Immune complex receptors on cell surface. I. Ultrastructural demonstration of macrophages. A method is described for ultrastructural localization of immune complex receptors on the surface of viable peritoneal exudate cells. The technique entails incubation with a soluble complex of horseradish peroxidase (HRP) and specific antibody to HRP at 4 degrees C followed by exposure to diaminobenzidine and processing for electron microscopy. The bound immune complexes were evident as focal deposits of HRP reaction product, adhering closely to the external surface of macrophages with an uninterrupted periodicity varying between 30 and 120 nm. Following incubation with an insoluble immune complex containing a higher proportion of antibody, receptor sites stained frequently, but large aggregates adhered to the cells. Rinsing cells after staining with soluble complexes partially displaced the bound immune complexes. Fixation prior to exposure to immune complexes largely eliminated the binding capacity of the immune complex receptors.


Bence Jones proteins and light chains of immunoglobulins. XIII. Effect of elastase-like and chymotrypsin-like neutral proteases derived from human granulocytes on Bence Jones proteins. Bence Jones proteins can be cleaved specifically by several types of endopeptidases into fragments corresponding to the amino-terminal, variant (VL) portion and to the carboxyl-terminal, constant (CL) portion of the light polypeptide chain. Two types of neutral proteases, designated elastase-like (ELP) and chymotrypsin-like (CLP), have been isolated and purified from human polymorphonuclear leukocytes. Because these proteases have defined proteolytic activity under physiologic conditions for several types of human proteins, we investigated their effect on human Bence Jones proteins. Incubation of kappa-type or lambda-type Bence Jones proteins with ELP or CLP under appropriate conditions resulted in cleavage of both types of light chains as evident by immunochemical and electrophoretic analyses. Treatment with ELP or CLP of one kappa Bence Jones protein resulted in the formation of a single component that had antigenic and electrophoretic properties similar to the VL fragment derived from pepsin digestion of the native protein. No component corresponding to the CL could be detected immunochemically or electrophoretically. Studies of isolated pepsin-labile (37 degrees C) and pepsin-stable (55 degrees C) CL fragments demonstrated the marked susceptibility of the carboxyl-terminal half of the light chain to proteolysis by the leukocyte-derived neutral proteases. Incubation with ELP of three other kappa Bence Jones proteins and three reduced-alkylated lambda Bence Jones proteins resulted, in each case, in the formation of a homogeneous component which was electrophoretically and immunochemically distinct from the pepsin-derived VL fragment. An identical component could also be formed by incubating a pepsin-derived VL fragment with ELP. In the ELP-treated samples, no CL-related material was detected electrophoretically or immunochemically with antisera possessing specificity for CL antigenic determinants present on the unfolded light polypeptide chain or on the isolated CL. The component formed by ELP or CLP treatment of certain Bence Jones proteins thus appears to be VL-related, but lacks the idiotypic antigenic determinant present on the native protein. In this respect, these neutral protease-derived light chain components are similar to the amyloid-like VL fragments generated in vitro from certain endopeptidase-treated Bence Jones proteins.


Histamine release from cultured human basophils: lack of histamine resynthesis after antigenic release. Human peripheral basophils can be maintained in cultured for up to 72 hr. These cells retain their functional integrity as judged by total histamine content and by their ability to release histamine by an IgE-mediated reaction in response to a specific antigen challenge. Cells cultured after suboptimal antigenic challenge could be activated to release histamine upon the addition of antigen. In contrast, cells culture after supra-optimal antigenic challenge did not fully recover their ability to release histamine even after 24 hr. With a variety of culture conditions, it was impossible to demonstrate any net synthesis of histamine in cells. Cells cultured after depletion of their stores by reaction with antigen did not show any net histamine formation. The experiments suggest that the basophil in peripheral blood is functionally an end-stage cell and can participate in the histamine release reaction only once.


Modulation of cyclic AMP in purified rat mast cells. III. Studies on the effects of concanavalin A and anti-IgE on cyclic AMP concentrations during histamine release. Changes in rat mast cell cyclic adenosine 3',5' monophosphate (cAMP) concentrations during stimulation of histamine release by concanavalin A (con A) and anti-IgE were studied. Con A caused an increase in cAMP with a mean peak level at 20 sec of 232% of control (range 164% to 365%). Con A-stimulated cells demonstrated falls toward control levels after 20 sec, but generally remained above control for at least 5 min. By 10 min cAMP had returned to control values. The con A effect on cAMP occurred in the absence of phosphatidyl serine but was markedly inhibited by 5 mM alpha-methyl-D-mannose. Anti-IgE induced a less marked increase in cAMP (157% of control, range 110% to 540% of control) which reached a peak at 20 sec. Two monospecific goat anti-rat myeloma IgE antisera induced similar changes in cAMP whereas normal goat IgG had no effect. These peak values were followed by a rapid decrease in cAMP. Within 2 min the cAMP content of anti-IgE stimulated cells had fallen to levels well below control and remained below control levels from 45 sec to over 15 min. Histamine release in both systems began after the peak cAMP levels, during the period of rapid destruction of cAMP.


Interaction of C-reactive protein with lymphocytes and monocytes: complement-dependent adherence and phagocytosis. The serum constituent C-reactive protein (CRP), which activates the classical complement (C) pathway when reacting with its substrates, was examined for its ability to mediate reactions of opsonic adherence and phagocytosis. Erythrocytes coated with C-polysaccharide (CPS) and reacted with CRP (E. CPS-CRP) failed to adhere to B cells and displayed only minimal adherence to monocytes. However, upon the addition of absorbed C or purified C components these cells were found to possess the cleavage products C4b and C3b, which in turn resulted in attachment of these cells to both human B lymphocytes and peripheral blood monocytes. E. CPS-CRP treated with C in the absence of antibody were readily phagocytosized by glass-adherent human monocytes. The phagocytosis of E. CPS-CRP-C was not only mediated by CRP but also required the presence of CRP on the surface of the red cells. The extent of ingestion was proportional to the amount of CRP on the red cell intermediate and was reduced by blocking monocyte receptors with aggregated human gamma-globulin (HGG) at concentrations which did not impair the uptake of other particles. The mediation by CRP of reactions of opsonic adherence and phagocytosis as outlined in these studies points to a significant role for CRP in reactions of host defense and inflammation.


Studies on human brain-thymus cross-reactive antigens. Antigens shared by human brain and thymocytes and by human and mouse tissues were studied with rabbit anti-human thymocyte antiserum (RAHT). It was found that cytotoxicity of RAHT serum against mouse thymus cells was not absorbed by mouse liver or bone marrow cells. Human brain and thymus cells completely absorbed the anti-thymocyte activity from this antiserum. It was suggested that human brain had antigenic determinants identical or very similar to those found on human thymocytes. Activity of RAHT antiserum against mouse thymus cells was completely removed by an absorption of mouse brain and thymocytes. These results demonstrated that there were shared antigenic determinants between human and mouse tissues.


Synthetic glycolipid adjuvants.

Synthetic glycolipid adjuvants. In the course of the organic synthesis of model compounds similar in some features to the lipid moiety of endotoxic lipopolysaccharide (LPS), Nacylated-D-glucosamine derivatives were prepared. One of these, N-palmitoyl-D-glucosamine, has been previously found to be mitogenic for athymic nude mouse B cells. This and other N-acylated homologs were tested for adjuvant activity in the immune response to human gamma-globulin (HGG) and sheep red blood cells (SRBC) in mice. Comparable or superior enhancement of the immune response was obtained for these glycolipids when compared to LPS in assays measuring anti-SRBC or HGG hemagglutinin titers. In the determination of hemolytic plaque formation, considerable adjuvant effect was shown by the lauroyl derivative, and less but still significant enhancement was achieved by the N-palmitoyl-D-glucosamine. In the rosette formation assay, in addition to the above two glycolipids, N-oleyl-D-glucosamine showed good adjuvant effect. In the latter two assays, the LPS was a superior adjuvant as compared to the synthetic glycolipids. The radiation protective effect of some of the better synthetic adjuvants was also investigated in mice. It was found that although LPS was more effective in this assay, the N-myristoyl-D-glucosamine and N-decanoyl-D-glucosamine compounds gave a definite protection, since up to 40% of the lethally irradiated (700 R) mice survived.


Influenza A virus interaction with murine lymphocytes. I. The influence of influenza virus A/Japan 305 (H2N2) on the pattern of migration of recirculating lymphocytes. The effect of influenza virus A/Japan 305 (H2N2) on the path of migration of recirculating lymphocytes has been studied. 51Cr-labeled rat thoracic duct lymphocytes (TDL) were incubated with virus at 37 degrees C for 1 hr and then infused i.v. into syngeneic recipients which were killed 1 hr later. Virus-treated TDL accumulated in the liver and their recovery in lymph nodes and spleen was severely reduced. Changes in lymphocytes induced by virus developed rapidly and were evident after incubation for only 15 min. UV-irradiated virus altered the pattern of lymphocyte localization but attachment of heat-inactivated virus to lymphocytes in vitro had no effect on their distribution in vivo. Evidence was obtained that some virus-treated TDL, initially sequestered in the liver, subsequently recovered their ability to circulate normally. Recovery was not complete and a population of cells failed to regain their ability to home into lymph nodes. Evidence is also presented demonstrating that influenza virus affected the homing properties of both T and B cells. It is suggested that aberrations in lymphocyte homing were mediated by the viral neuraminidase which induces changes in the cell membrane leading to their accumulation in the liver.


Suppression of reaging antibody formation. IV. Suppression of reaginic antibodies to penicillin in the mouse. Reaginic antibodies to the benzylpenicilloyl determinant (BPO) and ovalbumin (OA) were induced readily in B6D2F1 mice by a single i.p. injection of either 1 or 10 mug of BPO4-OA suspended with 1 mg of Al(OH)3 in 0.5 ml of saline. Administration of conjugates consisting of the hapten coupled to the isologous, nonimmunogenic murine gamma-globulins (MgammaG), i.e., BPO9-MgammaG, BPO11-MgammaG, or BPO12-MgammaG, resulted in complete and specific suppression of the induction of the anti-BPO reaginic antibody response without affecting, however, the level of reaginic antibodies to OA. Further study of the effect of epitope density on the immunologic properties of BPOX-MgammaG revealed that a) the lightly haptenated conjugated, BPO1-MgammaG and BPO2.9-MgammaG, were not immunosuppressive, b) the conjugates, BPO4.3-MgammaG and BPO19-MgammaG, were partially tolerogenic, and c) the heavily haptenated conjugate, BPO31-MgammaG, was nontolerogenic. Moreover, most importantly, the ongoing anti-BPO response in sensitized mice was readily abrogated by either four daily or four weekly injections of BPO9-MgammaG. The immunosuppressive effect of BPO12-MgammaG conjugates was dose dependent, complete suppression being achieved with 200 mug of the tolerogen. The unresponsiveness to BPO of spleen cells from immunosuppressed donors was also maintained in adoptive cell transfer experiments in spite of the additional administration of the immunizing antigen under conditions expected to yield a secondary IgE response. Hence, it is suggested that, with special precautions to prevent unleashing an anaphylactic shock, treatment of penicillin-sensitive individuals with polyvalent conjugates of an appropriate number of BPO groups per human gamma-globulin molecule would constitute a rational immunotherapeutic procedure for the abrogation of the allergic response to BPO.


Development of monoclonal IgA and an apparent IgG in a patient with macroglobulinemia: sharing of individually specific antigenic determinants among IgM, IgA, and IgG. Patient CM, who initially was diagnosed as having macroglobulinemia (IgM, kappa) was subsequently found to develop a monoclonal IgA(kappa) protein. Rabbit antisera directed against the patient's IgAm and IgM were rendered specific for individual antigenic (ind) determinants. The anti-IgAm and IgM ind sera reacted with both 131I labeled monoclonal proteins in a double antibody radioimmunoassay (RIA). In addition, both monoclonal immunoglobulins inhibited the reaction between labeled immunoglobulin and both ind antisera, and statistical analysis of the data suggested that the shared ind determinants were identical. The IgG fraction of patient CM's serum also contained a component which competed with both monoclonal IgA (CM) and IgM (CM) in the RIA specific for ind determinants. Analysis of serum samples taken over a 2-year period revealed that, in addition to IgM, both the IgA and IgG components possessing the shared ind determinant(s) were present in low concentrations in the earliest sample, although not detected by conventional techniques. The monoclonal IgA and the IgG component were found to increase in concentration over this time interval with a concomitant decrease in IgM. The regulation of immunoglobulin expression with respect to the proposed models of gene organization in antibody-producing cells was discussed.


Studies of idiotypic antibodies: reactions of isologous and autologous anti-idiotypic antibodies with the same antibody preparations. Outbred rabbits were immunized with keyhole limpet hemocyanin diazotized to p-aminophenyl-N-trimethylammonium chloride (TMA). Anti-TMA antibodies were specifically purified and the F(ab')2 fragments were isolated after pepsin digestion. Glutaraldehyde-polymerized anti-TMA F(ab')2 fragments were reinjected back into the same animal that synthesized the antibodies to elicit an autologous anti-idiotypic (AAI) antiserum and were also injected into an allotypically matched recipient to elicit isologous anti-idiotypic (IAI) antiserum. AAI and IAI antisera reactive with the same F(ab')2 preparation were compared by assaying the populations of anti-TMA molecules reactive with each antiserum. The assays showed that the AAI antiserum reacted with smaller populations of molecules than did the IAI antiserum. The population of molecules recognized by the AAI antiserum was contained within the larger population recognized by the IAI antiserum. Hapten inhibition studies showed that the differences in the populations of molecules recognized by each antiserum were not related to the hapten-binding properties of the idiotypic antibodies: in one case both AAI and IAI reactions were equally inhibited by hapten; in another, the AAI reaction was inhibited more than was the IAI; in another, the IAI reaction was inhibited more.


Autoradiography and immunohistoperoxidase techniques applied to the same tissue section. A method is described which allows simultaneous detection of two different compounds in tissue sections by combining autoradiography and immunohistoperoxidase techniques.


Studies on platelet plasma membranes. I. Characterization of surface proteins of human platelets labeled with diazotized (125i)-diiodosulfanilic acid. A polar, nonpenetrating compound of high specific activity, diazotized (125I)-diiodosulfanilic acid (DD125ISA), has been developed as a label for exposed proteins of the human platelet plasma membrane, and platelet proteins and the pattern of labeling have been studied with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). That DD125ISA binds specifically to membrane proteins was demonstrated by: (1) the specific activity of isolated membrane protein was five to seven times that of whole platelet protein and (2) no proteins of intact platelets were labeled which were not represented in the isolated plasma membrane. That the DD125ISA-labeled membrane proteins were exposed on the cell surface was demonstrated by: (1) DD125ISA-labeled proteins were altered by trypsin treatment of intact, labeled platelets and (2) the pattern of labeling produced by reaction of isolated membranes with DD125ISA was quite different from that produced by the labeling of intact platelets. Analysis of platelet membrane proteins by SDS-PAGE demonstrated the glycoproteins previously described at 150,000 daltons (termed glycoprotein I) and 92,000 daltons (glycoprotein III) but we could discriminate two apparently distinct glycoproteins in the intermediate region (IIa: 125,000 daltons, and II: 118,000 daltons). Glycoproteins I and III were constant whereas IIa was clearly visible only in unreduced samples and II was predominant in reduced samples. Reaction of DD125ISA with intact platelets resulted in equal labeling of three of these four membrane glycoproteins (IIa, II, and III). The pattern of exposed proteins on the platelet surface labeled by DD125ISA was different from lactoperoxidase-131I, which labeled predominantly the 92,000 dalton glycoprotein, as demonstrated by simultaneous SDS-PAGE analysis. Therefore three glycoproteins of the human platelet plasma membrane are exposed to a radioisotope probe on the platelet surface and are accessible for contact interactions.


2,7-Fluorenediamine and 2,5-fluorenediamine as peroxidase reagents for blood smears. Our new histochemical methods for peroxidase activity, which do not require benzidine as a hydrogen donor, have been modified for hematological use. Diamine derivatives of fluorene (2,7-fluorenediamine and 2,5-fluorenediamine) were employed in place of benzidine and the result obtained was satisfactory. Two staining techniques were developed. (1) Smears of peripheral blood or bone marrow aspirates were fixed in 2.5 per cent glutaraldehyde solution for 1 minute. Smears were then washed in tap water and covered either by a saturated solution of 2,7-bluorenediamine or by a 0.05 per cent solution of 2,5-fluorenediamine in the presence of hydrogen peroxide for 5 minutes. After being washed in running tap water they were counterstained with Carazzi's hematoxylin solution for 10 minutes. (2) The second technique employs pretreatment of blood smears with 5 per cent CuSO4 solution and aldehyde fixation was omitted. Smears were counterstained by Giemsa stain. The nuclear chromatin structures were well preserved and nucleoli were easily distinguished in the immature cells. In the first method, clear brown granules were recognized in the cytoplasms. In the second method, peroxidase-positive granules were stained black and the staining of nucleus and cytoplasm resembled that of McJunkin's method.


Cancer of the larynx in Hull and the surrounding area of East Yorkshire. A study of all patients with cancer of the larynx in a population of 449,430 over ten years shows that supraglottic and epilaryngeal cancer has an almost equal incidence to glottic cancer. There is a relationship between urbanization and the incidence of cancer of the larynx, and between cancer of the larynx and bronchogenic carcinoma. A policy of radiotherapy with surgery held in reserve for supraglottic and epilaryngeal cancer failed at a general hospital, for reasons that have been discussed. Primary surgery is required for most cases of supraglottic and epilaryngeal cancer. Radiotherapy with surgery held in reserve is the most appropriate treatment for the great majority of glottic cancers. When radiotherapy failed to cure the disease palliation was achieved in only a small minority of the cases.


Secretory otitis media in chronic lymphatic leukaemia. A temporal bone report of a case treated for seven months with tympanostomy tubes. Serially sectioned temporal bones were studied from a patient with chronic lymphatic leukaemia with submucosal infiltration of the nasopharynx, the resulting secretory otitis having been treated with tympanostomy tubes for 7 months. The initially serous sterile secretion turned mucoid, and a low grade infection supervened. For the last 3 months the tympanostomy tubes were open and the ears dry. In the sections, the mastoid air cells showed leukaemic infiltrations and contained AB- and PAS-positive secretion. The drum openings were without reaction and there was no epithelial ingrowth around the margins. The posterior tympanum in both ears contained organizing fibrous tissue extending from the inner surface of the drum to the promontory. In the tympanum the mucosa was flat or columnar and without glands. Distinct AB and PAS-positive mucus film and secretory cells appeared on the promontory epithelium and the inner surface of the drum. In the tubo-tympanic recess some AB- and PAS-positive subepithelial glands appeared and in the tympanic orifice of the tube the secretory cells were prominent and the lumen full of AB-positive mucus. The case reported represents an intermediate stage in the healing of secretory otitis media, most of the glands having disappeared but the intraepithelial secretory elements being still active.


Cellular events in the induction of experimental allergic encephalomyelitis in rats. Although both the T and B cells of the Lewis rat have immunoglobulin receptors for basic protein (BP) of myelin, and both cell types are required for antibody production to BP, the present results demonstrate that the T cells are the only cells required for the induction of experimental allergic encephalomyelitis (EAE). Both EAE and anti-BP were readily induced in thymectomized, irradiated Lewis rats reconstituted with normal thymus and bone marrow cells and challenged with BP in complete Freund's adjuvant. If the thymus cells were first treated with BP heavily labeled with 125I so as to eliminate (sucide) specific T cells, the recipients neither develop EAE nor produce antibody to BP. On the other hand, if the thymus cells were untreated and the specific B cells of bone marrow were eliminated by treatment with 125I-BP, EAE was not inhibited, although no antibody was produced. These results strongly suggest that the T cell is responsible for the induction of EAE although both the T and B cells are competent to respond to BP. Evidence was presented which suggests that neither suppressor T cells nor circulating antibody are involved in the inhibition of EAE by injection of Lewis rats with nonencephalitogenic preparations of BP. The immune status of T and B cells of the Lewis rat to BP was compared with the immune status of these cells in other species to thyroglobulin, where only the B cells appear to be competent. In this context, Brown Norway rats, which are resistant to the induction of EAE, also appear to lack T cells reactive to BP, although competent B cells are present.


Status of blood group carbohydrate chains in ontogenesis and in oncogenesis. Blood group ABH determinants in human erythrocytes are carried by four kinds of glycolipid carbohydrate chains, differing in their structural complexity. They are Aa, Ab, Ac, and Ad for A variants, and H1, H2, H3, and H4 for H variants (Table I and Fig 1). Based on the surface labeling of A variants and on the reactivity of erythrocytes to antibodies directed against H3 and against its degradation products, it is concluded that complex variants of A or H determinants (Ac and Ad/or H3 and H4) are absent or significantly low in fetal erythrocytes (80-150 days after gestation) and in new born erythrocytes, whereas these complex structures are fully developed in adult erythrocytes. In contrast, A determinants linked to simpler carbohydrate chains (Aa, Ab variants) are fully developed before birth and do not show significant change after birth. The precursor of blood group carbohydrate chains seems to be abundant in fetal or newborn erythrocytes. This assumption is based on the higher reactivity of fetal or newborn erythrocytes to an antibody, which is directed against the precursor N-acetylglucosaminly beta1 leads to 3 galactosyl beta1 leads to 4 glucosylceramide than in adult erythorocytes. Reactions of glycolipids of gastrointestinal mucosa, with antibodies directed against H3 glycolipid and its degradation products, were compared to that of gastrointestinal tumors. The reaction to bela Glc NAc1 leads to 3 beta Gall leads to 4 Glc leads to ceramide (structure 4), which is the precursor of all blood group glycolipids, was consistently high in many cases of tumor glycolipid than that of normal glycolipid. This as well as other evidence supports a general concept that the process of ontogenesis of a blood group carbohydrate chain occurs as step-by-step elongation and arborization, and that blocking of such a development of a carbohydrate chain occurs in the process of oncogenesis.


The surface nature of proteins of a bovine enterovirus, before and after neutralization. The surface nature of the proteins of a bovine enterovirus have been determined by using 125I and pyridoxal phosphate-sodium borohydride labelling techniques. As found previously, 125I labels only VP1 in intact capsid particles, whereas reaction with pyridoxal phosphate followed by reduction with tritiated sodium borohydride labels VP1, VP2 and VP3. Only VP4 is found to have no surface tyrosine, histidine or lysine available for reaction. After neutralization with homologous antisera, however, VP4 becomes exposed and is then available for labelling with 125I. This must reflect a substantial conformational change in the virus particle after neutralization.


Temperature-dependence of rapid axonal transport in sympathetic nerves of the rabbit. Stop-flow techniques were used to determine how temperature affected the axonal transport of dopamine-beta-hydroxylase (DBH) activity in rabbit sciatic nerves in vitro. These nerves were cooled locally to 2 degrees C for 1.5 hr, which caused a sharp peak of DBH activity to accumulate above the cooled region. Accumulated DBH was then allowed to resume migration at various temperatures. From direct measurements of the rate of migration, we found that the axonal transport velocity of DBH was a simple exponential function of temperature between 13 degrees C and 42 degrees C. Over this range of temperatures, the results were well described by the equation: V=0.546(1.09)T, where V is velocity in mm/hr, and T is temperature in degrees centigrade. The Q10 between 13 degrees and 42 degrees C was 2.33, and an Arrhenius plot of the natural logarithm of velocity versus the reciprocal of absolute temperature yielded an apparent activation energy of 14.8 kcal. Transport virtually halted when temperature was raised to 47 degrees C, although only about half of the DBH activity disappeared during incubation at this temperature. Another transition occurred at 13 degrees C; below this temperature, velocity fell precipitously. This was not an artifact peculiar to the stop-flow system since the rate of accumulation of DBH activity proximal to a cold-block also decreased abruptly when the temperature above the block was reduced below 13 degrees C.


Fast axonal transport in motor nerve regeneration. This report describes the fast transport of [3H]-leucine-labeled proteins in regenerating rat sciatic motor nerves. A normal rate of fast transport (383 +/- 33 mm/day) was present in the regenerating sprouts, as well as in the central stumps. The rapidly transported proteins passed the level of axotomy without impediment, and accumulated in the endings of the regenerating sprouts, as shown by electron microscope autoradiography. In addition, transported proteins accumulated in terminal neuromas. The relative amount of protein-incorporated radioactivity in the crest of transport in the regenerating nerves was increased compared to control nerves. These results are interpreted to suggest that the mechanism of fast transport is the same in regenerating nerves was increased compared to control nerves. These results are interpreted to suggest that the mechanism of fast transport is the same in regenerating sprouts as in normal axons; during regeneration fast transport appears to add newly synthesized materials to the growing tip.


Acid maltase deficiency in non-identical adult twins. A morphological and biochemical study. Acid maltase deficiency is described in non-identical adult twins. The onset of the disease can be traced into late infancy; the clinical picture is one of severe muscular dystrophy; respiratory insuficiency was the cause of death in one case. The autopsy showed the central nervous system, heart and liver to be spared. Glycogen filled vacuoles are found in skin, mesenchymal cells, small nerves and skeletal muscles. The light microscopic study of 9 different muscles showed extremely variable involvement ranging from normal appearance to overt vacuolization. A 6--20% residual acid alpha-glucosidase activity was found in visceral organs, cultured fibroblasts and in some skeletal muscles. No satisfactory explanation can be given why this generalized acid alpha-glucosidase deficiency produces a selective involvement of skeletal muscles. If compared with infantile AMD (Pompe's disease) our cases have a much higher residual acid alpha-glucosidase activity and show the presence of an antigenically detectable protein. From our study and from a similar report in the literature (de Barsy et al., 1975), it appears that a combined approach of light microscopy, electron microscopy and biochemical analysis (determination of acid alpha-glucosidase) is necessary to make a diagnosis of AMD in adults.


Lipid composition of myelin in multiple sclerosis. Myelin was isolated from histologically normal white matter and plaques from MS patients and from white matter of neurologically normal controls. No difference was found in the total lipid content. There were no detectable deficits in MS myelin of phosphoglycerides, plasmalogens or sphingolipids. Gangliosides and lysolecithin were not detected. Analysis of the fatty aldehyde composition of the phosphoglycerides and the fatty acid composition of the cholesteryl esters, phosphoglycerides and sphingolipids did not show any differences between the normal and MS myelin.


[Neurological findings (especially "developmental reflexes") in residents of homes for the aged (author's transl)]. 86 residents of several homes for the aged in Berlin, aged 75 to 96 years (mean age 86.3 years), were examined neurologically. Many showed the following aberrant neurological signs: Pallaesthesia and dermolexia were extinct in the lower extremities; the ankle jerks could not be elicited; the palmomental, orbicularis oris reflex, grasping and the snout reflexes were positive; there was a hypokinetic-hypertonic motor syndrome. The statistical analyses and the follow-up dynamics of our findings argue for such age-dependent changes of the CNS as the basis for the grasping and snout reflexes and for the involuntary movements of the elderly.


Some chemical principles applicable to some silver and gold staining methods for neuropathological studies. Some of the chemical principles involved in some silver and gold staining procedures for neuropathological studies are analyzed in the light of experience with methods for astrocytes, microglia, axons and reticulin fibers. The role of formalin fixation, and of treatment with ammonium bromide or oxidizing agents prior to staining for glial cells or reticulin fibers respectively, is described. The silver staining methods are divided into two fundamentally different groups. One is based on the use of silver diammine solutions with or without other substances such as carbonates or pyridine, and at varying times, gold toning. This group encompasses a great variety of techniques for staining glial cells, reticulin fibers and axons. The second group is based on the use of weak solutions of silver ions, reduction in a photographic developer, treatment with gold chloride, and a second reduction in oxalic acid. This group is represented by methods for staining axons and reticulin fibers. A brief analysis of the gold and mercuric chloride methods for astrocytes is also given.


Optic neuritis in relation to multiple sclerosis. Available estimates of the frequency with which a patient with optic neuritis develops multiple sclerosis range from as low as 13% to as high as 87%. In an effort to obtain a better estimate, a nation-wide study of optic neuritis was carried out in Israel. Patients who fulfilled strict diagnostic criteria of optic neuritis were identified and examined periodically. Between 1955 and 1964, 105 patients were found and on the basis of these, the average annual age-adjusted incidence of optic neuritis in Israel was 0.56 per 10(5) population compared to 1.2 per 10(5) cases of multiple sclerosis per year, i.e. optic neuritis was about half as frequent as multiple sclerosis each year. As with multiple sclerosis, optic neuritis was more common in European immigrants to Israel than Afro-Asian immigrants. During a follow-up interval which ranged from 3.3 to 15.6 years (mean 9.5 years), at least 27 of the 105 patients developed multiple sclerosis (28%). A life-table analysis showed that after 10 years 32.3 +/- 5.6% of patients with optic neuritis would develop multiple sclerosis and, after 14 years, about half would develop multiple sclerosis. Risk of dissemination was highest in those who were youngest when optic neuritis developed. Neither sex nor ethnic background influenced risk significantly. Results of the present study support earlier work using life-table methods carried out in Hawaii which also showed that between 29 and 39% of patients with optic neuritis will develop multiple sclerosis within 10 years of onset. The life-table method is a better predictor of prognosis than newer laboratory techniques such as spinal fluid studies of IgG, kappa-lambda light chain ratios and serum/CSF IgG ratios.


[Methods and evaluation of visually evoked EEG potentials in cases of suspected multiple sclerosis (author's transl)]. In multiple sclerosis, average EEG potentials which are monocularly evoked by checkerboard pattern reversal frequently show increased latencies of the dominant, occipitally positive peak (more than 110 ms), and latency differences of more than 6--7 ms between responses to right and left eye stimulation. Fixation of the stimulus field at the lower border causes significantly longer latencies and smaller amplitudes than fixation at the upper border. With lower border fixation, the increase of response latency may suggest a reversal of response polarity in extreme cases. Central fixation often but not always results in responses similar to upper border fixation. In order to have minimal variability of the results, fixation of the stimulus field at the upper border is preferred over central fixation.


Effects of modification of intracellularcyclic AMP levels on the imediate hypersensitivity reaction in vivo. Experiments were performed to study the effects of agents that elevate endogenous cyclic AMP levels on the progression of a reverse passive Arthus reaction in the pleural cavity of rats. Three hours after the onset of the reaction a marked reduction in histamine release was observed which correlated well with increases in cellular cyclic AMP concentration. The results at 6 hr suggested a relationship between cyclic AMP and leucocyte PGE, the latter being reduced dramatically both in the cells and in the fluid exudate by agents that elevate cyclic AMP. It was concluded that cyclic AMP may play an important regulatory role in vivo in the release of inflammatory mediators from leucocytes.


The endometrial identity of benign stromatosis of the ovary and its relation to other forms of endometriosis. A series of 114 ovaries form women aged 20 to 82 showed cortical islands of endometrial stroma in about two-thirds of those over 40. This identity was commonly apparent in ordinary sections and was confirmed in half the affected ovaries by the presence of K cells (endometrial granulocytes) in one or more foci; it may be masked by attenuation or conversion into smooth muscle. The islands show transitional forms to decidual foci and to cortical granulomata and related leucocytic lesions, a few of which also showed K cells: they are ultimately converted into weakly orceinophil hyaline foci which are common in later life and showed K cells in four instances and continuity with stromal islands in 20 ovaries. The lesions appear to arise from the implantation of endometrial stroma into the ovary, the surface of which becomes more cellular and probably more permeable after 40. The islands may foster the development of cortical stromal hyperplasia and their antecedents that of surface epithelial tumours.


Alpha fetoprotein in the antenatal diagnosis of the congenital nephrotic syndrome. In the congenital nephrotic syndrome (Finnish type), concentrations of alpha fetoprotein in the amniotic fluid and maternal serum are markedly elevated in the second trimester of pregancncy. Demonstration of this alteration allows early prenatal diagnosis of this fatal condition and elective termination of the pregnancy before 20 week's gestation.


Indications for the use of bleomycine-Co57 in brain scanning. The Authors report on their experience with a new radio-active tracer, Bleomycine-Co57, which was studied in 40 patients who had previously undergone brain scanning with Indium113m. The results obtained with these two different tracers are compared and the following conclusions advanced: 1) Bleomycine-Co57 is the radio-active tracer of choice in identification of cerebral metastases, especially in those cases where brain scanning with Indium113m was non-diagnostic; 2) glioblastomas evidence less uptake of the tracer than do metastases; 3) the scarce affinity of this tracer for benign tumors is stressed; 4) owing to the long half-life of this tracer, it is preferable to use it only with those subjects suspected of having a malignant neoplasia.


Vinyl acetate casts of emphysematous rat lungs. Emphysema was induced in rats by chronic exposure to a papain aerosol according to established methods. The emphysematous changes were evaluated by previously described histological and biochemical parameters and by a new method employing infiltration of the lungs with vinyl acetate to make casts of the air spaces of the pulmonary tree. The weights and gross and microscopic appearance of the casts demonstrated the emphysematous changes in lungs of the papain-treated rats. Vinyl acetate casting may offer a new and useful tool in evaluating experimental induction of emphysema in rats.


Alterations in amounts and rates of serotonin transported in an axon of the giant cerebral neurone of Aplysia californica. 1. The giant cerebral neurone of the sea hare, Aplysia californica, is a unipolar serotonergic cell. Its axon bifurcates, one branch travelling in the cerebrobuccal connective, the other in the posterior lip nerve. 2. I13H]serotonin was injected under pressure into the cell body of the giant cerebral neurone. We studied fast axonal transport of the radioactive transmitter substance along the lip nerve when the cerebrobuccal connective was cut close to the bifurcation. 3. When the connective was cut, more than twice as much [3H]serotonin was transported along the lip nerve compared to uncut control nervous systems. 4. The increased [3H]serotonin appearing in the nerve probably was originally destined to enter the connective, but was diverted from the cut stump which was occluded with backed-up material. 5. The incremental [3H]serotonin in the lip nerve was not the result of increased export from the soma in response to injury. 6. Not only was more [3H]serotonin transported along the lip nerve, but also a far greater fraction of the transmitter moved at very fast transport rates, approaching 120 mm/day. In uncut control nerves only a small fraction of total [3H]serotin moved faster than 70 mm/day. 7. These results are interpreted with a model for fast axonal transport. We suggest that serotonergic vesicles move at a fixed, maximal speed when attached to essentially immobile tracks, but that the vesicles are only intermittently associated with the tracks. We presume that the rate-limiting step in movement of vesicles is the concentration-dependent and reversible binding to the tracks. Transport along axons may be considered analogous to those enzymatic reactions in which formation of the enzyme-substrate complex limits the appearance of product. Translocation is here analogous to formation of product. The process may therefore be approached theoretically by modification of the Michaelis-Menten formulation.


An electromyographic study of uterotubal activity in the ewe. The electrical activity of the oviduct and uterus was automatically registered on a potentiometric pen recorder in ewes fitted with an extracellular multielectrode assembly. The duration and amplitude of local non-propagating activity of the uterus increased and became progressively grouped into phases as natural oestrus progressed. Phases of contractions lasting 5-6 min were initially propagated at a rate of 2/hr. Their frequency increased within 24 hr of the decline in plasma progesterone levels. Grouped activity was then resumed for the next 24 hr. Similar changes were seen in the activity of the oviduct although this region was active earlier in oestrus. The uterotubal activity developed in the same way 24 hr after withdrawal of a progestagen-impregnated sponge but lasted for 3 days. When another oestrus was induced by injection of a prostaglandin analogue, the activity patterns were qualitatively similar although they did not start until 36 hr after injection and lasted for only 2 days.


Type C viruses from Kirsten sarcoma-transformed mink cells co-cultivated with primate cells and expressing p30 antigens related to feline leukemia virus. Two type C viruses with new antigenic and biological properties were isolated by co-cultivating secondary cell strains established from the kidneys of a baboon (Papio papio) and a patas monkey (Erythrocebus patas) with mink cells non-productively transformed by Kirsten sarcoma virus. Both new isolates (designated PP-1R and EP-1R) contain major structural proteins (p30) that are immunologically most closely related to the p30 proteins of feline leukemia viruses. The reverse transcriptases of both viruses, although antigenically related to polymerases of murine and rat type C viruses, are distinct from those of previously described type C viral groups. Both PP-1R and EP-1R can be transmitted to canine and feline cells and to sarcoma virus-transformed, but not normal, mink cells. Both viruses contain RNA genomes partially homologous to those of endogenous mouse and rat type C viruses and the Kirsten sarcoma virus. In addition, the RNA of PP-1R contains a portion of the nucleic acid sequences found in a type C virus isolated from the baboon species P. papio. We propose that both new isolates are genetic recombinants formed between endogenous primate type C viral genomes and sequences found in Kirsten sarcoma-transformed mink cells.


Recombination between a temperature-sensitive mutant and a deletion mutant of Rous sarcoma virus. Cells doubly infected with two mutants of the Schmidt-Ruppin strain of Rous sarcoma virus (RSV), ts68, which is temperature sensitive for cell transformation (srcts), and a deletion mutant, N8, which is deficient in the envelope glycoprotein (env-), produced a recombinant which carried the defects of both parents. The frequency of formation of such a recombinant was exceptionally high and made up 45 to 55% of the progeny carrying the srcts marker. By contrast, the reciprocal recombinant, which is wild type in transformation (srcts) and contains the subgroup A envelope glycoprotein (envA), was almost undetectable. This remarkable difference in the frequency of the formation of the two possible recombinants suggests that a unique mechanism may be involved in the genetic interaction of the two virus genomes, one of which has a large deletion. When an RNA-dependent DNA polymerase-negative variant of the N8 (N8alpha) was crinants also became deficient in the polymerase. Cells infected by the srctsenv- recombinant were morphologically normal at the nonpermissive temperature (41 degrees C) and susceptible to all subgroups of RSV. The rate by which the wild-type RSV transformed the recombinant-preinfected cells was indistinguishable from that of transformation of uninfected chicken cells by the same wild-type virus. This indicates that no detectable interference exists at postpenetration stages between the preinfected and superinfecting virus genomes and confirms that the expression of the transformed state is dominant over the suppressed state.


Case report: potassium of 10 milligrams per cent. An 82-year-old white man with recurrent prostatitis was hospitalized with acute urinary retention. On 3 separate occasions the blood potassium was 10 mg. per cent. An admission electrocardiogram was typical of hyperpotassemia. The patient recovered on intravenous therapy and conservative treatment. Transurethral resection of the prostate was done 3 weeks later.


Prenatal diagnosis of neural tube defects. A prospective program for the prenatal detection of neural tube defects in a high-risk pregnancy group has been under way for one year. Twenty high-risk mothers have undergone a complete prenatal work-up, including sonography, amniocentesis, and amniotic fluid and maternal serum alpha-fetoprotein quantitation. Within this group, two cases were prospectively diagnosed and were later confirmed.


[Effect of continuous bleomycin treatment and of oil-suspended bleomycin on experimental tumor growth (author's transl)]. Effects of continuous administration of bleomycin solution and of intralesional injection of sesame oil-suspended bleomycin on tumor growth were studied. Experimental animal tumors were 3 rd generation isotransplants of a spontaneous C3H mouse mammary carcinoma. Bleomycin treatments were started when transplanted tumors reached 8 mm in diameter and the measurement of tumor volume was followed. Dose administered was fixed as 100 mg/kg in all the groups. Bleomycin solution was given intralesionally in a single or 4 daily doses, or intraperitoneally by continuous infusion. The latter method inhibited tumor growth most effectively, while the single injection was the last effective. Intralesional injection of oil-suspended exhibited similar effectiveness as the continuous infusion, and it was independent of the number of fractions. These results were interpreted by the several features in the response of mammalian cells to the antibiotic.


Lung damage caused by phospholipase C and the changes in phospholipids in the rat lung. The lipid and protein fractions of the endobronchial lavage fluid from the normal rats which contained the lung surfactant were analysed. Lecithin, the main main component of the lung surfactant, was exclusively combined with a dextran precipitable protein. This protein then behaved as beta-globulin on cellulose acetate electrophoresis or low density lipoprotein on agarose gel filtration. After administration of phospholipase C (from Clostridium Welchii), the protein content of the lavage fluid increased markedly. The amount of dextran precipitable protein also increased markedly and its properties remained the same on gel filtration after treatment with phospholipase C. The phospholipids in the lavage fluid were not affected, although the total phospholipids in the lung tissue, especially lecithin, did decrease during the 10 days after treatment. Histopathologically, an eosinophilic dense exudative fluid appeared in both the interstitium and the broncho-alveolar spaces. A large number of the alveolar lining cells disappeared and a few of them were desquamated into the alveolar spaces. Thus, the immediate destruction of the alveolar lining cells after the administration of phospholipase C resulted in interstitial pneumonia in 10 days. The significance of phospholipase in pulmonary inflammation is discussed.


Pseudomonas aeruginosa antibodies in human plasma. Plasma samples from healthy adults were examined for antibodies to the common protective antigen (OEP) [1] by passive hemagglutination (HA) reaction [2] with the finding that the majority of them had an antibody titer of 60 or lower. 2-Mercaptoethanol (2-ME) treatment, however, caused a decrease in HA titer down to 16 or lower in most of the cases, suggesting that the OEP antibody resides mostly in IgM and slightly in IgG. The finding was corroborated by gel filtration on Sephadex G-200. It appears likely that IgA is not implicated in the HA titer since IgA was HA negative. However, the detection of enzymatic activity by enzyme-linked immunosorbent assay (ELISA) [9] still suggests a possibility that OEP antibody resides in IgA. On the other hand, antibodies to proteolytic enzymes, especially protease and elastase [3], elaborated by Pseudomonas aeruginosa, were found to be present in most cases at a titer of 16 or lower by HA reaction. It was also found that they reside mostly in IgM.


Antiglobulin factor in sera from patients with liver disease. (1) An antiglobulin factor, non-neutralizable by human gamma-globulin, was demonstrated in sera of two patients with liver disease. (2) By absorption and elution techniques, two fractions were differentiated in a serum: one is reactive to rabbit antibody and the other seems cross-reactive to rabbit and human antibodies. (3) In double immunodiffusion test, the antiglobulin factor formed a precipitation band with heat-aggregated human IgG as did rheumatoid arthritis serum. (4) While the antiglobulin activity to rabbit antibody was demonstrated in both the IgM and IgG fractions, the reactivity to human antibody was localized in the IgM fraction. (5) From its selective reactivity to individual anti-D sera (sensitizers), at least a part of the specificity of the antiglobulin factor must be related to anti-Gm (1), and consequently can be regarded as auto-reactive.


Whipple's disease: demonstration by immunofluorescence of similar bacterial antigens in macrophages from three cases. Although it is widely accepted that Whipple's disease is caused by microorganisms, there is little agreement as to the exact nature of those microbes. Several different types of bacteria have been reported from patients with Whipple's disease causing some workers to speculate that Whipple's disease may be due to a variety of microorganisms. Using an indirect immunofluorescence technique, we have demonstrated bacterial antigens in the granules of the foamy macrophages from three patients with Whipple's disease. The macrophages in each case stained with several types of antibacterial grouping sera. Jejunal biopsies from three normal subjects and one from a patient with celiac-like disease did not show significant reactions with any of the antisera. When these same antisera were reacted with homologous and heterologous bacteria, numerous cross-reactions were seen. However, each organism only reacted with certain antisera resulting in a distinctive pattern of reaction which could identify it. Therefore, if a different organism was present in each case of Whipple's disease, the antisera should give dissimilar reactions from case to case, but if the same type of microbe was present in each case, a similar pattern of reaction should be seen. In fact, the reactions were remarkably similar from case to case. These results suggest that a single, antigenically definable microorganism is present in the jejunal tissue of patients with Whipple's disease.


Appearance, properties, and origin of altered human hemoglobin in feces. Altered hemoglobin (Hb) has been found in the feces as a sequel to an upper gastrointestinal bleed. Active Hb antigen of increased anodic mobility was detected on immunoelectrophoresis of melena stools using a goat anti-Hb. The Hb derivative was also identified in polyacrylamide gel electrophoresis using 412 nm. absorbance. The alteration could be simulated in vitro by incubation of hemolysate with duodenal juice or purified carboxypeptidase B alone, or by a mixture of carboxypeptidases A and B. Treatment of hemolysate or purified Hb with acid, gastric juice, pepsin, pancreatic juice, bile, trypsin, or chymotrypsin failed to produce the characteristic alteration. Instead, no change, or production of alpha and beta chains, or gradual but complete elimination of the Hb antigen was seen. This latter all or none pattern is presumed to prevail in the large bowel on the basis of incubations of hemoglobin-feces mixtures. Individuals documented to be bleeding into the colon were found to have at least a portion of their Hb antigen in the unaltered form by immunoelectrophoresis. This finding may be of value in identifying the general origin of a gastrointestinal bleed.


Administration of human fibroblast interferon in chronic hepatitis-B infection. One patient with hepatitis-B surface antigen (HBsAg)-positive chronic aggressive hepatitis, and two chimpanzee carriers of HBsAg, were each given seven doses of 10(7) I.U. of human fibroblast interferon over two weeks. The main differnce observed after treatment was a depression of the nucleocapsid hepatitis-0 core antigen in the liver, indicating that hepatitis-B virus infection is sensitive to interferon. Except for a short febrile reaction, no undesirable effects were seen after the administration of human fibroblast interferon which has not been previously given to man.


Identification of high-risk patients with aplastic anaemia in selection for allogeneic 0one-marrow transplantation. Of 75 patients with asplastic anaemia treated between 1968 and 1975, 33 were retrospectively considered as potential candidates for allogenegic bone-marrow transplantation on the basis of their age and severity of marrow failure. The prognosis  of these patients with conservative treatment was assessed from parameters obtained at the time of the initial diagnosis. Initial peripheral-blood granulocyte or platelet concentrations were not of porgnostic value. In contrast, initial reticulocyte concentrations, allowed separation of the patients into two groups with poor and good prognosis. Low initial reticulocyte concentrations (less than 10 000/mu1) indicated those patients at extremely high risk of succumbing to their marrow aplasia (there were no survivors 36 months after disgnosis). In contrast, 75% of those patients with more than 10 000 reticulocytes per mu1 at diagnosis survived for 3 years. Initial peripheral-blood reticulocyte concentrations thus appear to indicate the extent of the marrow failure in aplastic anaemia more accurately than granulocytes or platelets. Low initial reticulocyte concentrations may indicate, among patients with severe aplastic anaemia, those for whom allogeneic bone-marrow transplantation should be seriously considered; patients with higher initial reticulocyte concentrations may benefit from conservation treatment.


Serum-25-hydroxy-vitamin-D in untreated parenchymal and cholestatic liver disease. Serum-25-hydroxy-vitamin-D (25 OHD) concentration has been measured in 106 patients with untreated parenchymal and cholestatic liver disease. Low mean values were found in groups of patients with alcoholic hepatitis and cirrhosis, non-cirrhotic active chronic hepatitis, lupoid and cryptogenic cirrhosis, symptomatic primary biliary cirrhosis, and acute and chronic biliary disease. In a group of patients with presymptomatic biliary cirrhosis the mean value was not significantly different from normal. It is concluded that in the presence of significant parenchymal or cholestatic liver disease serum-25-OHD concentrations are usually low. The mechanisms for the reduction remain to be clarified, but low serum-25-OHD values may play a contributory role in the aetiology of osteomalacia in chronic liver disease.


HLA and pancreatic islet cell antibodies in diabetes. The frequency of HLA-B8 was significantly increased in pancreatic islet cell antibody (P.I.C.A)-positive patients (61%) compared with P.I.C.A.-negative patients (35%) and a control population (28%). This increased frequency of HLA-B8 was even more striking in diabetics in whom P.I.C.A. persisted for more than 5 years (71%). Thus the association of HLA-B8 with diabetes may be related to the presence of P.I.C.A. in these patients.


Serum-prolatic in women with premenstrual syndrome. Serum-prolactin was measured by radioimmunoassay during the menstrual cycle in 28 women who had the premenstrual syndrome (P.M.S.) and in a control group who did not have P.M.S. symptoms. Throughout the menstrual cycle mean serum-prolactin was signficantly higher in women with P.M.S. than in the controls. The average individual increase in serum-prolactin during the premenstrual period compared with the first 3 weeks of the cycle was also significantly higher in women with P.M.S. It is not clear whether the increase in serum-prolactin was merely an indicator of stress or was actually involved in producing some of the symptoms which constitute P.M.S.


Beta-lactamase-producing, penicillin-resistant gonococcus. A gonococcus was isolated from the vagina of a woman with pelvic inflammatory disease. The organism was typical in all respects except that, unlike any previously described gonococcus, it produced a beta-lactamase that conferred resistance to penicillin, ampicillin, and cephaloridine. Among other antibiotics tested streptomycin was the only one to which the organism was resistant.


Penicillinase-producing Neisseria gonorrhoeae. Repeated failure of penicillin G treatment in a man with gonorrhoeal urethritis prompted penicillin sensitivity testing of the responsible strain of Neisseria gonorrhoeae. Disc diffusion sensitivity testing showed complete resistance to 10 mug of penicillin G. The organism produced penicillinase.


Use of nasal continuous positive airway pressure to treat severe recurrent apnoea in very preterm infants. Nasal continuous positive airway pressure (C.P.A.P.) of 2-3 mm Hg abolished or reduced the incidence of severe apnoeic attacks in 5 very preterm newborn infants. It is postulated that C.P.A.P. provides a respiratory drive by reflexly stimulating the infant's pulmonary stretch receptors.


A model for the persistence or reactivation of fetal haemoglobin production. Normal adults may have two distinct erythroid precursor populations, a major one which produces only adult haemoglobin (HbA), and another which produces HbA and fetal haemoglobin (H0F) (F cells). Persistence or apparent reactivation of HbF production in adults results from differential selection of these F cells, except in those rare conditions which are due to specific deletions of D.N.A. involved in suppression of gamma-chain synthesis. The increase in HbF which results from a genetically determined increase in F cells appears to ameliorate sicke-cell anaemia or beta thalassaemia. Augmentation of the F-cell population might offer a therapeutic approach to these disorders.


Clinical syndromes of mastalgia.

Clinical syndromes of mastalgia. 232 patients attending a breast clinic with breast pain as the primary presenting symptom were studied prospectively to define clinical syndromes and to attempt to elucidate aetiological factors. Those women in whom mastalgia was a minor aspect of their complaint, or who were primarily seeking reassurance that they did not have cancer, were excluded. Most mastalgia patients could be placed into well-defined subgroups on the basis of clinical, radiological, and pathological features. After excluding causes of pain arising outside the breast, six specific groups with widely differing aetiological bases were defined, leaving only 7% unclassified lithout known aetiology. The six defined groups were cyclical pronounced mastalgia, (believed to be hormonally based), duct ectasia. Tietze syndrome, trauma, sclerosing adenosis, and cancer. Psychological factors were found to be less important than has been previously suggested. Classification of patients with mastalgia into homogeneous subgroups is a prerequisite of any therapeutic study.


Aprospective study of urinary-tract infections in a Dutch general practice. In a Dutch general practice the adult female population was screened for asymptomatic bacteriuria (A.B.) by repeated urine culture after vulval cleansing. The prevalence of signficant A.B. was 4.7% and increased with age. Women with significant A.B. were followed up for one year. All symptomatic urinary-tract infections were recorded during the same period (incidence 59 per 1000 population). Women with significant A.B. at screening were divided into three groups according to the pattern of the follow-up results: transitory A.B., symptomatic A.B., and persistent A.B. The last group differed from the penultimate group with regare to the site of their urinary-tract involvement, symptomatic cases having predominantly upper-urinary-tract involvement and persistent cases lower-urinary-tract infection. In the matched control group the acquisition-rate of both symptomatic and asymptomatic 0acteriuria was over 12%, a figure similar to the percentage of women present in the practice population during one year with transient, symptomatic, and persistent A.B. Screening for A.B. in the general non-pregnant female population is not advocated at present. Screening and treatment of existing A.B. should be carried out in pregnant women who run an increased risk.


Use of coagulation tests to predict the clinical progress of pre-eclampsia. The clinical manifestations of severe pre-eclampsia are normally separated from those of mild pre-eclampsia and normal pregnancy on arbitrary grounds. A clinical index, based on the increase in diastolic blood-pressure and the presence of proteinuria, was developed to reflect the spectrum of disease from mild to severe pre-eclampsia. This was related to a coagulation index based on the platelet-count, plasma-factor-VIII, and serum-fibrinolytic-degradation-products. The two indices were shown to be strongly correlated. All cases of perinatal death associated with pre-eclampsia had coagulation indices in the most severely abnormal range. These results suggest that intravascular coagulation is a highly characteristic feature of pre-eclampsia and that the coagulation index may be of value in  monitoring the progress of the disease.


Single-dose peroperative antibiotic prophylaxis in gastrointestinal surgery. A single intravenous dose of tobramycin and lincomycin, given at the start of gastrointestinal operations, significantly reduced the incidence of postoperative wound infection from 34% to 5%. The occurrence of both anaerobic and aerobic bacteria was reduced. Therapeutic concentrations of the antibiotics were maintained throughout the operative period in most cases. No toxic effects of the antibiotics were detected, no anaesthetic complication occurred, and resistant strains of bacteria normally sensitive to the antibiotics were not isolated from wounds.


Effects of timolol and hydrochlorothiazide on blood-pressure and plasma renin activity. Double-blind factorial trial. The effects of timolol (10 mg thrice daily) and hydrochlorothiazide (50 mg/day) have been compared in a double-blind factorial trial in 20 patients with essential hypertension. There were four randomised test phases of 8 weeks each during which patients received timolol alone, hydrochlorothiazide alone, timolol plus hydrochlorothiazide, and no treatment (placebo). Blood-pressure was measured weekly, alternately at the outpatient clinic and at the patient's home. Supine mean arterial pressure fell from 119 mm Hg in the placebo phase to 110 mm Hg in the hydrochlorothiazide phase, 106 mm Hg in the timolol phase, and 101 mm Hg in the combined timolol plus hydrochlorothiazide phase. Factorial analysis revealed that these effects of the two drugs were additive without any potentiation or antagonism. Mean plasma-renin activity (P.R.A.) was 5-02 ng/ml/3 h in the placebo phase falling to 1-79 in the timolol phase and rising to 9-54 in the diuretic phase, but remaining unchanged in the combined treatment phase (5-40 ng/ml/3 h). The data suggest that the hypotensive action of timolol is not dependent on the concomitant fall in P.R.A. The methods described provide a valuable tool for quantitating the effects of a given drug, and hence a valid basis for objective comparison.


Exercise vectorcardiography in diagnosis of ischaemic heart-disease. The changes in the cardiac electric field during exercise have been studied by conventional vectorcardiographic techniques. Criteria have been developed to distinguish the changes seen in patients with coronary-artery disease proven angiographically from the changes in patients with normal coronary arteriograms. These criteria were derived from a learning set of 105 patients. Vectorcardiograms were taken immediately before and immediately after a maximal treadmill exercise test. In this group of 105 patients--72 with abnormal coronary arteries and 33 with normal coronary arteries--vectorcardiography before and after exercise diagnosed correctly 86% of the abnormals and 82% of the normals. This gives a sensitivity of 86% and a specificity 82%. ST-segment analysis in the same group gave a sensitivity of 60% and a specificity of 85%. The same criteria applied blind to a second testing set of 92 patients gave a sensitivity of 84% and a specificity of 65%. In the same group ST-T analysis gave a sensitivity of 56% and a specificity of 78%. The sensitivity with these criteria compares favourably with the best results so far achieved with ST-segment analysis aided by computer. This technique can be easily used by the clinician and improves the diagnostic accuracy of exercise testing.


Clinical features and follow-up of patients with angina and normal coronary arteries. The clinical findings in 45 patients with angina and normal coronary arteries are reviewed. The primary site, radiation, and character of the pain were typical of angina but the pain was atypical in its relation to stress, frequency of occurrence, relief with rest, and response to nitroglycerin. 22 had abnormal electrocardiograms with evidence of past myocardial infarction in 3. 5 had abnormal exercise tests. During a two-year follow up period there were no further myocardial infarctions and anginal pain either disappeared or improved in 73%. It is concluded that patients with angina and normal coronary arteries can often be distinguished clinically and that they have a good prognosis.


Oestrogen receptors in human malignant melanoma. 16 of 35 patients (46%) with malignant melanoma were found to have cytoplasmic oestrogen-receptor activity in biopsy specimens. OEstrogen-receptor activity was detected in primary lesions, lymph-node metastases, skin metastases, and visceral metastases. Equal percentages of males and females had positive assays. Scatchard analysis of binding in one case was consistent with a single class of high affinity receptor sites.


Cotrimoxazole and folate metabolism.

Cotrimoxazole and folate metabolism. Cotrimoxazole 4 tablets daily (1 tablet = trimethoprim 80 mg and sulphamethoxzole 400 mg) was given for a period of six to fourteen days to 13 inpatients, and serum-folate levels were measured before and one day after the course of treatment. The results were compared with those from 8 patients not receiving antibacterial therapy, tested on admission and one week later. Two assay techniques were used, one employing Lactobacillus casei and the other 125I-labelled folate isotope dilution. The microbiological technique showed a significant decline in folic-acid levels in the serum after cotrimoxazole, and this decline was not seen in controls. By contrast, the radioisotope technique showed no significant alteration in serum-folate levels compared with controls. This suggests that cotrimoxazole does not depress true serum-folate and that many low microbiological results obtained during cotrimoxazole therapy reflect interference with the assay organism. There is insufficient evidence to incriminate cotrimoxazole as a significant cause of blood dyscrasias in excess of those which might occur on sulphonamide alone or even with other antibacterials.


Visualisation of thrombi with technetium-99m urokinase. A negative report. Direct imaging of thrombi with isotopically labelled agents would provide a convenient atraumatic method of diagnosing deep venous thrombosis. Urokinase labelled with technectium-99m has many theoretical advantages and successful use of this agent has been reported. A method for tagging urokinase with Tc-99m has been developed which preserves the clot-lysing ability of the urokinase. The thrombus imaging previously reported has not been duplicated.


Effect of low-dose subcutaneous heparin on whole-blood viscosity. The change in blood-viscosity at low shear-rates (0.77 s-1 and 2-62 s-1 was measured in eighteen normal subjects and postoperatively in sixteen patients after administration of 5000 I.U. of subcutaneous heparin. In both groups there was a significant decrease in the mean blood-viscosity 4 to 6 hours after the injection of heparin. This fall in blood-viscosity may be involved in the prophylactic effect of low-dose subcutaneous heparin in preventing venous thrombosis.


Correction of hypocalcaemic symptoms during plasma exchange. The supplementation of replacement solutions used during plasma exchange with calcium gluconate prevents hypocalcaemic symptoms.


Mechanisms by which amniotic-fluid alpha-fetoprotein may be increased in fetal abnormalities. Assuming that urination is the principle route by which alpha-fetoprotein (A.F.P.) passes into the amniotic fluid from the normal fetus, and that A.F.P. is mainly degraded by a process of fetal swallowing and digestion, 4 possible mechanisms by which amniotic-fluid A.F.P. is increased by fetal abnormalities are postulated.(1) Amniotic-fluid A.F.P. is increased in open neural-tube defects by leakage of A.F.P. from fetal serum and cerebrospinal fluid.(2) In exomphalos exposure of blood-vessels in the extruding viscera permits transudation of A.F.P. into the amniotic fluid.(3) In nephrosis, fetal proteinuria increases amniotic-fluid A.F.P.(4) Impaired fetal swallowing or digestion would account for increased amniotic-fluid A.F.P. in congenital malformations of the orogastrointestinal tract.


Cause of analgesic-induced renal papillary necrosis. It is proposed that analgesic-induced papillary necrosis is due to a progressive narrowing of small blood-vessels of the renal papilla and renal pelvis.


Tuberculosis in the Potteries 1971-74.

Tuberculosis in the Potteries 1971-74. Notification rates of all forms of tuberculosis have increased in all age-groups in the Potteries, in a stable population which includes only a small immigrant community. The increase is greatest in both males and females over the age of 65 years and under 15 years. High notification rates have been recorded in workers in the pottery and mining industries and were unrelated to pneumoconiosis. Discrepancies have been found between the numbers of notified cases and the numbers of laboratory isolates of tubercle bacilli, particularly in cases of non-respiratory disease. This indicates that not all proven cases of tuberculosis are notified. The study has revealed serious deficiencies in the contact-tracing procedure in certain areas of the Potteries consequent upon the closure and reorganisation of outlying chest clinics.


Transformation of lymphocytes from patients awaiting cadaver renal transplants. The response of lymphocytes from prospective kidney transplant recipients to inactivated donor lymphocytes (one-way mixed-lymphocyte reaction (M.L.R.)) and to phytohaemagglutinin (P.H.A. response) was measured before transplantation of cadaver kidney allografts in 78 and 75 cases respectively. M.L.R. results from patients whose grafts had failed within 3 or 6 months compared with M.L.R. results from patients whose grafts continued to function at the same times showed only slight differences irrespective of whether the data was expressed as a relative response or a mitotic index. Because of the large overlap in values, it was concluded that the M.L.R. test was of limited value in predicting graft prognosis. Pretransplantation P.H.A. responses from patients whose grafts failed within 3 or 6 months were significantly different from those shown by patients whose grafts remained functioning at those times. It is suggested that lymphocyte responsiveness to P.H.A. before transplantation may be of value in predicting the fate of renal allografts.


Protein and calorie malnutrition, cell-mediated immunity, and B.C.G. vaccination in children from rural West Bengal. After B.C.G. vaccination of a group of tuberculin-negative children in West Bengal, India, 197 have been retested with tuberculin. 35-5% showed a definitely positive reaction. Only 15% showed no response to B.C.G. When compared by weight for age, the rate of positivity of those below 60% of the Harvard standard was significantly lower than of those above 80%. Those between 60 and 80% were similar to those above 80%. However, when the group was subdivided according to protein and calorie nutrition as measured by arm muscle and fat cross-sectional areas, the difference was striking. The tuberculin-test response was grossly impaired in those who were primarily severely protein deficient (kwashiorkor type), significantly depressed in those who were both severely protein and severely calorie deficient (marasmic-washiorkor type), and not depressed in those who were severely calorie deficient but normal or low in protein (marasmic type). It is suggested that clear distinction between different types of nutritional deficiency in a given geographic area is of direct relevance in planning mass vaccination programmes in that area.


Host responses to hepatitis-B infection in patients with primary hepatic carcinoma and their families. A case/control study in Senegal, West Africa. A case/control study of patients with primary hepatic carcinoma (P.H.C.) and their families was carried out in Dakar, Senegal. 28 P.H.C. cases were matched by age,sex, and ethnic group with 28 controls. Serum was collected from cases, controls, parents (28 mothers, 27 fathers) of cases, parents of controls, 71 siblings of cases, and 58 siblings of controls. Assays of their sera for hepatitis-B surface antigen (HBsAg), antibody to HBsAg (anti-HBs) and antibody to hepatitis-B core antigen (anti-HBc) produced the following results. (1) Nearly all P.H.C. cases (97%) and controls (93%) had  some evidence of infection with hepatitis-B virus (H.B.V.), but the cases were more likely to be anti-HBc(+) and less likely to be anti-HBs(+) than the controls. (2) Most of the mothers of the cases were HBsAg(+) (71%), whereas only 14% of the mothers of controls were HBsAg(+). Lover titres of anti-HBs were less common in the mothers of the cases. (3) None of 27 fathers of cases had detectable anti-HBs, but 13 (48%) of the fathers of controls were anti-HBs(+). (4) Siblings of the P.H.C. cases were more likely to have anti-HBs than either their sibs with P.H.C. or the sibs of the controls. However, sibs of P.H.C. cases had lower titres of anti-HBs than the sibs of the controls. These data are consistent with the hypothesis that the P.H.C. cases were infected with H.B.V. by their mothers and that there was an environmental factor which affected the immunological response of all family members to H.B.V. Infection with H.B.V. and the mode of response to that infection among members of families appear to be major factors in the aetiology of P.H.C. in West Africa.


Oral zinc sulphate in rheumatoid arthritis. A preliminary trial of oral zinc supplementation was conducted in twenty-four patients with chronic, refractory rheumatoid arthritis. Zinc sulphate (220 mg three times daily) or placebo capsules of identical appearance were added to pre-existing therapy for 12 wk. This double-blind trial was followed by an open 12-wk period when all subjects took zinc. During the double-blind phase, zinc-treated patients fared better than controls with regard to joint swelling, morning stiffness, walking time, and the patient's own impression of overall disease activity. The indices and joint tenderness also improved with zinc treatment in both groups of subjects during the second 12-wk period. These encouraging results indicate that oral zinc sulphate deserves futher study in patients with active rheumatoid arthritis.


Correction of abnormal coagulation in chronic liver disease by combined use of fresh-frozen plasma and prothrombin complex concentrates. The effect on abnormal coagulation tests of infusions of fresh-frozen plasma (F.F.P), prothrombin complex concentrates, and a combination of these treatments was compared in 30 patients with  chronic liver disease undergoing needle biopsy. A single dose of F.F.P. (12 ml/kg body-weight) was found to be the least effective therapeutic regimen. The concentrate containing factors II, IX, and X was also not adequate, but the additional administration of factor-VII concentrate corrected the prothrombin-time (P.T.) and "Normotest" (N.T.) in most patients. However, this regimen did not correct the prolonged kaolin activated partial thromboplastin-time (K.P.T.T.). The results of tests for exploring both the extrinsic (P.T. and N.T.) and intrinsic (K.P.T.T.) coagulation systems only became normal after the combined administration of a lower dose of F.F.P. (8 ml/kg body-weight) and of both concentrates (12 units/ml). There was no clinical or laboratory evidence of thrombotic complications. No patient developed acute hepatitis or hepatitis-B surface antigen in the twelve months after biopsy. These results indicate that prothrombin-complex concentrates in combination with F.F.P. may therefore be used to allow liver biopsy to be performed safely in patients presenting with severe coagulation defects.


An augmented Schilling test in the diagnosis of pernicious anaemia. A patient with otherwise typical addisonian pernicious anaemia and serum anti-intrinsic-factor antibody failed to respond to oral intrinsic factor on repeated testing during three years of therapy with parenteral vitamin B 12. There was no evidence of generalised malaborption. To test the hypothesis that binding of intrinsic factor to gut secretory antibody was responsible, an "augmented" Schilling test was devised using eight times the usual dose of intrinsic factor. This increased dose of intrinsic factor resulted in normal absorption of the test dose of vitamin B 12 confirming the diagnosis. It is suggested that the augmented Schilling test may be useful in the diagnosis of the occasional patient with features of pernicious anaemia who fails to respond to conventional doses of intrinsic factor in the Schilling test.


Interaction of HLA molecules with non-immunological ligands as an explanation of HLA and disease associations. It is postulated that alloantigens (HLA in particular) may interfere with ligand/receptor interactions not directly involved in immune reactions. This interference may be regarded as a side-effect of the creation of new mutant HLA alleles which are maintained in the population for other reasons. As an example, it is suggested that some HLA molecules may have structures resembling those on receptors for certain hormones which could cause competition between HLA and receptor molecules for the hormone. Facilitation of ligand/receptor interaction may also be envisaged. Such interactions could under certain conditions lead to disease and might explain some associations between HLA and non-immunological diseases (e.g., manic-depressive disease and haemachromatosis).


History-taking for medical students. I-Deficiencies in performance. A videotape analysis of histories conducted by 50 senior medical students was carried out to assess their history-taking skills. As predicted, serious deficiencies (similar to those evident in inexperienced medical students) were found. It is argued that traditional methods of clinical training fail to equip medical students with adequate history-taking skills. They should be given a more appropriate history-taking scheme, the opportunity to practice this under conditions of direct observation within strict time-limits, and detailed feedback about their performance.


History-taking for medical students. II-Evaluation of a training programme. Two experiments designed to evaluate a programme for training medical students in history-taking skills were carried out. Results of the first indicate that students who underwent the programme reported almost three times as much relevant and accurate information after a test interview as those who received only traditional training. Results of the second experiment suggest that most of the programme's effect is attributable to discussion of a printed handout which presents the student with a detailed scheme for taking histories.


Dopamine and schizophrenia.

Dopamine and schizophrenia. The antipsychotic actions and extra-pyramidal side-effects of neuroleptic drugs are strongly correlated with their ability to block central dopaminergic transmission. It is argued that the former are more closely related to actions on dopaminergic mechanisms in the "mesolimbic dopamine" system, and the latter to similar actions in the striatum. Although the amphetamine psychosis closely resembles paranoid schizophrenia and may be due to excess dopamine release, clinical, biochemical, and endocrine studies suggest that dopaminergic overactivity is not a necessary concomitant of schizophrenic illnesses. It is suggested that the primary defect in schizophrenia does not lie in the dopamine neuron. It remains to be excluded that the receptors, particularly in the mesolimbic dopamine areas, become supersensitive, or that there is a deficit in a system which normally acts in antiagonism to the to the mesolimbic dopamine system.


[Experimental studies on the reaction of the immunological system during chronic otitis media and about the course of this disease (author's transl)]. During chronic otitis media intact immunoglobulins are split due to the proteolytic activity of extracellular bacterial proteinases into fragments of different molecular weight. The most malignant bacterial proteinases are the proteinases of pseudomonas aeruginosa (pyrocyanea). These proteinases can only be inhibited by alpha-2-macroblobulin of human blood serum. Because of its high molecular weight we find this inhibitor only in a very low concentration in the middle ear secretion. The destruction of the immunoglobulins is certainly one of the factors of weakening the immunological system in the middle ear. The inhibitory system with alpha-1-antitrypsin, inter-alpha-trypsin inhibitor and alpha-1-antichymotrypsin is unable to inhibit these bacterial proteinases of pseudomonas aeruginosa. The only possibility to get a high concentration of alpha-2-macroglobulin in the middle ear secretion is the liberation of this inhibitory by injuring blood vessels during a tympanoplasty. By this procedure the proteinases of pseudomonas aeruginosa with maximum activity at pH 7.8 and with a high proteolytic activity are almost completely inhibited. By blocking these proteinases combined with an appropriate antibiotic therapy and with the reconstruction of the destroyed parts of the middle ear by a tympanoplasty we can produce a preponderance of the immunological system as compared with the proteolytic activity of the proteinases. This high proteolytic activity can be a cause of the destruction of the small processes of the ossicular bones, especially of the lenticular process of incus. In order to demonstrate that there are proteolytic splitting processes of intact immunoglobulins, a quantitative analysis of the immunoglbulins IgG, IgA and IgM was done pre- and postoperatively. By these studies we found postoperatively a much higher level of intact immunoglobulins, particulary in cases of chronic otitis media associated with cholesteatoma. The blocking of the proteinases and the increase of the level of intact immunoglobulins combined with the reconstruction of physiological conditions in the chronically inflamed middle ear by a tympanoplasty lead to a stabilsation of the immunological and inhibitory system and create the prerequisites for a healing process in chronic otitis media. It is the purpose of further studies to learn about the capability of the split-products of the immunoglobulines to attach and to absorb antigens and toxins during a chronic inflammation in the middle ear.


[About some changes of the vocal cord mucosa and of Reinke's Subepithelial space (author's transl)]. The mechanical irritation alone or in combination with an inflammatory component and fragility of blood vessels plays the decisive role in the development of some vocal cord morphological changes. The duration of the mechanical irritation modifies the clinical picture. The plase of the strongest physiological mechanical irritation is the place of localization of the vocal cord morphological changes. The histological picture of different pathomorphological vocal cord changes is almost identical, this being the consequence of the specific structure of Reinke's subepithelial space. The treatment of all these morphological entities is surgical by means of microlaryngoscopy, except in cases of recent nodules which respond to conservative phoniatric therapy in the majority of cases.


[Subepithelial changes in simple leukoplakic hyperplasia of the laryngeal mucosa (author's transl)]. Subepithelial changes in hyperplasia of the laryngeal mucosa consist in edema and cellular inflammatory reaction. 150 cases of epithelial hyperplasia without atypia have been investigated histologically in regard to these subepithelial changes. The cases were subdivided into 3 categories, according to the degree of hyperplasia. The degree of inflammatory reaction in the subepithelial compartment also was classfied into 3 groups. A, B and C. There was evidence of a positive correlation between the increase in the epithelial hyperplasia and the increase of a positive correlation between the increase in the inflammatory process in the subepithelial layer. It could not be decided, which one of these two pathological changes was primary and which secondary. The therapy, however, should aim at both, epithelial proliferation, as well as, subepithelial inflammation.


Early assessment and prevention of sociocultural deprivation in infancy. The Bayley Infant Scales are used to diagnose early socio-cultural deprivation. It seems to be possible to observe early syndromes of pure environmental stress, as against syndromes of neurological damage, as well as syndromes of mixed traumatisation. There are also types of inadequate mothers in the low and lowest socio-cultural milieu who produce typical behavioral deficiencies in organically healthy infants. On the basis of a Scalogram Analysis of the Bayley Scales a structured stimulation technique has been designed which can be easily taught to and administered by professionals, semiprofessionals, volunteers and the parents themselves. Some case histories are presented to demonstrate the efficiency of the diagnostic and intervention procedures.


Development, validation and dissemination of instructional materials: an analysis. This paper describes the activities of SIMO (Self-Instructional Materials in Ophthalmology), a 2-year project to develop four slide/tape units and one videocassette in basic ophthalmology for medical students. The background of the project, some observations regarding pitfalls in the developmental process, data on product evaluation, and factors in effective dissemination are presented. Instructional effectiveness was documented using measures of knowledge, skill in recognizing abnormalities, and performance of diagnostic and management techniques.


The value of self-test items in tape--slide instruction. Two tape-slide sequences in general pathology were used in an experiment to assess the value of self-test items and to determine whether it is better to intersperse the self-test items in the programme or place them all at the end of the sequence. The programmes were presented to a random sample of thirty-six students from a class of 149 in one of three forms: version 1, tape-slide programme without self-test items; version 2, tape-slide programme with self-test items interspersed between sections of the sequence; and version 3, tape-slide programme with all self-test items at the end of the sequence. Each student worked a pre-test before studying versions 1, 2 or 3 of a programme. A week later they worked through the post-test which was identical to the pre-test. At the same time they filled in a short attitude questionnaire on the teaching method. All students learned from the programmes. There was improvement in the post-test on the mean pre-test scores for all versions of both programmes. For one programme there was no significant difference between the mean post-test scores for students studying versions 1, 2 or 3, but for the other programme there was a significant difference between the versions. In this case the inclusion of self-test items was better for learning than no self-test items, and it was better if the self-test items were placed at the end of the sequence. A highly favourable attitude to the method is reported.


Indium-111 bleomycin tumor scanning in lymphoma. A new radiopharmaceutical, indium-111 labeled bleomycin (IB), was evaluated as a tumor-imaging agent in 55 patients with lymphoma. Overall disease activity was correctly identified in 79% of 75 whole-body scan obtained 48 hours after intravenous administration of IB. Serial scans in 19 patients accurately reflected changes in their disease status. Lymphatic and soft tissues  sites of involvement both above and below the diaphragm were most readily identified by scanning. Bone marrow and hepatic involvements were more difficult to detect because of normal tissue background in the considered organs. Five patients manifested diffuse pulmonary uptake of IB and only in one case was it explained. To determine the accuracy of scanning with IB, we evaluated 731 individual sites of potential tumor involvement with these results: true positives 90%, false negatives 10%, false positives 4%, and true negatives 96%. Thus, tumor scanning with indium-111 bleomycin is an important new technique for the initial staging and serial evaluation of patients with lymphoma.


Serum vitamin B12-binding proteins in a case of eosinophilic leukemia. A patient with subacute eosinophilic leukemia is presented, with full recognition of the controversy surrounding that entity. Serum vitamin B12 and B12-binding protein studies and simultaneous complete blood counts were done before and during 6 months of high-dose, intermittent combination chemotherapy. The patient presented with extremely high levels of serum vitamin B12, unsaturated B12-binding capacity, and transcobalamin I, all of which resembled the highest values seen in chronic myelogenous leukemia. Serial studies, during and after remission induction, showed a precipitous fall of serum vitamin B12 and unsaturated B12-binding capacity to normal levels. The data show that transcobalamin I levels, which eventually reached low-normal range, correlate best with the level of circulating and bone marrow eosinophils. Transcobalamin II and serum third binder appeared to be normal throughout the patient's course. The B12-binding protein abnormalities are not considered diagnostic of eosinophilic leukemia.


[Inoculation for tourism (author's transl)]. The inoculations prescribed or recommended by the authorities responsible on the basis of the International Health Regulations for international tourism are first discussed and the contraindications gone into. Then some inoculations or measures for specific prophylaxis are presented for which there are no international regulations, but which have a particular significance from the point of view of tourism. These are inoculations against poliomyelitis, tetanus, typhoid fever and prophylaxis of hepatitis A with conventional gamma globulin.


[Hepatitis: an international problem (author's transl)]. Three kinds of virus hepatitis are recognized today: hepatitis A, B, and "non A-nonB". Hepatitis A is transmitted mainly by the anal-oral route, hepatitis B and probably also the third form of hepatitis principally by direct inoculation or close physical contact. Normal human immune serum globulin protects against hepatitis A, but only gives limited protection against hepatitis B and "non A-non B" hepatitis. Special immune serum globulin provides better protection but it is only available in small quantities and should be reserved for direct inoculation only. Vaccines for active immunization against hepatitis A and "non A- non B" hepatitis have not yet been developed and active immunization against hepatitis B with HBs-Ag is still in the experimental stage.


[Incidence of viral hepatitis in travelers in the tropics. Experiences with gamma globulin prophylaxis (author's transl)]. The occurrence of virus hepatitis during their residence was determined in 2755 persons returning from the tropics in 1975. The incidence was particularly high in those returning from India, Pakistan and Afghanistan. There was also a greater risk of the disease for travelers in Ethiopia, the countries on the West coast of Africa and some South American States. On the average, persons returning form the tropics fell ill 10 times more frequently with virus hepatitis during their residence in the tropics than in the Federal Republic. Particularly endangered occupational groups were the staffs of the health services and the social workers. Immunization with gamma globulins prevented an attack of virus hepatitis in all cases for a period of 4-6 months. Later, the effect of the immunization could no longer be established.


[Degradation of 14C-lindane by a mold culture]. The metabolism of 14C-lindane, its short-lived main metabolite gamma-2,3,4,5,6-pentachlorocyclogexene and penta-/hexachlorobenzene was followed in a mould culture after 6, 17, 30 and 52 days. The distribution in the culture medium was also investigated.


[In vitro behavior of 14C-lindane in blood]. Almost 55% of lindane (gamma-HCH) which had been applied in vitro are removed by simple hexane extraction of the blood; if blood fractions are extracted, a total rate of 95% results. The partition of gamma-HCH between the plasma and the erythrocytes is approximately I:I; when HCH erythrocytes are incubated in fresh plasma, almost 60% of erythrocyte HCH pass into the plasma. The erythrocytes absorb nearly 90% of the HCH contained in an aqueous medium. Haemolysis showed that HCH is most probably transferred through the membrane. The partition of HCH between the cytoplasm and the membrane fraction ranges from 3:7 to 7:3. TCA precipitates almost 90% of erythrocyte HCH together with the protein. The absorbing capacity of the erythrocytes could be followed up to nearly 0.4-10(-6)M/ml of blood. Cadmium increases, lead decreases the binding of HCH to the erythrocytes; mercury and ethanol exert but small effects.


[Lead content of vegetable foods in vicinity of lead emitting industry]. By means of atomic absorption spectrophotometry, the authors determined the lead content in vegetable, fruit and soil samples from the environments of a lead-smelting plant. The lead values obtained were some 10- to 100-fold higher than those found normally. The mean lead content in vegetables varied from 2 to 150 p.p.m. in dry matter; that in fruits, from 4 to 35 p.p.m. Lead concentrations ranging from 575 to 2900 p.p.m. were detected in the soil samples. The mean lead content in the atmosphere was between 0.62 and 0.95 mug/m3 (maximum value 12 mug/m3). From these data it was calculated that the total resorption of lead from vegetables, fruits, potatoes and air varied from 0.04 to 0.49 mg/day in the territory under investigation. To protect the population against increased intake of lead, it is recommended to abandon individual and commercial vegetable and fruit growing, especially leaf vegetables. Since the contamination of the soil persists for generations, the authors discuss its replacement by non-contaminated surface soil.


gag Gene of mammalian type-C RNA tumour viruses. The translation product of the gag gene of mammalian type-c- RNA viruses is a 65,000-68,000 molecular weight precursor polypeptide (Pr65) whose cleavage leads to the formation of four virion proteins, p30, p15, p12 and p10. An immunological approach has been used to establish the arrangement of the sequences coding for these proteins within the viral genome as (5') p15-p12-p30-p10 (3').


[Formation of cellular structures and interneuronal connections in an organotypic culture of newborn mouse hippocampal tissue]. The cytoarchitectonic structures and fibres pathways of the newborn mouse hippocampus were studied in vitro. After 2-3 weeks in vitro it was possible to see on transverse slices of the hippocampus main fascicles (alvear tract, perforant path, axons of granular cells) and cytoarchitectonic formations (pyramidal cell layer of hippocampus, infra- and supra-pyramidal lymbs of dentate gyrus granular cells). The postnatal processes of histogenesis are suggested to result in subsequent differentiation of the cytoarchitectonic formations and in the systemic growth of nerve fibres in the hippocampus explants. Thus, organo-typic explants of the hippocampus represent a useful model systems for electrophysiological studies in systemic interneuronal connections formed de novo as a result of postnatal development in vitro.


The development of the blood-CSF barrier after birth. The serum/CSF concentration ratio of alpha-2-macroglobulin is used as parameter to follow the development of the blood-CSF barrier throughout childhood. The decline of the protein permeability within the first year of life has all characteristics of a maturation process. The tightening of the barrier is thought to be the last stage of a continuous developmental process of capillaries. It is accompanied by a compositional shift in favour of small molecular serum proteins.