specific, in this case hormonal growth stiumli of primary tumors, is also discussed.


A smear technique for staining elementary bodies of fowlpox. Red-stained elementary bodies of fowlpox virus were detected in smears from lesions of infected chorioallantoic membrane and from skin of infected birds stained by the Giménez method.


Endocrine and metabolic indices during progestin (quingestanol) therapy. Quingestanol at 300 y/day was well tolerated during 1 to 12 months of therapy of pre-menopausal and menopausal women with minimal changes, if any, in  a battery of routine endocrine and metabolic indices. Serum inorganic phosphorus levels were slightly above the pre-therapy range but still within normal limits during the 12 months of treatment and the mean relative blood volume or hematocrit during the latter 6 months was slightly above the starting value.


The relation between electrophoretic mobility and serological subtype of HBSAg. The electrophoretic mobility of HBsAg in samples of serum has been correlated with subtype of the antigen. HBsAg samples have been placed in three major groups (ad, ay and 'intermediate') by both methods, and there has been 81.6% agreement between subtype and electrophoretic mobility.


Absence of binding of pancreatic and urinary kallikreins to alpha 2-macroglobulin. Pancreatic and urinary kallikreins failed to form the typical serine proteinase complex with alpha2M (alpha2-macroglobulin). Studies were performed to compare this with the binding of trypsin to alpha2M at various molar binding ratios, with the use of Sephadex G-200 gel filtration to separate free and alpha2M-bound enzyme fractions. The subunit conversion was totally absent with pancreatic kallikrein from lhich traces of a binding proteinase had been removed. The lack of binding is believed to be the result of the restricted specificity of the kallikreins.


[Effect of cromoglicinic acid on the degranulation of mast cells (author's transl)]. An interference microscopic method is described which allows quantitative measurements of the inhibiting effect of the di-sodium salt of 1,3-bis(3-carboxy-chroman-5-yl-oxy)-2-hydroxypropand (cromoglicinic acid; Intal; DSCG) on the degranulation of mast cells in vitro induced by polymixin B and Compound 48/80. The specific degranulation inhibiting effect of DSCG is increased in the presence of Ca++ and Mg++ ions.


'Incessant' tachycardias in Wolff-Parkinson-White syndrome. I: Initiation without antecedent extrasystoles or PR lengthening, with reference to reciprocation after shortening of cycle length. In 6 patients with the Wolff-Parkinson-White (WPW) syndrome, repetitive, almost continuous (incessant) reciprocating atrioventricular (AV) tachycardia has been shown to arise when the sinus cycle length was shortened to a critical point, at which unidirectional block occurred without the classical feature of PR prolongation. Though this phenomenon superficially resembles an aspect of chronic intranodal reciprocating tachycardia of children, basic differences can be identified. It was encountered more frequently in younger subjects; the only patient over 45 developed the arrhythmia as a complication of therapy. This incessant mechanism may explain some cases in which antiarrhythmic treatment does not control reciprocating tachycardia in the WPW syndrome, but such a mechanism can also occur spontaneously.


Tetralogy of Fallot. Risk factors associated with complete repair. Ninety-six consecutive total repairs of the tetralogy of Fallot are reviewed. There was an overall hospital mortality of 8-3 per cent and a total incidence of low output cardiac failure of 18-8 per cent, and this was the principal cause of death and the most important source of postoperative morbidity. In 49 cases there had been a previous palliative shunt procedure and there was a strikingly lower mortality and a highly significant lower morbidity in this group. Other factors which correlated in a positive fashion with increased mortality and morbidity were chronic hypoxia (as evidenced by polycythaemia), age below 5 years, severe postoperative right ventricular hypertension, and (to a lesser extent) extensive right ventricular outflow tract reconstruction. Data are presented to support the hypothesis that a palliative shunt procedure should be considered in the severely polycythaemic child with a surgically 'unfavourable' right ventricular outflow. This policy carries a low early mortality in our hands (5-4%), and is associated with a low mortality (3-9%) at a subsequent repair. This compares with a mortality of 12-8 per cent for primary repair, and the incidence of low output cardiac failure is five times as high in the primary repair as compared to the previously shunted group.


The detection by immunodiffusion of tumour associated antigenic components in extracts of human bronchongenic carcinoma. Antisera to extracts of a variety of bronchogenic carcinoma were raised in rabbits and extensively absorbed with immunoadsorbents prepared with normal lung extracts cyanogen bromide linked to Sepharose 4B, and glutaraldehyde insolubilized normal lung extracts. The antisera were tested by immunodiffusion against a panel of extracts from a variety of bronchogenic carcinoma, foetal lung extracts and pools of normal lung extracts. The results indicate that two distinct antigenic components are associated with bronchogenic carcinoma; one which is present in a high percentage of the tumour extracts tested and appears to have partial identity with a foetal lung component, and one (or more) which is not foetal and appears to have higher cross-reactivity (but not exclusively) with tumours of the same pathological type. Attempts to detect either antibody or antigens relating to these components in the serum of patients with bronchogenic carcinoma by these techniques were unsuccessful. The foetal cross-reacting component was neither carcinoembryonic antigen and alpha1-foetoprotein.


The effect of Corynebacterium parvum therapy on immunoglobulin class and IgG subclass levels in cancer patients. Detailed serological studies have been undertaken in a small group of cancer patients receiving nonspecific immunotherapy with Corynebacterium parvum (C. parvum). These patients included 4 cases of recurrent malignant melanoma, 2 of stomach cancer and 2 of recurrent breast cancer. They all received an initial i.v. infusion of 20 mg of a formol killed suspension of C. parvum followed by 2 mg (i.m.) at weekly intervals for 10-11 weeks. This protocol consistently resulted in an increase in the circulating IgG levels of all patients but had a variable effect on their IgA, IgM and IgE levels. Increases in the concentration of all 4 IgG subclasses contributed to the overall increase in IgG levels and these changes ranked IgG2 greater than IgG1 greater than IgG3 = IgG4. It also had an inconsistent effect upon the levels of alpha-macroglobulin in pregnancy but the levels of normal serum alpha2-macroglobulin were virtually unchanged. Pre-existing antibodies to C. parvum were noted in all the patients. Titres rose appreciably following C. parvum administration and remained at high, though fluctuating levels, throughout the 100-day period of observation. Absorption studies suggested that the development of antibodies to C. parvum accounted in part for the increased IgG levels noted following this form of therapy. The significance of these changes in relation to the possible anti-tumour effect of C. parvum is discussed.


Multi-temperature effects on Hill reaction activity of barley chloroplasts. 1. The relationship between temperature and Hill reaction activity has been investigated in chloroplasts isolated from barley (Hordeum vulgare L. cv. Abyssinian). 2. An Arrhenius plot of the photoreduction of 2,6-dichlorophenolindophenol (DCIP) showed no change in slope over the temperature range 2--38degreesC. The apparent Arrhenius activation energy (Ea) for the reaction was 48.1 kJ/mol. 3. In the presence of an uncoupler of photophosphorylation, methylamine, the Ea for DCIP photoreduction went through a series of changes as the temperature was increased. Changes were found at 9, 20, 29 and 36degreesC. The Ea was highest below 9degreesC at 63.7 kJ/mol. Between 9 and 20degreesC the Ea decreased to 40.4 kJ/mol and again to 20.2 kJ/mol between 20 and 29degreesC. Between 29 and 36degreesC there was no further increase in activity with increasing temperature. The temperature-induced changes at 9, 20 and 29degreesC were reversible. At temperatures above 36degreesC (2 min) a thermal and largely irreversible inactivation of the Hill reaction occurred. 4. Temperature-induced changes in Ea were also found when ferricyanide was substituted for DCIP or gramicidin D for methylamine. The addition of an uncoupler of photophosphorylation was not required to demonstrate temperature-induced changes in DCIP photoreduction following the exposure of the chloroplasts to a low concentration of cations. 5. The photoreduction of the lipophilic acceptor, oxidized 2, 3, 5, 6-tetramethyl-p-phenylenediamine, also showed changes in Ea in the absence of an uncoupler. 6. The temperature-induced changes in Hill activity at 9 and 29degreesC coincided with temperature-induced changes in the fluidity of chloroplast thylakoid membranes as detected by measurements of electron spin resonance spectra. It is suggested that the temperature-induced changes in the properties and activity of chloroplast membranes are part of a control mechanism for regulation of chloroplast development and photosynthesis by temperature.


Generation of electric current by chromatophores of Rhodospirillum rubrum and reconstitution of electrogenic function in subchromatophore pigment-protein complexes. Lipoprotein complexes, containing (1) bacteriochlorophyll reaction centers, (2) bacteriochlorophyll light-harvesting antenna or (3) both reaction centers and antenna, have been isolated from chromatophores of non-sulphur purple bacteria Rhodospirillum rubrum by detergent treatments. The method of reconstituting the proteoliposomes containing these complexes is described. Being associtated with planas azolectin membrane, ptoteoliposomes as well as intact chromatophores were found to generate a light-dependent transmembrane electric potential difference measured by Ag/AgC1 electrodes and voltmeter. The direction of the electric field inproteoliposomes can be regulated by the addition of antenna complexes to the reconstitution mixture. The reaction center complex proteoliposomes generate an electric field of a direction opposite to that in chromatophores, whereas proteoliposomes containing reaction center complexes and a sufficient amount of antenna complexes produce a potential difference as in chromatophores. ATP and inorganic pyrophosphate, besides light, were shown to be usable as energy sources for electric generation in chromatophores associated with planar membrane.


Glycine transport by hemolysed and restored pigeon red cells. Effects of a Donnan-induced electrical potential on entry and exit kinetics. The influence of a Donnan effect on the transport of glycine by hemolysed and restored pigeon red cells was examined. The Donnan effect was produced by replacing Cl- with 2,4-toluenedisulfonate or glutamate. The effects of the associated membrane potential and inside-outside pH difference on glycine entry and exit rates were examined. The effects of pH on entry and exit rates in the absence of a Donnan effect were also examined. In the absence of a Donnan effect, Na+-dependent glycine entry requires the protonated form of a group with a pKapp of 7.9 and the deprotonated form of another group with a pKapp of 6.8. Neither of these are required for exit but the deprotonated form of a group(s) with a pKapp of 6.2 is required. The pK 7.9 group and pK 6.2 group probably react with H+ at the inner face of the membrane and the pK 6.8 group probably reacts at the outer face. The V for glycine entry was determined for cells with their Cl- largely replaced by toluenedisulfonate and without such replacement. Between pH 6.1 and 7, the ratio of the respective V values, VT/VC1, was 1.5-1.7. VT/VC1 rose above pH 7 to near 4 at pH 8.3. At pH 6.9, with glutamate replacing cell Cl-, the analogous ratio (VGlu/VC1) was 1.7. The increase of VT/VC1 above pH 7 could be quantitatively accounted for by the increase in cell [H+]/medium [H+] caused by the Donnan effect together with the assumption that the pK 7.9 group reacts with H+ at the inner face of the membrane. When cell Cl- was replaced by toluenedisulfonate or glutamate there was a drop in the term in the glycine Km describing Na+ dependence of glycine entry. When cell Cl- was replaced by toluenedisulfonate therewas a rise in the Na+-independent term in the glycine entry Km. By replacing varying amounts of cell Cl- with either toluenedisulfonate or glutamate, plots were obtained of entry rates vs. the cell [Cl-]/ medium [Cl-] ratio consistent with the assumption that the Donnan-induced membrane potential acts on a "moving" charge. Glycine exit was only slightly accelerated by trans-toluenedisulfonate. The ratio, exit rate into toluenedisulfonate medium/exit rate into Cl- medium rose with decreasing pH. This rise could be accounted for by a Donnan-induced inside-outside pH difference which affects a pKapp 6.2 group reacting with internal H+. The observed influences of the Donnan effect on V (glycine entry), on both components of Km (glycine entry), on the shape of the plot of glycine entry rate vs. the cell [Cl-]/medium [Cl-] ratio and on glycine exit all fit the assumptions that when the empty porter reorients, one unit of negative charge accompanies it "across" the membrane and that no other steps involve charge movement. The properties of the system seem inconsistent with a translational ("ferry boat") mobile carrier.


Stimulatory effect of bleomycin on the synthesis of acidic glycosaminoglycans in cultured fibroblasts derived from rat carrageenin granuloma. Cultured fibroblasts derived from rat carrageenin granuloma were treated with bleomycin and the synthesis of hexosamine-containing substances was compared with that in control cells. Four day treatment with o.1 mug bleomycin/ml resulted in a significant increase of the production of these macromolecules by the cells, though DNA synthesis was remarkably inhibited at this dose of bleomycin. The stimulatory effect could be seen as early as the second day of bleomycin treatment, and was enhanced with increasing treatment time. Further fractionation of the hexosamine-containing substances revealed that synthesis of acidic glycosaminoglycans was more sensitive to bleomycin than that of glycoproteins, i.e., acidic glycosaminoglycans increased by 80% and glycoproteins by 53% after four day treatment with 0.1 mug bleomycin/ml. The increased components of acidic glycosaminoglycans included not only hyaluronic acid but also sulphated glycosaminoglycans. Collagen synthesis was increased by 23% by the same dose of bleomycin. N-Acetyl-beta-glucosaminidase, one of the degradation enzymes for acidic glycosaminoglycans released into the cultured medium, was decreased significantly by bleomycin.


Effect of ionophores on intralysosomal pH. 1. The effect of ionophores on the intralysosomal pH (as estimated from the distribution of a weak acid or base), on the distribution of 42K+ across the lysosomal membrane, and on the intralysosomal degradation of 125I-labelled bovine serum albumin has been studied. 2. Nigericin and X537A equilibrate both 42K+ and H+ across the lysosomal membrane. Gramicidin equilibrates H+ across the lysosomal membrane, this equilibration being more effective in a NaCl than in a KCl medium. Thus all three ionophores exhibit the same ion specificity as in other membranes. 3. The effect of the exchange-diffusion ionophores cannot be imitated by the combination of valinomycin with an uncoupler. Valinomycin by itself also has no effect. 4. X537A and nigericin inhibit the intralysosomal degradation of 125I-labelled albumin only when potassium is present. In a sucrose-containing medium no effect is found. Similar results were obtained with gramicidin. 5. These data suggest that the lysosomal membrane is impermeable to monovalent cations at 25 or 37 degrees C, and that the transport of protons is organised in such a way that electroneutrality is maintained.


Dynamics of fast axonal transport.

Dynamics of fast axonal transport. A phenomenological model of the process of fast axoplasmic transport is presented. The process was conceived of as occurring in two parts: (a) synthesis and storage of material in a cytoplasmic pool; (b) release from the pool and transport distally along the axon. Considering the fate of labeled proteins, the activity at points along the axon relfects events occurring earlier within the pool through the relationship: g(x,t) = const f(t - x/v); where g(x,t) represents axonal activity, f(t) the pool's activity, and v is the transport speed. Using the idea that when there is no further input of radioactivity into the pool its activity declines exponentially due to export of material to the axon. I generalized this concept to the case where activity enters and leaves the pool simultaneously. The model contains two parameters: the relative turnover rate of the pool, alpha, and T, an interval characteristic of the time of synthesis. From this model, the experimental data is unfolded and yields values for these parameters of alpha = 0.004 min-1 and T approximately 60 min.


Comparison of normal and chronic lymphocytic leukemia lymphocyte surface Ig determinants using peroxidase-labeled antibodies. II. quantification of light chain determinants in atypical lymphocytic leukemia. Five cases of atypical lymphocytic leukemia were investigated with regard to their membrane-associated light chains. Detection and quantitation of antigenic determinants were performed by means of peroxidase-labeled antibodies according to Avrameas et al. The cases studied had clinical and cytologic features in common: an active clinical course, marked splenomegaly, severe anemia and thrombocytopenia, little or no lymph node enlargement, and very high white blood counts with small mature lymphocytes and poorly differentiated lymphoid cells. Blood lymphocytes of all patients carried a single type of light chain, and 90%-100% of the cells were stained. The average number of antigenic sites per cell was 72,500 (range 40,000-97,500). These results differed from those previously found in typical CLL (mean value 9000) and approached the values of normal peripheral blood lymphocytes (90,000). The criteria investigated in this study could be of value for the diagnosis and prognosis of some atypical forms of lymphocytic leukemia.


[Significance of PAS-positive myeloblastic leukemia]. We refer to 48 adult patients suffering from acute myeloblastic leukemia, whose leukemic cells showed a typical cytochemical pattern: i.e. weak staining for peroxydase and with sudanblack B and lacking or only weak staining for nonspecific esterase. In 8 patients the leukemic myeloblasts additionally showed distinct granular staining for polysaccharides using the PAS-reaction. The therapeutic response, the remission rate and the survival time of these 8 cases have been compared to those of 40 patients, whose leukemic myeloblasts differed exclusively in the absence of granular PAS-positive materials. Out of the latter PAS-negative cases 7 patients (18%) went into a complete remission (M1, P1), 5 patients achieved partial remission (M2, P1-2), the 50%-survival time was 4.9 months. Out of the 8 granular PAS-positive cases 4 patients (50%) went into complete (M1, P1), and 1 reached partial remission (M2, P1-2). The 50%-survival time of the cases now lasts 10.1 months and three patients are still alive and in persisting complete remission 19, 12.5 and 12 months after diagnosis. These results suggest a better prognosis and an improved therapeutic response in those myeloblastic leukemias which additionally contain cytoplasmic granular PAS-positive materials.


Lead concentrations in soil and vegetation associated with highways of different traffic densities. Soil lead levels were found to decrease significantly in all traffic areas as distance from the highway increased. Areas of higher traffic volume had greater soil lead levels. Strong trends were shown in vegetation with areas of greater traffic density having greater lead levels than areas of lower traffic density. A decrease in lead levels was also noted in vegetation as distance from highway increased.


The use of alpha-adrenergic blockers in benign prostatic obstruction. As a result of previous in-vitro studies on the alpha-adrenergic receptor activity of the human prostate and prostatic capsule a trial was made of alpha-adrenergic blockers for the relief of obstructive prostatic symptoms. Considerable benefit was obtained in several groups of patients, as demonstrated either by the relief or prevention of complete retention, or by diminution in residual urine or improvement in urinary flow-rate recordings. A number of illustrative cases are described, and the indications for the use of this treatment are suggested. It is emphasised that this treatment provides symptomatic relief only, and in no sense purports to be a treatment of the enlarged prostate itself.


The results of surgical treatment for carcinoma of the rectum of St Mark's Hospital from 1948 to 1972. The surgical treatment of 3163 patients seen at St Mark's Hospital with a single adenocarcinoma of the rectum in the years 1948-72 is described and the results analysed. In 2948 patients (93-2 per cent) the primary tumour was removed. The operative mortality fell from 7-0 per cent in the years 1948-52 to 2-1 per cent in 1968-72. The proportion of restorative operations has risen steadily over the years to a level of 41-1 per cent in the years 1968-72. There were 2410 operation survivors in the years 1948-67. The crude 5-year survival rate in the whole group was 47-1 per cent (corrected figure 56-7 per cent), and 56-6 per cent (corrected figure 68-4 per cent) for the 1931 survivors of radical operations. Comparison of results for patients surviving radical synchronous combined excision and radical anterior resection shows a significant difference in the two groups: in the former the crude 5-year survival rate was 52-7 per cent (corrected figure 63-8 per cent), and in the latter group the respective figures were 66-7 and 79-4 per cent. A higher proportion of Dukes' A and B cases and of low grade tumours are shown as the pathological background to the more favourable prognosis for patients surviving radical anterior resection.


Spontaneous rupture of liver tumours.

Spontaneous rupture of liver tumours. Five geriatric patients with spontaneous rupture of the liver caused by hepatic malignancies were operated upon without mortality. Liver resection seems to be the treatment of choice, but in selected cases with unresectable tumours ligation of the hepatic artery alone is preferable to an attempt to control haemorrhage by packing and suture.


Selective uptake and retrograde axonal transport of dopamine-beta-hydroxylase antibodies in peripheral adrenergic neurons. In the present experiments the uptake and retrograde axonal transport of antibodies to dopamine beta-hydroxylase (DBH) in adrenergic neurons was studied. When partially purified labelled antibodies to DBH were injected unilaterally into the vicinity of the adrenergic nerve terminals in the iris, radioactive substances accumulated preferentially in the superior cervical ganglia of the injected. By SDS (sodium dodecyl sulfate) gel electrophoresis and immunoprecipitation it could be shown that the accumulated radioactivity in the superior cervical ganglion represented antibodies to DBH. This retrograde accumulation was greatly reduced by colchicine, axotomy or destruction of the adrenergic nerve terminals by 6-hydroxydopamine. The rate of retrograde transport was the same as that of nerve growth factor (NGF) and tetanus toxin in sympathetic neurons. The retrograde transport of antibodies was confined to sympathetic neurons and could not be detect in either sensory or motor neurons.


[Increase of resistance to Pseudomonas aeruginosa infection in mice caused by Staphylococcus aureus]. In our study of opportunistic pathogens, we have some indication that Staphylococcus aureus can increase resistance in mice against Pseudomonas aeruginosa. Intraperitoneal injections of sublethal doses of S. aureus had a protective effect in mice against lethal doses of P. aeruginosa, more so if living and coagulase-positive S. aureus strains were injected. This protective effect was obtained both with laboratory and freshly isolated hospital strains. The interval between these infections can be extended from 2 h up to 1 week and it is still possible to observe the resistance phenomenon. The increased resistance was accompanied by a decrease in viable units of P. aeruginosa in the peritoneal cavity of mice 6 h after the injection of this species. There was no protection by S. aureus against Candida albicans in similar experimental conditions. These observations indicate that intermicrobial ecology, understood here as the previous presence of another species in a host, may be a significant factor in the resistance to infection with opportunistic pathogens such as P. aeruginosa.


[Bidimensional immunoelectrophoretic study of the antigenic composition of the membrane in various mycoplasma strains]. Membrane antigenic composition of Acheloplasma laidlawii PG9, A. granularum BTS-39, and Mycoplasma fermentans PG 18(G) was determined by means of bidimensional immunoelectrophoresis in the presence of sodium desoxycholate 0.5%. Depending upon the mycoplasma species from which membranes were obtained, 7 to 15 antigens were evidenced. Using sodium desoxycholate presents the advantage over non-ionic detergents to dissolve better the mycoplasmic membrane antigenic complexes. A comparative study of five strains belonging to the above-noted species confirms the serological heterogeneity of the Mycoplasmateles order and shows variability at the membrane antigenic composition level of Acheloplasma laidlawii.


Isolation and characterization of phenotypes of mycobacteria. Several strains of mycobacteria grown as surface pellicle on liquid Sauton's medium under semianaerobic conditions dissociated into three phenotypes: phenotypes 1, 2, and 3. Only phenotype 1 could be obtained in a pure state. None of these phenotypes was found to be stable: they convert from one into another and all revert to the parental strain when replaced in their usual aerobic cultural conditions. Comparative studies of phenotypes 1 and 3 have shown that significant differences exist in their physiological behaviour and antigenic composition.


Uptake of L-proline by Histoplasma capsulatum. The uptake and incorporation of L-proline by yeast cells of the dimorphic zoopathogen Histoplasma capsulatum were studied. The amino acid was assimilated in at least two ways: by an active transport system with a Km of 1.7 X 10(-5) M and by simple diffusion. The active transport system was sterospecific and severely restricted to neutral aliphatic side-chain amino acids. Certain analogues inhibited L-proline uptake and prevented incorporation of the amino acid into cellular constituents. The inhibition of L-proline uptake by L-leucine was competitive. Since L-leucine and L-proline are seemingly transported by a system with similar characteristics, must be concluded, as originally postulated, that the buckled ring of L-proline, in solution, acts as an aliphatic side chain and that this cyclic amino acid is transported by a system more or less specific for amino acids with neutral aliphatic side chains.


A light and electron microscope study of changes occurring at the cut ends following section of the dorsal roots of rat spinal nerves. Rat dorsal spinal nerve roots were cut; 20 h later the axons in the vicinity of the cut were examined by light and electron microscopy. The changes in the cut tip distant from the ganglion were largely degenerative. On the ganglionic side of the cut a cap of free unmyelinated sprouts was formed. These sprouts contained clear and dense-core vesicles 40-150 nm in diameter, smooth endoplasmic reticulum and mitochondria. Some of the unmyelinated sprouts were extensions of myelinated axons, others arose from myelinated axons by lateral budding. In both myelinated and non-myelinated axons there was an accumulation of mitochondria, tubulo-vesicular smooth endoplasmic reticulum and large and small dense-core vesicles for a distance of approximately 500 mum behind the tip. Dense-core vesicles were more common in non-myelinated axons than in their myelinated counterparts. In areas of intense accumulation the non-myelinated fibres were grossly swollen and distorted. The myelinated axons and some of the sprouts contained an unusual type of mitochondrion. The similarity between these sprouts and pre-synaptic terminals is discussed.


Amplification and characterization of a beta-globin gene synthesized in vitro. Full-length, double-stranded globin DNA was synthesized in vitro starting from rabbit globin mRNA. Several restriction endonuclease cleavage sites with known recognition sequences were mapped on this DNA as a means of assessing the accuracy of in vitro synthesis. By comparing this map with the nucleotide sequences known or predicted from the amino acid sequences of alpha-and beta-chain rabbit hemoglobin, it was possible to show that the synthetic globin DNA is a faithful copy of beta-globin mRNA. Amplification of the synthetic globin DNA  was achieved by inserting the molecule into the plasmid PMB9 using the poly(dA)-(dT) joining procedure, and transforming E. coli with the hybrid DNA. Transformants carrying beta-globin DNA were identified by colony hybridization using purified 125I-beta-mRNA probe. Comparison of the restriction maps of the synthetic and inserted globin DNAs showed that the entire synthetic globin DNA molecule was amplified without sequence rearrangements. Both the synthetic and the cloned DNA include the entire coding sequence of the beta-globin gene plus a substantial portion of the untranslated regions flanking the structural gene.


Macrophage plasminogen activator: modulation of enzyme production by anti-inflammatory steroids, mitotic inhibitors, and cyclic nucleotides. Plasminogen activator production by cultured mouse peritoneal macrophages can be modulated in vitro by low concentrations of various pharmacologically active molecules. Glucocorticoid hormones and their synthetic derivatives, as well as cholera toxin, colchicine, and vinblastine markedly inhibit production of this enzyme without affecting other important macrophage functions. The effect of glucocorticoids is of particular interest, both because their relative in vivo anti-inflammatory potencies correlate exactly with their effect on plasminogen activator production in culture and because this effect occurs at near physiological concentrations. In view of the correlations established in other systems between plasminogen activator production and cell migration, we have also examined the age of the macrophages in thioglycollate-induced exudates. Confirming the results of Van Furth and Cohn (1968), we have found that the majority of these cells are young, having recently  replicated and arrived in the peritoneal cavity. Using a fibrinagar overlay technique which allowed us to determine the production of plasminogen activator by individual cells. we have found that the majority of these cells produce the enzyme. The potential roles of plasminogen activator in monocyte migration and the relationship of this enzyme to the anti-inflammatory effect of gluccorticoids are correlated and emphasized.


The similarity of changes in segmental contraction patterns induced by postextrasystolic potentiation and nitroglycerin. Despite a fundamental difference in their underlying mechanisms, both postextrasystolic potentiation (PESP) and administration of nitroglycerin (TNG) have been utilized to predict reversibility of abnormal segmental wall motion in patients with ischemic heart disease. To determine whether these interventions induce the same changes in segmental contraction pattern, we analyzed biplane ventriculograms of 14 patients who had an adequately visualized PESP beat on a basal ventriculogram as well as a post-TNG ventriculogram. Four segments in each plane were defined and the area ejection fraction of each segment was calculated for a basal sinus, PESP, and post-TNG beat. To correct for global differences in the response to PESP and TNG, we normalized each segmental ejection fraction (NSEF) by the ventricular ejection fraction for that beat and then compared the differences in NSEF from the basal value after PESP and TNG. Eleven patients demonstrated similar responses to both interventions. The three patients whose responses were discordant had elevated or unchanged left ventricular systolic or end-diastolic pressures at the time of the TNG ventriculogram. Our data suggest that, provided these pressures are lower than basal values at the time of the TNG ventriculogram, PESP and TNG will induce similar changes in segmental contraction patterns. Seven patients with similar responses had a PESP beat on their post-TNG ventriculogram. Changes in NSEF after PESP+TNG were identical to those after either intervention. This implies that the combination of interventions does not induce further changes in segmental contraction pattern beyond that produced by either intervention alone.


Serum cholesterol and bile acid in primary hepatoma. The close relationship between serum levels of cholesterol and bile acid has been confirmed in 46 patients with primary hepatoma. Serum levels of cholesterol and bile acid are roughly correlated with serum alpha-fetoprotein concentration. Because the relationship between serum cholesterol and bile acid did not exist in common hepatocellular diseases, the results suggest a peculiar sterol metabolism occurring in human hepatoma.


[Determination of alpha-1-foetoprotein in human serum by electroimmuno-diffusion on agarose (author's transl)]. The electroimmunodiffusion on agarose has been applied for the determination of AFP in human serum. The proposed method can be used for concentrations between 0.6 and 50 mg/l; the reproducibility was judged to be acceptable. For concentrations higher than 10 mg/l, a good correlation was observed between electroimmunodiffusion and radial immunodiffusion.


[Biochemical modifications in a case of analbuminemia (author's transl)]. A new case of analbuminemia is described for a six month old child of Algerian origin. The serum albumin concentration was 64 mg/l and its immunochemical action was identical to that of normal albumin. The system reacted by an increase of the synthesis of globulins. For the subject, the alpha1-antitrypsin, ceruloplasmin, haptoglobin, alpha2-macroglobulin, transferrin and immunoglobulins M contents were three times higher than the standard figures. However, it was possible to show that the presence of free bilirubin independent from proteins could be detected at a concentration of 17 mumol/l.


An unusual alkaline phosphatase isoenzyme in a patient with concomitant marked elevation of acid phosphatase without clinical evidence of prostatic carcinoma. An 81-year-old male with congestive heart failure and prostatic hypertrophy was found to have markedly elevated serum acid phosphatase (EC 3.1.3.2) and moderately elevated serum alkaline phosphatase (EC 3.1.3.1). Alkaline phosphatase isoenzyme analysis was performed to determine the organ source of the enzyme. There was an unsual slow-migrating alkaline phosphatase isoenzyme band in the serum of this patient suggestive of hepatic origin by a variety of biochemical tests.


Alpha-fetoprotein-like activity in sera from patients with malignant and non-malignant disease and healthy individuals. A new method, radio-crossed immunoelectrophoresis, demonstrates alpha-fetoprotein (AFP) in sera with a sensitivity of 1 mug/1. By this method AFP with alpha mobility was not found in sera from healthy individuals, patients with chronic active hepatitis and cirrhosis, primary biliary cirrhosis, secondary liver cancer and cystic fibrosis. In some of the sera, AFP was elevated when measured by conventional radioimmunoassay method and the sera contained an AFP-like substance with gamma mobility when analyzed by radio-crossed immunoelectrophoresis. The nature of this gamma substance is still obscure and needs further investigation.


Dissociation between TSH and prolactin dynamics in treated thyrotoxicosis. TSH and prolactin secretory patterns in thyroid disease have generally been reported as concordant. We studied TSH and prolactin responses to TRH infusion (500 mug) in euthyroid individuals previously treated for thyrotoxicosis with 131I or antithyroid drugs. The 131I-treated group (seven men, twenty women) had been clinically and biochemically euthyroid (normal serum thyroxine and triiodothyronine levels) for 6 months to 4-5 years (kappa +/- SD = 17-1 +/- 4-1 months). Based on maximal TSH increment (deltaTSH), three patient groups were identified: Group 1 [normal deltaTSH, n = 6]: delta prolactin was normal in two, blunted in one and exaggerated in three. Group 2 (exaggerated TSH response, n = 8): delta prolactin was normal in two, blunted in one and exaggerated in five. Group 3 (TSH nonresponders, n = 13): delta prolactin was normal in five, blunted in three, and exaggerated in five. Eleven patients (three men, eight women) were studied after 6 months antithyroid-drug treatment. All were clinically and biochemically euthyroid. All but one showed a blunted TRH-TSH response. All three men showed an exaggerated delta prolactin as did four of eight women. Three women showed a blunted delta prolactin and in one, delta prolactin was normal. Thus, TRH-induced TSH and prolactin response patterns in treated thyrotoxicosis are not uniformly concordant, and, while a blunted or absent TSH response commonly persists long after euthyroidism has been restored, this is most frequently accompanied by a normal or exaggerated prolactin response.


Tissue and hepatic subcellular distribution of liposomes containing bleomycin after intravenous administration to patients with neoplasms. 1. Liposomes containing 111In-labelled bleomycin were injected intravenously into two patients. One patient had a hepatoma and the other had secondary adenocarcinomatous deposits in the liver. 2. The tissue distribution of 111In was determined by whole-body scanning and by measurement of the radioactivity in organs at autopsy. 3. Scans in vivo and post-mortem measurement of radioactivity indicated that liposomes accumulate predominantly in the liver, but that there is no selective uptake of liposomes by the malignant tissue. 4. The subcellular distribution of radioactivity in the liver was measured 90 min after injection by fractionation of percutaneous liver biopsies on sucrose density gradients. 5. Radioactivity within the liver was concentrated in lysosomes. 6. Electron microscopy of tissue obtained before the administration of liposomes revealed particles morphologically indistinguishable from liposomes in hepatoma cells and hepatocytes.


A test of granulocyte membrane integrity and phagocytic function. An assay of granulocyte viability has been developed which yields information about rwo important cell parameters, cell membrane integrity and phagocytic activity. The assay is based on the fact that only live cells can accumulate fluorescein, which is enzymatically split from the nonfluorescent substrate fluorescein diacetate. Dead cells, on the other hand, become permeable to the fluorescent red dye ethidium bromide. When cells are exposed first to opsonized zymosan particles, which they can phagocytize, then to a combination of these fluorescent dyes, one can distinguish microscopically between dead cells with fluorescent red nuclei, live cells which fluoresce green, and live cells with phagocytic function which are swollen with the pink zymosan particles in a green fluorescing cytoplasm. This assay takes 20--30 min and can be used to distinguish different degrees of cellular damage after cryopreservation.


Cryoprotectant-treated myocardium evaluation. Whole excis ed rat hearts were treated with 5, 10, or 15% (v/v) dimethyl sulfoxide/glycerol and then some were frozen in liquid nitrogen while the balance remained unfrozen. Freezing and thawing rates were approximately 30degreesC/min. Ventricular tissue was examined for histological damage, glycogen depletion, and enzyme sites, using histological, histochemical, and electron microscopy techniques. Early signs of cellular degeneration and ischemia were observed in all unfrozen groups; depletion of glycogen reserves, fuchsinophilic staining, vacuolization and granulation of the sarcoplasm were noted. Results from frozen groups were similar, but ultrastructural damage was more severe and extensive. Alkaline phosphatase and succinic dehydrogenase sites were abundant in unfrozen specimens and absent or markedly reduced in frozen specimens which also exhibited widespread nonspecific staining throughout intercellular spaces.


[Intraoperative vital staining of insulinoma using toluidine blue-o]. A case is reported in which in vivo staining of insuloma cells was performed intraoperatively, using Toluidine Blue-O. The surgical problems of islet-cell tumors are discussed with special regard to the indication of in vivo staining as a definite method of intraoperative identification of these tumors.


Mechanisms of chromosome banding. IX. Are variations in DNA base composition adequate to account for quinacrine, Hoechst 33258 and daunomycin banding? Prior studies on subfractions of mouse and Kangaroo rat DNA have suggested that variations in base concentration within a given genome may not be great enough to account for Q-banding. To examine this with another species, calf DNA was subfractionated by CsCl ultracentrifugation into GC-rich satellites and the main band DNA was further fractionated into AT-rich, intermediate and GC-rich portions. The effect of varying concentrations of these DNAs on quinacrine and Hoechst 33258 fluorescence was examined. Although with both compounds there was less fluorescence in the presence of the GC-rich satellites than main band fractions, these results per se did not answer the question of whether the variation in base composition alone was adequate to account for chromosome banding. To answer this the fluorescence observed in the presence of DNA of a given base composition was related to the fluorescence observed in the presence of DNA of 40% GC content (F/F40). This allowed the derivation of a term B which indicated the relative change in fluorescence per 1% change in base composition of DNA. To determine the percent change in fluorescence observed in Q-banding, the photoelectric recordings of Caspersson et al. (1971) were used. From these data we conclude: 1. Quinacrine is twice as sensitive to changes in base composition as Hoechst 33258. 2. Variation in the base content of DNA along the base content of DNA along the chromosome is sufficient to account for most Q-banding, except possibly for some of the extremes of quinacrine fluorescence. This was further examined with daunomycin. Even though daunomycin gives good fluorescent banding, DNAs varying in base composition from 100 to 40% GC content all resulted in the same relative fluorescence of 0.03. However, in the presence of poly (dA-dT) the relative fluorescence was 0.85, indicating a great sensitivity to very AT-rich DNA. This suggests that with daunomycin and possibly other fluorochromes, stretches of very AT-rich DNA may be more important in fluorescent banding than simple variation in mean base composition.


The transfer of J blood-group activity from J-containing bovine serum to J-negative erythrocytes. The J blood-group activity of bovine serum is contained both in a lipid and in a nonlipid fraction. This is also true for calf serum. It demonstrated that the J determinant is transferred from a serum protein onto the erythrocyte membrane by incubation in vitro. Even though the donor of J activity is a lipid-free serum protein (probably a glycoprotein), the transferred J activity is detectable only in the lipid fraction of erythrocytes. Thus, the J determinant (probably a carbohydrate unit) must have been detached from a serum glycoprotein and transferred to a lipidic receptor (probably a glycosphingolipid) at the erythrocyte membrane. It is suggested than an enzyme system located in or at the erythrocyte membrane is responsible for the transfer of J substance. The transfer of J substance is inhibited by a polar lipid present in bovine serum.


Late results following cryosurgery of the prostate in 111 high-risk patients. The so-called cryoprostatectomy was performed on 111 high-risk patients from November 1970 to December 1975 (60-month period). 86 patients (78.5%) had an adenoma and 25 patients (21.5%) a carcinoma of the prostate. During this period of time 1,085 more patients with adenoma or carcinoma were treated by open prostatectomy or TUR. Experience has shown that cryoprostatectomy should be performed only on high-risk patients whose physical condition excludes operation and the only alternative would be continuous urethral catheter drainage. Any further indication must be rejected.


The avascular nephrotomy.

The avascular nephrotomy. A simple and innocuous method for colouring the renal cortex is proposed. It makes visible, during the operation, the line of demarcation between different arterial areas and allows the performance of nephrotomies with minimal trauma to the renal parenchyma and a minimum of subsequent renal destruction by ischaemia. It is indicated as a complement to renal hypothermia, for the removal of complicated and extensive lithiasis; it can also be helpful in easier cases when hypothermia in not required.


Uretero-colic anastomosis by the 'cuff' or 'flap' procedure--Goodwin's modified technique: preliminary report. Goodwin's procedure of uretero-colic anastomosis under direct vision, performed completely extraperitoneally by the endo-intestinal route marks a considerable progress when compared with the different variations of Coffey's operation. This technique may safely be applied after palliative cystectomy for proliferating bladder cancer if the anastomosis is performed at some distance from the common site of pelvic recurrence, and is provided with a system against back flow. In this regard the 'cuff' process seems quite satisfactory.


Cell-mediated immunity in patients with renal adenocarcinoma. The lymphocytotoxicity test was used to study the existence of immunologically active lymphocyte reactions in patients with renal carcinoma at different stages. In 9 patients with an advanced renal carcioma (stage IV) a low lymphocytotoxicity was found, while the examination of 7 patients with stage I and II renal tumours showed a high value of the lymphocytotoxicity test. The clinical importance of the lymphocytotoxicity test is discussed with special interest in a possible immunotherapy.


Iglesias resectoscope with simultaneous irrigation, suction and low intravesical pressure. A resectoscope with continuous irrigation, suction and low intravesical pressure has been described. Advantages of this instrument include: no interruption, better endoscopic vision by a continuous clear inflow of more than 500 ml/min at 70 cm pressure, less bleeding, no air bubbles, constant intravesical pressure less than 10 mm Hg during TUR, permitting the absorption of the irrigant, shorter operating time, easier teaching and no more wet floor and wet surgeon. Since the entire amount of irrigating fluid is collected, blood loss can be calculated and the amount of absorption determined.


Placebo-controlled double-blind study in human benign obstructive prostatic hypertrophy with flutamide. A double placebo-controlled study using the anti-androgen non-steroidal compound Flutamide in 32 patients with benign enlargement of the prostate is reported. A statistical analysis of the results showed no difference between the group treated with Flutamide and the group receiving the placebo, except in as far as 'the size' of the prostate was concerned. We observed a significant reduction of the latter in patients receiving Flutamide. The different problems produced by the choice and the measure of parameters in this type of investigation have been discussed.


The expression of human beta2-microglobulin on human spermatozoa. The expression of human beta2-microglobulin was detected on human spermatozoa from 7 donors using a cytotoxicity assay and indirect immunofluorescence. Inhibition experiments using purified beta2-microglobulin demonstrated the specificities of these reactions. We were able to localize beta2-microglobulin on human spermatozoa on the post-acrosomal region at a similar location to those described for HL-A antigen and for early embryonic antigen +t12, but also at the end of the intermediary piece and the beginning of the tail.


Beta2-microglobulin on the cell surface. Specificity of inhibition of the mixed leukocyte reaction and mitogenic properties of antibodies against the two HLA antigen polypeptide chains. Intact antibodies, and F(ab')2, and Fab' fragments thereof directed against beta2-microglobulin (beta2m) and the HLA alloantigenic polypeptide chain impede the mixed leukocyte culture reaction (MLR). The Fab' fragments are effective only when bound to the responder cell population. The kinetics of their inhibiting action suggest that they interfere with the recognition phase of the MLR. Thus, it seems possible that beta2m may be part of or closely associated with those structures on the responding cells which recognize the stimulatory alloantigens. Antibodies against beta2m or the other HLA antigen polypeptide chain are mitogenic for peripheral blood lymphocytes. The antibodies against beta2m caused greater stimulation than those against the HLA alloantigenic polypeptide chain. Only bivalent antibodies against beta2m were mitogenic and promoted blast transformation of 1 to 2% of the lymphocytes. Although only B lymphocytes were triggered, the mitogenic response of cultures containing various proportions of B and T cells suggests that cooperation of the two cell types is necessary to obtain optimal mitogenic stimulation.


The length of parallel fibers in the cat cerebellar cortex. An experimental light and electron microscopic study. Experimental light and electron microscopic studies were carried out to determine the length of parallel fibers in the cat cerebellar cortex. Using a fine surgical knife, vermal and hemispheral folia were cut perpendicular to their long axis. The animals were sacrificed 1-10 days after the operation. Sections of the transected folia were then stained with a Fink-Heimer procedure. The resulting degeneration appeared as fine dots that extended lateral to the lesion, as predictable from the course of the parallel fibers. Densitometer readings indicate that the density of degeneration declines gradually lateral to the lesion. The specificity of the silver impregnation was checked by processing silver stained sections for electron microscopy. This confirmed the location of the silver precipitate on degenerating parallel fibers. The pattern of parallel fiber degeneration in the molecular layer has a trapezoidal configuration centered on the lesion. The shorter parallel fibers are located at the base of the molecular layer and extend for 5 mm. The parallel fibers become progressively longer as they approach the pial surface where they attain a maximum length of 7 mm. Our studies suggest that in folia longer than 7 mm parallel fibers are 6 mm long on the average. In addition, it was determined on Golgi sections that the average center-to-center distance between en passant boutons of individual parallel fibers is 5.2 mum. The data indicate that an average parallel fiber, 6 mm long, forming approximately 1100 boutons, may synapse with each Purkinje dendritic tree it traverses.


Concanavalin A receptors in the plasma membrane of rat liver cells: comparative electron microscopic studies on normal cells and on cells in vivo transformed by diethylnitrosamine. The Concanavalin A receptors at the cell surface of normal rat liver cells and of those in vivo transformed by diethylnitrosamine were comparatively studied by electron microscopic cytochemistry. Besides different agglutinability of the cells a variable surface staining of the cells by the Con A-peroxidase reaction (BERNHARD and AVRAMEAS 1971) was observed. After performance of the cytochemical reaction on living normal rat liver cells in situ a continuous cell surface staining was seen. In transformed rat liver cells a marked tendency for patchy distribution of the Con A label at the cell surface occurred. Furthermore, internalisation of Con A-peroxidase labeled plasma membrane segments was visible in the transformed cells. A similar variable labeling by Con A-peroxidase reaction occured also in the "basal" plasma membrane of normal and transformed rat liver cells. The results are discussed with respect to the importance concerning the mobility of lectin receptors and membrane stability.


Technique for the visualization of exchange aberrations in human chromosomes. A technique for the visualization of chromatid exchange in human chromosomes is described. Additon of the base analogue 5-bromodeoxyuridine (BUdR) into PHA stimulated lymphocyte cultures and subsequent staining with the benzimidazol compound "33258 Hoechst", and Giemsa, results in differential fluorescence or staining of sister chromatids in second division metaphases. This technique allows an accurate scoring of sister chromatid exchanges, and could be used in the evaluation of the effect of external agents on the induction of previously unrecognized chromatid exchanges in human chromosomes.


Correlation of light and electron micrographs of human metaphase chromosomes after incorporation of BUdR and staining with "33258 Hoechst" and Giemsa. Stained (Giemsa, "33258 Hoechst"1) and "33258 Hoechst" + Giemsa) and unstained metaphase chromosomes from human peripheral lymphocytes, after two rounds of replication in the presence of 5-bromodeoxyuridine (BUdR), have been prepared for electron microscopy. There is a positive correlation between light and electron micrographs. The same differential contrast on electron micrographs has been obtained whether the preparations have been stained or not. We attribute this differential contrast primarily to the lesser condensation of the bifilarly substituted chromatid.