Action of endotoxin on lymphoid cells.

Action of endotoxin on lymphoid cells. Endotoxin (lipopolysaccharide, LPS) has been found to act on all three cell types of the immune system, thymus-derived (T-) cells, bone marrow-derived (B-) cells, and macrophages. LPS is mitogenic for B-lymphocytes and activates them to release a chemotactic lymphokine. Macrophage activation appears to be mediated by macrophage-activating factor, another lymphokine released from B-cells. In addition, LPS acts synergistically with phytohemagglutinin to initiate division of purified T-lymphocytes. All these phenomena are mediated by the lipid A moiety of LPS. The role of lymphoid cells in mediating the lethal effects of LPS have also been investigated. The adoptive transfer of spleen cells from LPS-responsive mice (C3H/HeN) to LPS-resistant but histocompatible mice (C3H/HeJ) rendered the LPS-resistant mice significantly more susceptible to LPS-induced lethality. These findings suggest that spleen cells play an essential role in mediating the lethal effects of LPS in vivo.


Antibody of polyribophate of Haemophilus influenzae type b in infants and children: effect of immunization with polyribophosphate. Antibody to polyribophosphate, the capsular polysaccharide of Haemophilus influenzae type b, was measured in healthy ambulatory children by a radioactive antigen-binding assay. Titers fell from birth through nine months of age, then increased until six years, when they plateaued. Antibody activity was not correlated with the child's sex, ethnic status, or area of residence. Doses of 0.2-50 microgram of polyribophosphate given as single or booster doses had similar effects on antibody activity. Of 368 doses given to infants two to six months of age, 7% produced a significant antibody response; of 95 doses given to infants seven to 12 months old, 17% produced a response. The geometric mean titers of antibody resulting from immunization with polyribophosphate given at various times in relation to diphtheria-pertussistetanus vaccine did not differ significantly from one another or from titers observed in infants given only the latter vaccine. These data indicate that purified polyribophosphate will not provoke humoral immunity in young infants against H. influenzae type b and that it should no longer be considered as a candidate vaccine for this purpose.


Immunogenicity in weanling rabbits of a polyribophosphate complex from Haemophilus influenzae type b. Polyribophosphate (PRP), the capsular polysaccharide of Haemophilus influenzae type b, is more effectively immunogenic when it is associated with the bacterium than when it is in the purified form that is being tested as a vaccine for humans. In an effort to analyze this difference, we isolated from H. influenzae type b a high-molecular-weight, soluble complex, in which PRP appears to be combined with protein (about 7% protein). The pyrogenicity and limulus lysate gelation activity of the complex suggest that a small amount of lipopolysaccharide also is present. The protein was resolved into five polypeptides by electrophoresis in polyacrylamide gel containing sodium dodecyl sulfate. In weanling rabbits, which do not respond to purified PRP, the complex induces high titers of antibody of PRP, in an anamnestic pattern. Bactericidal antibody to other bacterial components was also elicited. Equilibrium density gradient centrifugation of the complex indicated that most of the immunogenicity of PRP resides in the least dense fractions, which are high in protein, low in polysaccharide, and active in the limulus lysate test; denser fractions that react strongly with limulus lysate but are poor in protein were much less immunogenic.


Lymphocyte response to phytohaemagglutinin before and after radiation therapy in patients with carcinomas of the head and neck. The ability of peripheral blood lymphocytes to respond in vitro to phytohaemagglutinin (PHA) was studied before and after radiation therapy in patients with locally advanced squamous cell carcinomas of head and neck. Patient lymphocyte response to PHA was depressed before therapy and further declined following therapy. The pre-therapy PHA responsiveness was found to be prognostically significant in regard to patient survival.


Management of seven stable levels of brain death. We have presented the choices available in the management of seven representative kinds of brain death and partial death as well as their legal ramifications. Refinements of tests of brain functions and new methods of evaluating cerebral blood flow allow more and more accurate estimates of brain viability. Human qualities of life are directly related to how much and which parts of the brain are viable and are indistinguishable from life itself. The decision to end life support should at present remain with the attending physician. New statutes concerning brain death imply that irrefutable technical evidence is readily available to diagnose brain death, that brain death is as valid a sign of death as any former criteria, and that in certain situtations brain death must be used to pronounce death. From the medical practitioner's viewpoint, we believe that a brain death law would rarely be useful and that community and national customs concerning heroic efforts would be sufficient. We do support a simple acknowledgement by statute that brain death may be a cause for determining death. The legal need for determining a precise time of death is understandable, but premature and restrictive definitions based on a rapidly evolving technology will create many problems.


Effects of angiotensin II on arginine-vasopressin in physiological and pathological situations in man. Studies were designed to determine whether angiotensin II has a direct stimulatory effect on arginine-vasopressin in man and to determine the role, if any, played by angiotensin II in the control of vasopressin release in physiological and pathological conditions. Acute infusion of angiotensin II in normal volunteers produced small but definite increases in plasma levels of arginine-vasopressin (5-4+/-0-3(S.E.M.) to 6-4+/-0-2 pg/ml) only when plasma angiotensin II levels were supraphysiological. Concurrent measurements of plasma arginine-vasopressin and angiotensin II were made during acute changes in fluid balance and posture in normal volunteers and in clinical conditions characterized by high plasma levels of angiotensin II (Addison's disease and Bartter's syndrome). The results of these studies allow us to conclude that there is little to suggest a direct effect of angiotensin II which is likely to be relevant to the normal physiological control of arginine-vasopressin in man.


Induction and properties of cytoplasmic factor(s) which enhance nuclear nonhistone protein phosphorylation in lymphocytes stimulated by anti-Ig. An increased in vitro phosphorylation of nonhistone nuclear proteins (NHP) was observed in the nuclei isolated from rabbit lymphocytes which had been stimulated with anti-Ig for 4 h. No concomitant increase of phosphorylation in histones or 0.14 M NaCl-soluble proteins was observed. The increase of in vitro phosphorylation of NHP was also observed in the nuclei isolated from nonstimulated cells when these nuclei were preincubated for 2 h with cell-free extracts from anti-Ig-stimulated cells. The active substance in cell-free extracts was maximally induced when lymphocytes were stimulated with anti-Ig for 2 h. The induction of an increased phosphorylation of NHP in nonstimulated nuclei with the cell-free extracts was not due to decrease of the adenosine triphosphate pool in the extracts from anti-Ig-stimulated cells. The active substance in cell-free extracts was not NHP-protein kinase itself, but it probably activated NHP-protein kinase in quiescent nuclei. The active substance was nondialyzable and probably protein. It was resistant against heating at 56 degrees C for 30 min, but the activity was completely destroyed by heating at 90 degrees C for 30 min. The active substance may be responsible for the transduction of the membrane-mediated signals given through Ig receptors to nuclei.


Immunologic recognition of influenza virus-infected cells. II. Expression of influenza A matrix protein on the infected cell surface and its role in recognition by cross-reactive cytotoxic T cells. Two distinct subpopulations of cytotoxic T cells are generated in the primary or secondary response of mice to type A influenza viruses. One subpopulation is specific for the immunizing virus strain. The other subpopulation shows a high degree of cross-reactivity for heterologous type A virus of a different subtype. This report examines the possibility that distinct influenza virus antigens, expressed on the surface of the infected cell, are recognized by the different subpopulations of influenza-specific cytotoxic T cells. Data are presented which demonstrate that influenza A matrix protein, an internal virion antigen, is detectable on the surface of target cells infected with influenza A viruses of different subtypes. Since this viral antigen is type specific, i.e., serologically cross-reactive among all type A influenza viruses, it could serve as the target for cross-reactive cytotoxic T cells. To further examine the specificity of the two cytotoxic T-cell subpopulations, experiments were carried out by using the inhibitor of glycoprotein synthesis - 2-Deoxy-D-Glucose 2-DG. These experiments examine first the effect of 2-DG on the expression of influenza matrix protein and viral glycoprotein on the infected cell surface and second, the susceptibility of 2-DG-treated target cells to lysis by cytotoxic T cells. 2-DG inhibits the expression of the viral hemagglutinin glycoprotein on the cell surface but does not inhibit the expression of the nonglycosylated matrix protein. Furthermore, inhibition of glycoprotein synthesis in infected target cells abrogates the reactivity of infected target cells to lysis by virus strain-specific but not cross- reactive cytotoxic T cells. These findings suggest that the influenza glycoproteins (hemagglutinin and/or neuraminidase) and the nonglycosylated matrix protein are the targets for the virus strain- specific and cross-reactive cytotoxic T cells, respectively. These results are discussed in the light of available information on influenza virus structure and the biology of influenza infection and in terms of current models for cytotoxic T-cell recognition of virus-infected cells.


Energy metabolism and T-cell-mediated cytolysis. I. Synergism between inhibitors of respiration and glycolysis. The energy requirements for T-cell-mediated cytolysis have been investigated. Cytolytic thymus-derived lymphocytes (CTL) were generated in vitro in mixed leukocyte cultures and assayed for cytotoxicity on 51Cr-labeled mastocytoma target cells. Cytolysis was only slightly reduced in the absence of exogenous glucose (less than 5 micrometer) or under conditions of extreme hypoxia (less than 0.2 micrometer oxygen). Furthermore, neither the glucose analogues 2-deoxy-D-glucose and 5-thio-D-glucose nor the respiratory antagonists sodium azide and 2,4-dinitrophenol were very effective inhibitors of cytolysis when used individually. However, these glucose analogues were highly effective in inhibiting cytolysis in the absence of oxygen, and the respiratory antagonists inhibited cytolysis to a much greater extent in the absence of glucose. In addition, synergistic effects were observed when the glycolytic and respiratory inhibitors were combined. Taken together, these results indicate that T-cell-mediated cytolysis is an energy-dependent process which can be supported by either oxidative or glycolytic energy pathways.


Energy metabolism and T-cell-mediated cytolysis. II. Selective inhibition of cytolysis by 2-deoxy-D-glucose. The effect of the hexose analogue 2-deoxy-D-glucose (2-DG) on T-cell-mediated cytolysis has been investigated. 2-DG inhibited cytolysis in glucose-free medium but not in medium containing equimolar concentrations of glucose. This inhibition was reversible and quantitatively competitive with glucose. Among other natural sugars examined, only mannose competed effectively with 2-DG and reversed the inhibition of cytolysis, whereas sodium pyruvate, fructose, galactose, fucose, and glucosamine were without effect. Mannose and glucose were equally effective in competing with 2-DG on a molar basis. When other glucose analogues such as 5-thio-D-glucose (5-SH-G) and 3-O-methylglucose were investigated under the same conditions, no inhibition of cytolysis was observed however, 5-SH-G (but not 3-O-methylglucose) was able to reverse the inhibitory effect of 2-DG in a competitive fashion. Taken together with the data presented in the accompanying paper, these findings provide strong evidence that 2-DG inhibits T-cell-mediated cytolysis by a mechanism that is unrelated to energy production. The possibility that inhibition is related to interference with membrane glycoprotein synthesis is discussed.


Separation of helper and suppressor T lymphocytes. II. Ly phenotypes and lack of DNA synthesis requirement for the generation of concanavalin A helper and suppressor cells. Using a Ficoll velocity sedimentation gradient, we have been able to fractionate concanavalin A (Con A)-induced helper and suppressor cells into separate pools. Cells activated by Con A to mediate helper activity are Ly1+, do not require DNA synthesis for induction, and remain as small cells after activation. Suppressor cells are Ly23+, are found in the blast cell fraction and their induction is not inhibitable by prior treatment with mitomycin C or irradiation, both of which inhibit DNA synthesis. The implications of such findings are discussed.


Inhibition of T-antigen-binding cells by idiotypic antisera. Shared idiotypy between B- and T-cell receptors specific for the antigen L-tyrosine-p-azophenyltrimethylammonium [tyr(TMA)] was studied in an antigen-binding assay using idiotypic antisera. These idiotypic reagents were prepared by inoculation of rabbits with purified anti-tyr(TMA) antibody raised in strain 13 guinea pigs. The antisera blocked 78-83% of the antigen-binding T cells (T-ABC) and 50-55% of the antigen-binding B cells (B-ABC) from tyr(TMA)-immune strain 13 and outbred lymph node cells (LNC). An excess of normal guinea pig Ig in the ABC assay did not affect the ability of the idiotypic antisera to block T- and B-ABC. Nylon wool-passed tyr(TMA)-immune LNC were trypsin treated resulting in a 75% loss of T-ABC. The trypsin-treated population was then cultured for 16 h which resulted in a return of T-ABC to 92% of pretrypsin values. 77% of these regenerated T-ABC could be blocked with idiotypic antisera. Specificity of the idiotypic antisera was tested in L-tyrosine-p-azobenzenearsonate-immune guinea pig LNC. Neither T- nor B-ABC were blocked in this heterologous system. Further blocking experiments were performed to characterize the nature of the T-ABC receptor. A variety of anti-Ig reagents, some of which block B-ABC, do not inhibit T-ABC suggesting that variable regions on T cells are not linked to Ig Constant regions.


Cell-mediated mitogenic response induced by leukoagglutinin and Lens culinaris lectin in mouse lymphocytes. The proliferative response of mouse lymphocytes to syngeneic cellular stimulation upon membrane modification with lectins was studied. Brief pretreatment of stimulator cells (mitomycin-C-treated spleen cells) followed by mixed culture with syngeneic cortisone-resistant thymocytes resulted in a significant proliferative response in the thymocytes. This effect was not due to a soluble mediator and was similar to the mitogenic response after Con A-induced membrane modification reported previously. Because of its general characteristics, we refer to this response as cell-mediated mitogenic response (CMMR). Cell contact between stimulator and responder cells was necessary but not sufficient for the induction of the response. The lectins that generated CMMR were T-cell mitogens. CMMR was generated in all the syngeneic combinations tested and even in allogeneic combinations. No detectable cytotoxic activity towards syngeneic targets cells was produced after CMMR. Moreover, CMMR in allogeneic combinations led to the suppression of the generation of specific cytotoxic lymphocytes. Population analysis with antibodies against T or B cells, nylon wool fractionation of stimulator cells, and tests with peritoneal macrophages and with spleen cells from athymic mice revealed that CMMR depends predominantly on the interaction between responder T cells and stimulator Ig-positive lymphocytes.


Ontogeny of cell-mediated immunity. I. Early development of alloantigen-specific cytotoxic T-cell precursors in postnatal mice. Cytotoxic T-cell precursors have been shown to occur in spleens of 2-3-day-old mice. By 12 days after birth, the cytotoxic T-cell response of spleen cells to alloantigens has reached 23-32% of adult levels. Addition of extra T-helper cells did not permit cytotoxic T-cell development in spleen cells from newborn to 2-day-old mice suggesting either a lack of precursors or suppression of precursors. The ontogeny of cell-mediated immune functions has thus been shown to correlate well with other work on the development of humoral immunity, accessory cells, and graft versus host reactivity.


Hemagglutinin-specific cytotoxic T-cell response during influenza infection. Specific cytotoxic thymus-derived (T) lymphocytes were detected in the cervical lymph nodes and spleen during influenza infection of mice. The cytotoxic T cells can distinguish target cells infected with different influenza A subtypes. Infection with parent viruses and their recombinant progeny possessing the  hemagglutinin of one parent and the neuraminidase of the other demonstrated that significant cytotoxicity occurred only when the hemagglutinin of the immunizing viruses was the same as that of the virus used to infect the target cell. In addition to this specific cytotoxic response to the major surface antigen, a cross-reactive response could be detected when the relatively nonpermissive L cell was used as the target cell. These results indicate there is a specific cytotoxic T-cell response to the surface hemagglutinin, and a cross-reactive cytotoxic response, not directed to the hemagglutinin, during influenza infection. The cytotoxic T-cell response specific for the hemagglutinin antigen may play an important role in in vivo immunity to influenza.


In vitro analysis of allogeneic lymphocyte interaction. I. Characterization and cellular origin of an Ia-positive helper factor-allogeneic effect factor. A soluble allogeneic effect factor (AEF) was produced by using H-2 congenic mouse strains and a serum.free cell culture medium. An AEF derived from untreated activated responder cells and irradiated stimulator cells provided helper cell function in a primary and secondary antibody response for both T-cell-depleted responder B cells and stimulator B cells. This interaction may be determined by genes situated in the I-A and I-B regions: additional K-region control was not excluded. Ia antigens, but neither H-2 nor Ig determinants are molecular constituents of AEF. The active components of this AEF consist, in part, of Ia antigens derived from both the activated responder cell population and irradiated stimulator cell population. An AEF derived from Ia negative responder cells and irradiated T-cell- depleted stimulator cells helps a secondary antibody response of T-cell- depleted stimulator B cells but not responder B cells. This genetically restricted AEF contains Ia antigens determined by the stimulator haplotype but not the responder haplotype. The priming antigen, DNP- keyhole limpet hemocyanin, is not a component of restricted AEF. The data suggest that restricted AEF may be a product of a stimulator B cell and/or macrophage. They support the hypothesis that the recognition by allogeneic T cells of Ia antigens on B cells activates the B cell to IgG antibody production.


Induction of specific immune unresponsiveness using purified mixed leukocyte culture-activated T lymphoblasts as autoimmunogen. I. Demonstration of general validity as to species and histocompatibility barriers. Normal immunocompetent T lymphocytes can be induced into specific proliferation if confronted with the relevant alloantigen in vitro. Such mixed leuko-cyteculture-activated T lymphoblasts carring idiotypic receptors on their surface can be purified using velocity sedimentation and serve as immunogen if administered in adjuvant to the autologous host. Autoblast immunization can be shown to lead to specific, long-lasting unresponsiveness against the relevant alloantigens, while leaving reactivity against third-party antigens intact. When tested as to general validity, it could be shown to function in all species analyzed (mouse, rat, and guinea pig) as well as across both major and minor histocompatibility barriers. No negative side effects have been noted so far. It would thus seem clear that autoblast immunization using the above described scheme may serve as a general tool in inducing long-lasting, specific unresponsiveness in any species and across any histocompatibility barrier.


Inability of mice with a defect in B-lymphocyte maturation to respond to phosphorycholine on immunogenic carriers. Mice with the CBA/N defect are unresponsive to the hapten phosphorylcholine (PC) even when presented on a variety of immunogenic carriers. Since these mice have the variable region gene for PC, their inability to respond may reflect deletion or suppression of the line of B lymphocytes which is responsible for the anti-PC response.


Antigens expressed by human B lymphocytes and myeloid stem cells. Antisera prepared against papain-digested spleen cell membranes were known to by cytotoxic for normal and neoplastic human B lymphocytes and for a majority of acute and chronic myeloid leukemic cells. It is now shown that these antisera are also cytotoxic for normal myeloid stem cells (CFU-C), thus providing a probable explanation for their occurrence in myeloid neoplasia.


Expression of phosphorylcholine-specific B cells during murine development. The TEPC 15 (T15) clonotype, a putatively germline antibody specificity, does not appear in the neonatal B-cell repertoire until approximately 1 wk of age. This report extends this observation by the demonstration that (a) the T15 clonotype follows similar kinetics of appearance in germfree as well as conventionally-reared mice; (b) maternal influences and genetic background play a minor role in the development of the T15 clonotype since CBFI neonates raised by C57BL/6 or BALB/c mothers acquire the T15 clonotype at the same time in ontogeny as BALB/c neonates; (c) the lack of phosphorylcholine (PC)-specific B cells shortly after birth is reflected in a dearth of PC-binding cells in the neonate as well; and (d) no PC-specifc B cells are found in 19-day fetal liver or in bone marrow until 7 days of life, coincident with their appearance in the spleen. These findings, along with a previous report that PC-specific splenic B cells are tolerizable as late as day 10 after birth, confirm the invariant, late occurrence of the T15 clonotype and support a highly- ordered, rigorously predetermined mechanism for the acquisition of the B- cell repertoire. The results are discussed in light of other studies on the ontogeny of B-cell specificity, and in terms of the implications on the mechanism by which antibody diversity is generated.


Immune response in humans after vaccination with vaccinia virus: generation of a virus-specific cytotoxic activity by human peripheral lymphocytes. After vaccinia virus vaccination of human volunteers, local indurations developed within 10 days, and regional adenopathy was detected in half of the individuals. Their peripheral blood lymphocytes (PBL) harvested at different days after vaccination showed specific activity against target cells infected with vaccinia virus with a peak activity at day 7. The specificity of the cytotoxic activity was not related to HLA markers, since autologous, homologous, and heterologous infected target cells were lysed with the same efficiency. The cytotoxic activity was caused by PBL that did not rosette with sheep erythrocytes and could be depleted by more than 90 percent by removing Fc receptor-bearing cells. T-cell- depleted PBL showed a one-half to two times greater cytotoxicity than intact PBL. The cytotoxic activity could also be abrogated by more than 95 percent by rabbit Fab(2) anti-human IgG. On the other hand, nonimmune PBL lysed vaccinia-infected target cells in the presence of specific antibodies against vaccinia virus, thus demonstrating that ADCC could be efficient in lysing vaccinia-infected target cells. We conclude that after vaccination, antibody-forming cells arise and provide specific anti-viral antibody and that the cytotoxic cells detected in this reaction are K cells. These experiments suggest that antibody-dependent cell cytotoxicity may be of major importance in the recovery of man to virus infections.


Immunosuppressive factor(s) specific for L-glutamic acid50-L-tyrosine50 (GT). III. Generation of suppressor T cells by a suppressive extract derived from GT-primed lymphoid cells. Injection of mice with L-glutamic acid50-L-tyrosine50 (GT)- or L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT)-specific suppressor T-cell factor (GT-TsF or GAT-TsF) up to 5 wk before antigenic challenge challenge suppresses GT-methylated bovine serum albumin (MBSA) and GAT-MBSA plaque-forming cells responses. T suppressor cells are responsible for the suppression induced by the suppressive extract as demonstrated by adoptive transfer and sensitivity to anti-Thy-1 and complement treatment. We conclude that suppressive extract induces specific suppressor T cells. The material responsible for generation of suppressor T cells is a product of the I subregion of the H-2 complex. We have excluded that suppressive quantities of antigens are present in the extract. A/J mice, which can neither be suppressed by GT nor make GT-TsF can be suppressed by BALB/c GT-tsf. Spleen cells from BALB/c GT TsF-primed A/J mice can adoptively transfer suppression to normal syngeneic recipients. A/J mice appear to be genetically defective in cells involved in factor production. These results are discussed in the light of a two-step model for induction of antigen-specific suppressor cells.


Isolation and characterizaiton of murine cell surface components. I. Purification of milligram quantities of Thy-1.1. The Thy-l.1 molecule was isolated from the BW5147 murine lymphoblastoid cell line. The initial step in purification was the preparation of a crude plasma membrane fraction followed by acetone precipitation. The acetone pellet was solubilized using deoxycholate (DOC) and Thy-1.1 was purified by use of a Lens culinaris lectin affinity column and an AcA-34 gel filtration column. The purified glycoprotein with Thy-1.1 activity had a mol wt of approximately 25,000 daltons. The isolation of this molecule was effected by detecting Thy-I activity utilizing rabbit anti- mouse brain serum tested on rat thymocytes. Congenic anti-Thy-l.1 serum was ineffective in detecting Thy-l.1 after DOC solubilization. An antiserum prepared in rabbits to the purified Thy-1.1 was found to be cytotoxic to mouse and rat thymocytes. The cytotoxic activity of this antisera could be completely absorbed with AKR/Jax brain and thymus but was not absorbed by liver. In addition, AKR/Jax thymocytes totally absorbed all cytotoxic activity of the rabbit anti-purified Thy-1 serum for BW5147 cells suggesting that the cell line shares identical specificities with normal thymocytes. The purified Thy-1.1 molecule was able to totally absorb the cytotoxic activity of mouse congenic anti-Thy-1. These studies serve as a model for the isolation of other murine lymphoid cell surface components in quantities for detailed structural and functional analysis.


Effects of strychnine on the potassium conductance of the frog node of Ranvier. The nature of the block of potassium conductance by strychnine in frog node of Ranvier was investigated. The block is voltage-dependent and reaches a steady level with a relaxation time of 1 to several ms. Block is increased by depolarization or a reduction in [K+]O as well as by increasing strychnine concentration. A quaternary derivative of strychnine produces a similar block only when applied intracellularly. In general and in detail, strychnine block resembles that produced by intracellular application of the substituted tetraethylammonium compounds extensively studied by C.M. Armstrong (1969. J. Gen Physiol. 54:553-575. 1971. J. Gen. Physiol. 58:413-437). The kinetics, voltage dependence, and dependence on [K+]O of strychnine block are of the same form. It is concluded that tertiary strychnine must cross the axon membrane and block from the axoplasmic side in the same fashion as these quaternary amines.


Effects of strychnine on the sodium conductance of the frog node of Ranvier. Strychnine blocks sodium conductance in the frog node of Ranvier. This block was studied by reducing and slowing sodium inactivation with scorpion venom. The block is voltage and time dependent. The more positive the axoplasm the greater the block and the faster the approach to equilibrium. Some evidence is presented suggesting that only open channels can be blocked. The block is reduced by raising external sodium or lithium but not impermeant cations. A quaternary derivative of strychnine was synthesized and found to have the same action only when applied intracellularly. We conclude that strychnine blocks sodium channels by a mechanism analogous to that by which it blocks potassium channels. The potassium channel block had previously been found to be identical to that by tetraethylammonium ion derivatives. In addition, strychnine resembles procaine and its derivatives in both its structure and the mechanism of sodium channel block.


Cellular mechanisms in the protection against infection by Listeria monocytogenes in mice. Listeria monocytogenes, in doses of 2-0 X 10(3) to 3-0 X 10(3) viable organisms, was injected into athymic nude mice, irradiated mice and mice treated with reticuloendothelial system-blocking agents. Viable counts on liver and spleen homogenates were made at intervals after infection. In both nude mice (nu/nu) and normal littermates (nu/+) of BALB/c background, the bacteria grew rapidly for 24 h but increased only slowly thereafter, to reach a plateau of about 10(5) per organ at 72 h. In nu/+ mice, the number of viable bacteria began to decrease after 6 to 9 days, with complete elimination by day 12. In nude mice, the number of Listeria remained at a stable level of approximately 10(5) per organ during the observation period of 21 days. In lethally irradiated nu/+ mice, bacteria grew progressively and extensively to reach 10(7) per spleen and 10(9) per liver by 72 h. Bacterial growth during the first 72 h was markedly enhanced by treatment with carbon particles, dextran sulphate 500 or silica. These enhancing effects were also observed in nude mice and in AKR, C3H/He and C57BL/6 animals. We conclude that both non-immune phagocytes and T cell-dependent mechanisms contribute to the resistance of mice to Listeria infection.


Treatment of fatal disseminated vaccinia virus infection in immunosuppressed mice. Studies were performed to compare the therapeutic effectiveness of three antiviral drugs (ARA-A, ARA-C and IDU) on the course of fatal disseminated vaccinia virus infection in immunosuppressed mice. Treatment with ARA-A begun as late as 7 days after virus infection was significantly effective in preventing death; no antiviral effect of the other two drugs was demonstrated.


Interstitial pneumonitis in the immunologically suppressed child: an urgent surgical condition. Thirty-eight children were evaluated for interstitial pneumonia by open lung biopsy. In most instances the patients were immunosuppressed as a result of cancer chemotherapy and irradiation. Pneumocystis carinii infection was the most common cause of pneumonitis (60.4%), especially in children with leukemia (78.3%). The clinical triad of hypoxemia, tachypnea, and a diffuse interstitial infiltrate on chest x-ray, is an indication for early open lung biopsy. Survival was 91.7% in cases of acute pneumocystis pneumonia, a significant improvement over previous reports. These observations strongly support the concept of early open lung biopsy in the management of diffuse interstitial pneumonitis in patients who are immunosuppressed.


Surgical complications of Ehlers-Danlos syndrome in children. The Ehlers-Danlos syndrome is a special challenge to the surgeon because of its relative infrequency and the late onset of overt symptoms, such as hyperelastic skin. We should consider this diagnosis in every child with recurrent inguinal herniae and search for its stigmata. A child with a known Ehlers-Danlos syndrome who requires an elective operation should be carefully studied for coagulation disorders. If there is a history of past injuries or operations in which there was poor healing, the benefits of the proposed operation must be carefully weighed against possible complications. Operations must be carried out with great care to avoid tearing the skin and fascia. Meticulous attention is given to hemostasis to prevent postoperative bleeding or hematomas. Inguinal hernias in these children resemble those seen in elderly patients. The transversalis fascia is thin and the internal ring is hugely dilated. An adult-type repair, with the use of mesh or felt may result in a lower incidence of recurrence. In addition, the Ehlers-Danlos syndrome may be yet another etiology for obscure gastrointestinal bleeding in children. Specific enzyme defects have been identified in 2 of the 7 subtypes of this disease. Further, biochemical study of connective tissue defects may contribute to our knowledge of normal collagen synthesis and wound healing.


Electron microscopic study of the orbital lesion of Hand-Schüller-Christian disease. The orbital bone lesion of a patient with Hand-Schüller-Christian disease was examined by electron microscopy. Vacuoles in the cytoplasm of foam cells contained granules which were shown by histochemical examination to be cholesterin. Langerhans cell granules in the histiocytes of Hand-Schüller-Christian disease showed a close resemblance to those in Langerhans cell of the epidermis, eosinophilic granuloma and Letterer-Siwe disease. Charcot-Leyden crystals appeared to be formed in the lysosomes of histiocytes in Hand-Schüller-Christian disease. From histopathological and cytological points of view, eosinophilic granuloma, Hand-Schüller-Christian disease and Letterer-Siwe disease are classified as histiocytosis X. The simultaneous appearance of Langerhans cell granules and Charcot-Leyden crystals is specific in histiocytosis X.


Analysis of atropine action at the frog neutromuscular junction. 1. Atropine action on the end-plate currents (e.p.c.s) has been analysed at the macroscopic and elementary levels. 2. The shortening effect of atropine on the e.p.c. and m.e.p.c. level can be fully explained by a reduction of the life time of the elementary current: this effect is markedly increased at more hyperpolarized membrane potentials and at higher concentrations of atropine. 3. It is therefore suggested that atropine binds to the open acetylcholine-receptor complex, leading to a state with a null conductance. According to this model, the forward rate constant of atropine binding could be calculated and was of the order of 10(7) M-1 S-1 AT -90 MV and 20-22 degrees C. 4. Although the conductance at the peak of the e.p.c. is reduced by atropine and becomes voltage sensitive, the elementary conductance is affected neither by voltage nor by atropine. 5. The exclusive binding of atropine to the activated ACh-receptor complex, as proposed above, does not appear to explain this phenomenon. Another binding occurring before the channel is open with a dissociation constant of 60 micrometer could account for this effect.


The force-velocity relationship in vertebrate muscle fibres at varied tonicity of the extracellular medium. 1. The relationship between active force and velocity of shortening was studied during tetanic contraction of isolated semitendinosus muscle fibres of the frog (0.5-2.0 degrees C). Measurements were carried out with the fibre immersed in isotonic (1.00R) Ringer solution and in solutions that were made hypotonic by reduction of NaCl (osmolality 0.62 and 0.81 of normal Ringer) and hypertonic by addition of sucrose (osmolality 1.22 and 1.44 of normal Ringer).2. The force-velocity relation was hyperbolic at loads lower than 80% of measured isometric force (P(0)) but exhibited a reversed curvature between 0.8P(0) and P(0). The maximum velocity of shortening was determined in two different ways: (i) by extrapolation to zero load from force-velocity data truncated at 0.8P(0) (computer fitting of hyperbola, leastsquares method) and (ii) by recording the time required to take up the slack of the fibre after a quick release during tetanus.3. Isometric force and maximum speed of shortening both changed inversely with the tonicity of the extracellular medium. Immersion of the fibre in 0.81R hypotonic solution caused active tension and shortening velocity to increase by 10 +/- 1% (mean +/- S.E. of mean, n = 14) and 12 +/- 1%, respectively. Conversely, force and shortening velocity decreased by 12 +/- 1% (n = 13) and 22 +/- 2% when normal Ringer was replaced by 1.22R hypertonic solution. These changes doubled when the tonicity was altered from normal Ringer to 0.62R and 1.44R, respectively.4. Changes in fibre cross-sectional area equivalent to those obtained in the 0.81R and 1.22R solutions (+11% and -13%, respectively) were produced by varying the sarcomere length within the range 2.0-2.5 mum in the normal Ringer solution. Maximum velocity of shortening remained very nearly constant under these conditions, indicating that the shortening velocity, like the isometric force, is not critically dependent on changes in myofilament lattice width over the range considered.5. The results support the view that both shortening velocity and active force are modulated by changes of the intracellular ionic strength above and below the level that normally exists in the intact muscle fibre.


The binding of labelled saxitoxin to the sodium channels in normal and denervated mammalian muscle, and in amphibian muscle. 1. The binding of [3H]saxitoxin to innervated and denervated rat diaphragm muscle, and to normal frog muscle, has been measured. 2. A saturable component of saxitoxin binding, which was inhibited by tetrodotoxin, was detected in all preparations, as well as a component of non-saturable binding. The values for the maximum saturable capacity, M, and the equilibrium binding constant, K, for normal rat diaphragm muscle were: M = 24-4 f-mole.mg wet-1, and K = 3 -8 NM. 3. Denervation of rat diaphragm muscle reduced the maximum binding capacity per unit weight to 16-5 f-mole.mg-1. The value of K remained virtually unchanged at 4-2 nM. 4. It is suggested that the decrease in density per unit weight does not reflect any change in the density of sodium channels per unit area of membrane. 5. Two varieties of the same species of frog, Rana pipiens, were examined. In one variety (Southern) the value of M was 25-6 f-mole.mg-1 and the value of K was 4-3 nM. In the Northern variety the maximum binding capacity was less, M being 14-6 f-mole.mg-1; the value of K was 3-8 nM.


Tension responses to sudden length change in stimulated frog muscle fibres near slack length. 1. Apparatus for applying a step change of length to an isolated muscle fibre is described. The step was complete in about 0.2 ms.2. Effects of tendon compliance were eliminated by using a spot-follower device and by gripping the tendons with metal clips close to the fibre ends.3. The natural frequency of the force transducer was above 10 kHz.4. Steps of various amplitudes and in either direction were applied to isolated muscle fibres about 6 mm long from the anterior tibial muscle of Rana temporaria during tetanic stimulation. Initial sarcomere length was 2.0-2.2 mum, and temperature was 0-3 degrees C.5. The tension response to a step could be divided into four phases. The initial response was an apparently elastic change during the step itself (phase 1). After the step was completed there was a rapid partial recovery towards the original tension (phase 2, lasting 2-5 ms), followed by a slowing or reversal of recovery (phase 3, 10-50 ms), and finally a much slower return to the original tension (phase 4). Most of this paper is concerned with phases 1 and 2.6. The initial tension change (phase 1) occurred synchronously with the applied length change, indicating that the fibres possess a compliance which is almost linear and almost undamped. Its stiffness is such that an instantaneous shortening of about 4 nm per half-sarcomere would bring the tension to zero from its isometric value.7. The absence of detectable damping during phase 1 indicates that the viscosity of a stimulated fibre is substantially less than the apparent viscosity of a fibre at rest.8. The instantaneous force-extension curve approached the length axis at a sharp angle and a negative tension appeared at the force transducer when a very large step was applied. These observations suggest that the structures responsible for the stiffness of the fibre remain rigid when they are not under tension.9. During the few milliseconds after the step (phase 2) the tension recovered part of the way toward the level which existed before the step. In shortening steps the time course of this recovery was adequately fitted by the sum of four exponential terms, and was similar in steps of different amplitude but with a time scale shorter the larger the step. In stretches the slow components were relatively larger than in releases.10. The tension level, T(2), approached during phase 2 depended only on the total amplitude of the step and not on the time course of the length change, provided it was complete in 1-2 ms. The extreme tension reached during a step could thus vary widely without detectable change in T(2).11. With stretches and releases of up to about 3 nm per half-sarcomere this early recovery was almost complete, so that the curve of T(2) against step amplitude was nearly horizontal. With larger releases the line curved downwards, reaching zero in a release of about 14 nm per half-sarcomere.12. When the temperature was raised both the developed tension and the stiffness increased, but the relative increase was greater for tension than for stiffness. The amount of instantaneous shortening needed to bring tension to zero was therefore also increased.13. A set of empirical equations is given which describe adequately the first few milliseconds of the tension change in response to any imposed time course of shortening.14. The rapid elasticity and early tension recovery resemble the response of a combination of two elastic components and one viscous component. Reasons are given for preferring an interpretation in terms of an undamped compliance in series with a damped compliance (Voigt element) rather than an undamped elasticity in parallel with a series combination of viscous and elastic components (Maxwell element).15. The rapid compliance does not correspond to the ;series elastic component' of two-component theories of muscle contraction.


Voltage-clamp experiments on frog single skeletal muscle fibres: evidence for a tubular sodium current. 1. A late inward current following the initial sodium current is sometimes observed on frog single skeletal muscle fibres studied in voltage-clamp conditions by the double sucrose-gap method. 2. The late inward current is time and voltage dependent; it is sensitive to extracellular sodium concentration and inhibited by tetrodotoxin. Therefore, it is very probably carried by sodium ions. 3. It is assumed that the late current reflects a regenerative increase of sodium conductance in the tubular membrane. The main argument favouring this assumption is that the late current is never observed on detubulated fibres. 4. Inactivation studies show a marked difference between the evolution of the two inward currents when the membrane resting potential is increased by previous hyperpolarizations in a high range of values: the late current is decreased while the initial current remains constant. The decrease of the late current is suppressed by tetraethylammonium ions or in a potassium-free medium. On detubulated fibres, the sodium current is never decreased by conditioning hyperpolarizations. 5. It is postulated that a fast transient potassium current, located in the bubular membrane, should develop contemporaneously with the sodium current. 6. The decay of the sodium inactivation is anaylsed on semilogarithmic plot: on normal fibres, it runs in two phases while on detubulated fibres, one phase only is found.


The dependence of the electrical potentials across the membranes of the frog skin upon the concentration of sodium in the mucosal solution. 1. The dependency of the transmembranal potential differences upon the [Na] of the mucosal bathing solution was investigated in frog skin. Semilogarithmic linear correlations were obtained between [na]0 and the transepithelial potential difference (slope: 28-4 +/- 1-8 mV/dec [Na]0), the potential differences across the outer and the inner membrane under open circuit conditions (slope: 33-4 +/- 1-8 and 4-8 +/- 1-5 mV/dec [Na]0, respectively) and the intracellular potential under short circuit conditions (slope: 33-3 +/- 3-4 mV/dec [Na]0). 2. No difference could be observed between skins incubated in Cl- or SO4-Ringer. 3. The results are not in accordance with the Koefoed-Johnsen & Ussing hypothesis. Neither the polarity of the postulated Na electrode at the outer border could be obtained nor the expected slope of the dependency between potential gradient and [Na] of the mucosal bathing solution. 4. Current recirculation through paracellular shunt pathways is suggested to explain the deviation, at least in part. In addition, the I-R-drop resulting from Na entry across the outer border might account for part of the changes of potential gradients. 5. Under all conditions, Na uptake occurs energetically downhill, but permeability changes of the outer border must be postulated to explain the observed dependency of net Na transport upon [Na]0.


[Semi-excitation in nerve and electric nerve model (ENM) (author's transl)]. The "semi-excitation" is an excitation (like phenomenon) found by the authors, first in ENM and then in nerve during observation of the action potential and impedance decrease associated with excitation. The phenomenon was further investigated in relation to the action potential and impedance decrease (for AC of 13 kHz) of the stimulated site on the sciatic nerve and ENM. The nature of the phenomenon was the following. 1. The configuration of the action potential and impedance decrease were almost the same as that of the (complete) excitation, but the durations of the potential change (plateau) and impedance decrease depended on entirely to the duration of stimulating current. 2. Submaximum amplitude of action potential and impedance decrease were graded by the intensity of the stimulating current, but the threshold of this excitation was found. 3. During or after the semi-excitation, absolute or relative refractory period was not found. 4. Both in nerve and ENM, transition from complete excitation to semi-excitation or vice versa was observed. 5. Semi-excitation was found to occurr in the state of reduced membrane potential of the nerve and ENM.


Template-directed reactions of aminonucleosides. We have studied the reactions between adenosine 5'-phosphorimidazolide and 9-(2-amino-2-deoxyxylofuranosyl) adenine (I) or 3'-methylamino-3'-deoxy-adenosine (II), both with and without a poly (U) template. We find that both amino compounds react much more rapidly than does adenosine, in the absence of a template. The rate of reaction is greatly enhanced by a poly (U) template in the case of I, but the enhancement is slight in the case of II.


Cytogenetic study of a new Ph1-positive cell line (NALM-1). The chromosomes of a cell line (NALM-1) derived from the leukocytes of a patient with chronic myelocytic leukemia (CML) were examined with several banding techniques. The modal chromosome number was 46 and the cells contained a Philadelphia chromosome (Ph1), due to the standard translocation of the missing segment of the long arm of chromosome No. 22 onto the distal end of the long arm of chromosome No. 9, i.e., t(9;22) (q34;q11). The Ph1-positive modal cells of the NALM-1 line also had two common marker chromosomes, an extra X-chromosome, and missing chromosomes in groups No. 7, 9, and 15. Immunologic examination of the NALM-1 cells revealed them to have non-T-non-B (null) surface characteristics. An antigen specific for cells of acute leukemia and a human la-like antigen were detected. These facts suggested that the NALM-1 cell line originated from CML cells and maintained the cytogenetic and Immunologic characteristics of such cells.


Cardiovascular surgery in Jehovah's Witnesses. Report of 542 operations without blood transfusion. Jehovah's Witnesses who require operation represent a challenge to the physician because of the patients' refusal to accept blood transfusion. We report a 20-year experience with a consecutive series of 542 Jehovah's Witness patients ranging in age from 1 day to 89 years who underwent operation. Early mortality (within 30 days after operation) was 9.4%. In 362 patients requiring temporary cardiopulmonary bypass, early mortality was 10.7%. Mortality was 13.5% among 126 patients who had single- or double-valve replacement. The only deaths among patients who had aortic valve replacement or repair of a ventricular septal defect occurred in those who had some serious complication before operation. Preoperative or postoperative anemia was a contributing factor in 12 deaths, and loss of blood was the direct cause of three deaths. Cardiovascular operations can be performed safely without blood transfusion.


Changes in working status of patients following coronary bypass surgery. Coronary artery bypass surgery has gained a major role in the management of ischemic heart disease. Relief of symptoms is often the primary goal for these surgical procedures while other effects relating to rehabilitation of such patients have been neglected. To determine change in work status as a result of operation, job status and hours worked before and after surgery were analyzed in 350 patients who had coronary artery bypass grafting procedures. Overall, there was no improvement in return to work or hours worked after surgery. Hours worked before surgery, relief of symptoms, severity of disease, number of bypass grafts placed, and level of education all related substantially to a change in work capacity. It appears that if the potential for enhanced productivity is to be realized, rehabilitative measures must be intensified following coronary artery surgery.


Immunoblastic lymphadenopathy. A report of two cases. Two patients with immunoblastic lymphadenopathy had lymphadenopathy, hepatosplenomegaly, rash, dysproteinemia, constitutional symptoms, and typical morphologic findings or lymph node obliteration by immunoblasts and plasma cells, proliferation of aborizing vessels, and infiltration with amorphous, eosinophilic material. One patient had massive pulmonary infiltrates that responded to steroid therapy early in the course of the disease, but recurred and were found, at autopsy, to represent immunoblastic invasion of the lung. In both cases, a severe peripheral neuropathy developed during the course of the disease. One patient was addicted to dextroamphetamine for 20 years. Response to corticosteroids and cytotoxic therapy was initially rapid and complete, but refractoriness developed as seen in postmortem findings of extensive visceral involvement.


Onset of Prinzmetal's angina two years following sudden death syndrome survival. A patient who was resuscitated after an episode of ventricular fibrillation without myocardial infarction outside the hospital developed angina for the first time two years later. Although the clinical pattern did not fulfill all of the criteria of Prinzmetal's variant angina and was found to correspond with episodes of ventricular tachycardia, we feel that Prinzmetal's angina can be implicated as the mechanism of previos ventricular fibrillation. During the patient's second hospitalization, angiography demonstrated a single, moderately stenotic, right coronary lesion which was bypassed. Immediately postoperatively, the patient developed ventricular fibrillation requiring more than two hours of cardiopulmonary resuscitation. Six months after bypass surgery, she experienced an inferior wall myocardial infarction. She is presently asymptomatic with normal heart size.


[Remote results of the aorto-coronary shunting in chronic forms of ischemic heart disease]. Remote results of the aorto-coronary shunting operation performed in 102 patients with ischemic heart disease are reported. Of this number 75 suffered from severe angina pectoris at rest, while the others had angina pectoris on effort of some or other degree of intensity. An excellent late result was achieved (over periods from 6 months to 5 years) in 33.3, good--in 49.5, satisfactory--in 10.1% of the cases, with the surgery being of no avail in 5.1 and aggravating the patients' condition in 2.0% of the instances. Excellent and good results were obtained in 83% of the cases, viz. disappearance of angina pectoris, improved ECG at rest and on effort. The excellent and good results did not depend on the initial coronary reserve, the degree of the coronaries lesion and on the adequacy of the operation, whereas all the other results (satisfactory, ineffective, deterioration) were seen, as a rule, with an inadequate aorto-coronary shunting in patients with extensive cicatricial changes of the myocardium. A direct relationship between remote results and the extent of the myocardial lesion was established.


T-cell deficiency in immune complex glomerulonephritis. Mice chronically infected with the virus of lymphocytic choriomeningitis (LCM) develop immune complex glomerulonephritis. Others have shown that adoptive immunization of these mice by the i.p. injection of syngeneic immune spleen cells terminates the chronic viral carrier state. The present studies were designed to define the effector cell from the immune spleen responsible for adoptive immunization and to determine the effect of this procedure upon the immune complex nephritis which occurs in LCM carrier mice. The results indicate that the effector cell in adoptive immunization is a T-cell that functions directly as a killer cell when transferred to LCM carrier mice. Sixteen of nineteen adoptively immunized mice examined had less immune complex material deposited in their glomeruli than control unmanipulated litter mates. These data demonstrate that this animal model of immune complex glomerulonephritis is immunodeficient with respect to LCM virus-specific killer T-cells. Transfer of this cell population to the LCM carrier mouse diminishes the animal's viremia and improves its immune complex nephritis. In view of these observations, it is suggested that the rationale for the use of immunosuppressive therapy in spontaneously occurring glomerulonephritis should be carefully reconsidered.


Relationship between dialyzer reuse and the presence of anti-N-like antibodies in chronic hemodialysis patients. One hundred eleven chronic hemodialysis patients from five dialysis units were tested for the presence of antibodies reactive with red blood cell N substance; 77 patients were available for follow-up study after 18 to 24 months. Initially, 18 patients (16%) had serum anti-N-like antibodies. Thirteen of these patients were in a home dialysis program and were reusing hollow fiber dialyzers. The other five had practiced hollow fiber dialyzer reuse in the past. None of 37 patients using coil dialyzers had anti-N-like antibody. On follow-up testing, anti-N-like antibody persisted in all patients restudied except for one who had a successful renal transplant. Anti-N-like antibodies developed in four additional patients: three were reusing hollow fiber dialyzers at the time, but one had not reused dialyzers for 24 months. Statistical analyses indicated that dialyzer reuse, hollow fiber dialyzers, and home dialysis were significantly related to the presence of anti-N-like antibodies. We interpret the clinical and statistical data to indicate that dialyzer reuse is the major clinical factor in the development of anti-N-like antibody. The likely mechanism involves the prolonged exposure of red cells trapped in the dialyzer to formaldehyde used in preparing dialyzers for reuse. No adverse clinical effects of anti-N-like antibodies were evident in our patients, but hemolysis and acute transplant failure have been reported by others.


[Studies on the lymph drainage of the eye. 1. Quantitative registration of the lymph drainage from the orbita of the rabbit with radioactive tracers (author's transl)]. For the investigation on the lymph drainage from the orbita 99mTc-microcolloid, 99mTc-albumin and 198Au-colloid have been injected into the retrobulbar space of 24 rabbits. Measurements of the activity's distribution have been made in vivo with an Anger type camera (pho-Gamma-IV Hp, Searle Nuclear Chicago) and in vitro after section with a sodium iodine crystal well counter (Clinimat-200, Picker). A significant concentration of the activity could be observed for the most part in the equilateral Lymphonoduli cervicales profundi and superficiales and for the less part also in the equilateral Lymphonoduli mandibulares and contralateral Lymphonoduli cervicales profoundi. Furthermore significant activities could be pointed out in the optic nerves as well as in the contralateral retrobulbar space. The data substantiate a lymph drainage from the orbita  as well as a partly reverse "prelymphatic-lymphatic" flow and are of importance for discussions about lymphostatic ophthalmopathy and sympathetic ophthalmitis. For application on man 99mTc-microcolloid (Lymphoscint) shoud be preferred, because with this tracer 1. the lymph drainage can be measured quantitatively quite exactly by extern measurement with the Anger-camera, 2. the allowable radiation dosis for the lens is lying between 0.5-1.0 rad/mCi.


[Fuchs' endothelial dystrophy--the appearance of the corneal endothelium under the electron microscope (author's transl)]. Studying the various stages of Fuchs' endothelial dystrophy with transmission and surface electronmicroscopy makes the dystrophic and degenerative changes in the endothelial and Descemet's membrane visible: dystrophic changes by the formation of tuberosities, whose structure is very similar to that Descemet's membrane, and degenerative changes by reduction in the number of endothelial cells through destruction of the intercellular connections and through cell necrosis. The second stage is the reformation of the structure by fibroblastic cells. The dystrophic cells lead to the formation of a fibrillary tissue between the cells and the tuberosity-laden Descemet's membrane, which is different from the primary structure of this tissue. The origin of these cells is unknown.


[Secondary glaucoma and uveitis: hypertensive uveitis (author's transl)]. The secondary rise of i.o. pressure in uveitis may lead to a true secondary glaucoma or to hypertensive uveitis. The etiology of the endogenous inflammation does not seem to play a role. Pathogenetically the occlusion of the pupil with the formation of iris bombé and the obliteration of the chamber angle by exudate are important factors, while the hypersecretion of aqueous humor plays a minor role. Medical treatment consists in mydriatics and steroids. Surgical treatment depends on the pathogenetic mechanism and consists either in sector iridectomy or a filtering procedure.


[Electro-ophthalmological examination results in fundus heterotopicus (author's transl)]. A single-flash ERG and an EOG were performed in four cases of fundus heterotopicus. While the EOG responses showed a normal light and dark adaptation, the ERG registered a subnormal a- and b-wave, corresponding to the extent of the medial ectasia. Where hemeralopia was present, the ERG showed a monophasic a-wave and a markedly reduced b-wave. That means that in addition to the simple fundus heterotopicus, there is also a form with several clinical characteristics in which a hemeralopia is adaptometrically and electroretinographically demonstrable.


[Immunological studies in a case of congenital nephrotic syndrome with focal extracapillary proliferative glomerulonephritis (author's transl)]. An immunological study in a case of nephrotic syndrome in early infancy with mesangioproliferative glomerulonephritis with focal crescents revealed deficiencies in humoral and cellular immunity. Serum Ig-G antibody levels such as pertussis-agglutinins, tetanus and diophtheria-antitoxing were decreased. A loss of tetanus antibodies in the urine could be observed. The production of tetanus-antibodies after immunogenic stimulation with tetanus-antigen was not diminished. The loss of Ig-A in the urine might have been compensated by a higher synthesis rate, which could also be the cause of the increased serum levels of Ig-M, since a loss of Ig-M in urine was not observed. The diminished serum levels of Ig-G antibodies were considered not only to be the result of loss, but possibly also of higher catabolism, and an insufficient compensation by a higher rate of synthesis. The percentage of T-cells in peripheral blood was found to be low. Stimulation of lymphocytes with PHA was at first normal, but in the last investigations diminished. Special experiments lead to the assumption, that this partial defect of cellular immunity might be the result of lacking or diminution of nutritive plasmafactors.


[Unusual complications in the course of histiocytosis X (author's transl)]. In 3 patients with histiocytosis X the following complications are described: 1. A 16 year old boy suffered from generalized histiocytosis in infancy. He died after acute intracranial hypertension caused by a basilar impression. 2. Recurrent spontaneous pneumothoraces was the first symptom of an initial exclusively pulmonary histiocytosis X. 3. Exophytically growing soft tissue tumors in the late disease state of a progressive histiocytosis X.


[The differentiation of human peripheral blood lymphocytes by immunological methods. III. Results in acute lymphoblastic leukemia (author's transl)]. In 47 patients with acute lymphoblastic leukemia surface markers were evaluated on mononuclear cells of the peripheral blood as well as in some cases on bone marrow lymphocytes. The lymphocytes were characterized by their binding capacity for sheep red blood cells, the demonstration of Fc-receptors, complement receptors as well as surface immunoglobulins. In 6 of 23 untreated patients the blasts bound sheep red blood cells spontaneously (T-ALL), in two of these six cases the lymphoblasts had simultaneously receptors for complement. In a further patients the lymphoblasts had complement- and Fc-receptors. The blasts of 16 of 23 patients were negative in respect to the markers tested (O-ALL). By comparing two groups of patients--one with positive cells, one unreactive--the clinical features differed: the marker positive group showed a predominance of male patients, 5 of 7 patients had a massive mediastinal mass and the remission rate was lower than in the group with positive blasts. 24 patients in remission under maintance treatment had a decreased percentage of rosette forming lymphocytes as well as lymphocytes with surface immunoglobulins and Fc-receptors. There existed some correlation between the percentage of rosette forming lymphocytes and the clinical course: patients with complications had lower percentages of rosette forming lymphocytes than patients with a favourable course.


Differentiation of human lymphocytes by pokeweed mitogen in vitro: studies in normal and in immunodeficient subjects. Immunoglobulin (Ig) synthesis and secretion by peripheral blood lymphocytes activated by pokeweed mitogen (PWM), and by phytohemagglutinin (PHA) was measured in normal individuals and in subjects with primary immunodeficiency. Unstimulated cultures demonstrated stable, low Ig synthesis of all major Ig classes; PWM showed a peak of Ig synthesis at day 5; PHA cultures demonstrated a late peak of ig production at day 9. Three cases of common variable immunodeficiency showed different patterns of data when the percentage of B cells in blood and Ig production in vitro were used as parameters. Two persons with immunodeficiency and thymoma showed decreased Ig production in vitro, and had suppressor cells capable of blocking Ig production by normal lymphocytes in co-cultivation experments.


T & B lymphocytes in the spectrum of leprosy. The percentage of T & B lymphocytes were estimated in 52 leprosy patients by 'E' and 'EAC' rosette techniques. The mean % values for 'T' lymphocytes were significantly lower in lepromatous group as compared with that of tuberculoid and borderline groups. Also, a significant difference was observed in the mean % values of T & B lymphocytes of the borderline and tuberculoid patients and of the normal control group. These findings were correlated with skin smears and lepromin testing.


Vestibular dysfunction associated with benign paroxysmal vertigo. Benign paroxysmal vertigo (BPV) is a clinical syndrome of vestibular origin although generally no evidence of vestibular dysfunction can be demonstrated with conventional tests. In a review of 1350 consecutive dizzy patients, there were 125 with BPV and of these, 33 underwent a quantitative rotational test of vestibular function. The rotational results showed reduces vestibular system gain for these BPV patients. In addition, they could be subdivided on the basis of a normal or shorter cupular time constant (Tc). Separation of patients into diagnostic categories revealed that those categorized as cupulolithiasis and viral labyrinthitis had a normal Tc range and those categorized as trauma and idiopathic had a short Tc. The reduced gain and short Tc in the latter group suggest hair cell and/or nerve damage since these same changes occur in patients with destructive peripheral vestibular disease.


The lateral approach to acoustic tumors. The surgical experience of a decade of acoustic tumor surgery is discussed. This series involves 106 operations. A plea is made to use the "lateral" approach to the cerebellopontine angle which combines the translabyrinthine and retrosinal routes when necessary. The facial nerve is preserved in 88% of the patients using this approach and the mortality rate is only 1 or 2%. The most common complication is cerebrospinal fluid leakage. The posterior, suboccipital approach in acoustic tumors is condemned. Statistics are presented that make the argument of "hearing preservation" by suboccipital surgery academic. Only 11% of the patients in this series of 106 had "good" hearing preoperatively and all of these had normal hearing in the opposite ear.


Cross-sectional and longitudinal evaluations of an endurance training program. A comparison of the cardiorespiratory responses obtained using both a longitudinal and cross-sectional evaluation of an endurance training program was made in two groups of 60 young male military personnel. Both groups were initially tested (T1) and then retested 6 months later (T2). At T1, Group I was a sample of personnel not participating in a training program while Group II had undergone a 5-month endurance program (2-4 mile run/day). At T2, Groups I and II had been participating in the program for 6 and 11 months, respectively. Testing consisted of sub-maximal and maximal determinations of VO2, VE and heart rate (HR) using an interrupted treadmill test. VO2 max was 11% greater in Group II compared to Group I at T1 and increased 10% in Group I at T2. Similar results were also seen for HR submax, HR max and VE max. These results show that similar values indicative of an improved level of cardiorespiratory fitness can be obtained using either a cross-sectional or longitudinal design when relatively homogeneous groups are studied.


Functional capabilities following coronary bypass surgery. Changes in myocardial oxygen supply were evaluated in 19 patients following coronary by-pass surgery (CBS) for angina pectoris (AP). A symptom limited maximal functional graded exercise test (GXT) was performed before and 2-19 (X 7.5) months following CBS. After surgery the patients were urged to resume full activity and were individually managed in a 10-week "at home" exercise program. Increases were demonstrated for predicted oxygen uptake and work load (p less than .002), and rate pressure product (p less than .02) following CBS, suggesting increased myocardial blood flow. Terminal heart rate showed no significant increase. Of 16 patients exhibiting ST segment depression ( greater than or equal to .1 mV) with AP during GXT before surgery, 10 cases experienced total relief of both of these signs after surgery. A correlation between the pre-operative number of occluded coronary arteries (greater than 50%) and the degree of exercise induced ST segment depression revealed no significant trend. Additionally, no significant relationship was found between post-operative GXT results and the number of by-pass grafts performed on each patient. This study supports the premise that the effectiveness of CBS, in improving functional capacity, can be objectively evaluated by a non-invasive functional GXT.


Electric pain control (EPC) of a painful forearm amputation stump. Surgically and pharmaceutically uncontrollable pain in the forearm stump of a 28-year-old mine explosion victim of the Viet-Nam War, was successfully treated by electric pain control (EPC), which led to his full rehabilitation. The electrodes of the pain-control equipment were built into the prosthetic socket to enable treatment to be continued while the patient was working in his full-time occupation as a storeman. The theory underlying this form of treatment is briefly described and the method of manufacture of the modified prosthetic socket is provided, so that similar application of electric pain control can also be used in other patients.


[Receptors for complement and Fc and T-lymphocyte markers on cells of malignant tumours of man (author's transl)]. 32 malignant solid tumours of various histological types were tested for the presence of Fc receptors, complement receptors and T-lymphocyte markers in single-cell suspensions of fresh tissue. Indicator cells were normal SRBC (E), SRBC coated with rabbit antisheep lg (EA) and EA coated with complement (EAC'). Fluorescent microscopy on tumour cells was done with rabbit anti-human IgG-fraction labeled with FITC. Almost all tumours possessed Fc receptors and/or complement-receptors. The presence of T-lymphocyte marker was less frequently found. It is proposed to establish a "receptor classification" in addition to the histologic classification of the tumours.


Epithelial lesions in low flow states: a unifying concept. Several epithelium-lined organs are highly vulnerable in low flow states, and may lead to lesions such as hemorrhagic enteritis, stress ulcer, renal tubular necrosis, acalculous cholecystitis and "shock lung". A hypothesis is presented which postulated that the redistribution of the subepithelial microcirculatory blood flow and the subsequent damage to the epithelial barrier are the crucial processes in their pathogenesis. Presently available evidence in support of this thesis is summarized.


The mechanism of cobalamin-dependent rearrangements. Adenosylcobalamin-dependent rearrangements are enzyme catalyzed reactions in which a hydrogen atom is transfered from one carbon atom to an adjacent one in exchange for a group X which migrates in the opposite direction. In the hydrogen transfer step, the mechanism of which is reasonably well understood, the cofactor serves as an intermediate hydrogen carrier. The transfer of hydrogen to the cofactor involves homolysis of the carbon-cobalt bond to generate cob(II) alamin and the 5'-deoxyadenos-5'-yl radical, followed by abstraction of a hydrogen atom from the substrate to form 5'-deoxyadenosine and the substrate radical. After migration of group X, the hydrogen atom is returned to the product radical by the reverse of the above reactions to generate the final product and reconstitute the cofactor. In contrast to the transfer of hydrogen, the mechanism of group X migration is poorly understood. Many reactions mechanisms have been proposed on chemical grounds, but there is insufficient biochemical evidence to permit a choice among these propsals. A quantity of negative evidence has accumulated suggesting that group X migration does not involve alkylation of the cobalt of cobalamin by the substrate, but in the absence of firm data supporting an alternative mechanism, even this weak conclusion must be regarded as provisional.


Evidence that UV-inducible error-prone repair is absent in Haemophilus influenzae Rd, with a discussion of the relation to error-prone repair of alkylating-agent damage. Haemophilus influenzae Rd and its derivatives are mutated either not at all or to only a very small extent by ultraviolet (UV) radiation, X-rays, methyl methanesulfonate, and nitrogen mustard, though they are readily mutated by such agents as N-methyl-N'-nitro-N-nitrosoguanidine, ethyl methanesulfonate, and nitrosocarbaryl. In these respects H. influenzae Rd resembles the lexA mutants of Escherichia coli that lack the SOS or reclex UV-inducible error-prone repair system. This similarity is further brought out by the observation that chloramphenicol has little or no effect on post-replication repair after UV irradiation. In E. coli, chloramphenicol has been reported to considerably inhibit post-replication repair in the wild type but not in the lexA mutant. Earlier work has suggested that most or all the mutations induced in H. influenzae by NC result from error-prone repair. Combined treatment with NC and either X-rays or UV shows that the NC error-prone repair system does not produce mutations from the lesions induced by these radiations even while it is producing them from its own lesions. It is concluded that the NC error-prone repair system or systems and the reclex error-prone system are different.


Attempts to induce mutations in Haemophilus influenzae with the base analogues 5-bromodeoxyuridine and 2-aminopurine. Attempts were made to induce mutations in Haemophilus influenzae with the base analogues 5-bromodeoxyuridine and 2-aminopurine. These attempts were unsuccessful. Incorporation studies with BrdUrd showed, in agreement with earlier studies on Escherichia coli, that BrdUrd was discriminated against when dThd was also present but was incorporated to essentially the same extent as dThd when only BrdUrd was present. In this latter case, strands fully substituted with BrdUrd was produced, but survival data suggest that bacteria deriving their DNA by replication on such fully substituted templates were inviable. However, bacteria with about 20% of the thymine substituted with bromouracil were usually viable. No mutations could be detected in the descendants of such bacteria. The reasons for this are discussed and it is concluded that in all probability the replication system in species rarely if every treats incorporated bromouracil as anything except a thymine analogue. The alternative possibility, that the negative results are a consequence of the absence of the reclex (SOS) error-prone repair system in this species, is considered much less likely.


Use of plasma pharmacokinetics to predict and prevent methotrexate toxicity. To correlate the pharmacokinetics and toxicity of methotrexate, we measured the drug's clearance from plasma after 395 high-dose, six-hour infusions given to 78 patients. After 375 infusions, 48 hour methotrexate levels fell within 2 standard deviations of the mean for nontoxic infusions, and myelosuppression did not occur. Methotrexate concentrations exceeded the range for nontoxic patients (mean +/- 2 standard deviations) after 20 infusions. Serious myelosuppression occurred after six of these 20 infusions, including five of 12 infusions associated with 48-hour drug concentrations above 9 X 10(-7) M. In seven patients with 48-hour concentrations above 9 X 10(-7) M, the absence of toxicity could be attributed to subsequent rapid clearance of the drug; four of these patients also received large doses of supplemental leucovorin (50 to 100 mg per square meter every six hours). Determination of methotrexate concentration in plasma thus identified patients at high risk of toxicity, a group that may benefit from supplemental leucovorin rescue.


Intrapetrous epidermoid deforming the pantopaque cisternogram. Pantopaque cisternography is now an established procedure for the diagnosis of mass lesions arising in, or extending into the cerebellopontine angle. Non-filling of the internal auditory canal, even if of normal dimensions, is, with rare exceptions, diagnostic of an intracanalicular acoustic neurinoma. A case is reported of an intrapetrous epidermoid excavating the internal ear, but not widening the meatus, which prevented filling of the canal at cisternography.


Lymphocyte subpopulations in multiple scerlosis: comparison with neuroelectric blocking activity. Lymphocyte subpopulations were determined in patients with confirmed multiple sclerosis using a new technique in which immunoglobulin bearing (lg+) cells are rosetted directly with purified anti-light chain antibody-coated human erythrocytes, while T cells are first sensitized with specific anti-human thymus antisera prepared in b4b4 rabbits, and then indirectly rosetted with purified anti-rabbit light chain allotype antibody-coated human erythrocytes. In 45 percent of the patients, there was a striking decrease in the percentage of T lymphocytes and a reciprocal elevation of lg+ and N cells. In another 20 percent, T cell depression was not accompanied by Ig+ elevation. The remaining 35 percent had normal lymphocyte profiles as did all patients with nondemyelinating neurologic diseases. However, we found no significant correlation between the lymphocyte profiles and prospective clinical evaluation of these patients and no correlation between alterations in lymphocyte profile and the presence of neuroelectric blocking factors in the peripheral blood of multiple sclerosis patients.


Heterozygous alpha-1-antitrypsin deficiency and respiratory function in children. We evaluated 224 children from Rochester, New York, families in which at least one parent was heterozygous for alpha-1-antitrypsin (AAT) deficiency by protease inhibitor (Pi) typing. The childhood population included, of the two major heterozygote Pi types, 75 Pi type MS children and 37 Pi type MZ children. This population was evaluated by means of a standardized respiratory questionnaire, physical examination, and pulmonary function tests. Maximal expiratory flow volume curves were obtained with subjects breathing both air and a helium-oxygen mixture. Total pulmonary resistance by the method of forced oscillations was measured at 3, 5, 7, and 9 cps. No differences in respiratory symptoms or physical findings were seen in Pi type MZ and MS children compared to Pi type MM children. However, when pulmonary function tests were evaluated by a matched-pair analysis designed to minimize other genetic and environmental risk factors, Pi type MZ subjects demonstrated statistically significant differences in forced expiratory flow rates and in increased frequency-dependent characteristics of total pulmonary resistance. These abnormalities are similar to those previously described in adult Pi type MZ subjects, and suggest that physiologic abnormalities associated with AAT deficiency are present early in life.


Double sucrose gap voltage clamp in cardiac muscle. Indirect assessment of voltage control from tension records. In double sucrose gap voltage clamp experimetns on frog atrial bundles the configuration of membrane current and contraction was used to estimate the quality of voltage control. Attention was focused on possible action potential activity along the test segment in response to depolarizing clamps. At low depolarizations large Na+ inward currents were observed while any tension response was missing. Transmembrane potential threshold for generation of a mechanical response was evaluated from conditioned (attenuated) action potentials. The mechanical threshold determined from action potential measurements was only slightly higher than that determined from step-clamp depolarizations. With clamp potentials below the threshold, then, any action potential activity induced by the inward current phase is expected to be rudimentary.


Detection of a contaminant cell culture line by restriction endonuclease cleavage patterns of mitochondrial DNA. A putative HeLa cell culture line was discovered to be contaminated with mouse cells by examination of agarose gel profiles of restriction endonuclease digests of mitochondrial DNA. The contamination was confirmed by karyotypic analysis, and by observation of the mouse satellite band in an analytical buoyant density centrifugation of total cellular DNA. Restriction endonuclease analysis of mitochondrial DNA is suggested as a useful method for monitoring the species of cells in culture.


The quantitation of fluorescence by photography. A method based on theory has been developed for the photographic quantitation of fluorescent substances. DNA stained with ethidium in agarose gels is used as an example of an application of this method. In the course of developing this method we have demonstrated that the empirical methods employed by others authors can give rise to large systematic errors. We have also developed an approximate method based on photographic theory, avoiding the use of digital integration which is required by the rigorous method.


[Respiratory function disorders during hepatopathies with immunity involvement]. A respiratory functional test was done systematically on 24 patients with chronic hepatitis and was found normal in only 2 cases. It showed an alteration of the transfer in 14 patients and a syndrom of hyperinflation in 8 others. Despite the histopathological proof it seems that the transfer alteration could be attributed to diffuse interstitial pulmonary lesions. They have already been noted in chronic hepatitis by several authors. The pathogeny of this interstitial involvement probably implied immunity phenomenons including the participation of the antigen Australia or others. As for the hyperinflation syndrome observed in 4 patients in the absence of any bronchopulmonary history, it could reveal an emphysema, which, so far, had not been seen associated to chronic hepatitis.


Specific radiolabeling of a cell surface receptor for epidermal growth factor. A photoreactive derivative of epidermal growth factor (EGF) has been used to identify and specifically label a membrane receptor for EGF on mouse 3T3 cells. Photoactivable EGF, labeled with 125I, was incubated with 3T3 cells and then photolyzed in situ to generate a nitrene capable of reacting with a wide variety of chemical bonds. Analysis of the system by sodium dodecyl sulfate/polyacrylamide gel electrophoresis revealed, besides the band of EGF, only one other major radioactive band, at a position indicating an apparent molecular weight of 190,000. This band was absent when a nonresponsive and nonbinding variant of 3T3 was used. A direct proportionality between binding activity and crosslinked complex formation was demonstrated using a variety of binding conditions. "Down regulated" cells, in which EGF binding activity was greatly reduced by prolonged incubation with an appropriate concentration of EGF, also had a decrease in covalent complex formation proportional to the decrease in EGF binding activity.


Reinnervation of original synaptic sites on muscle fiber basement membrane after disruption of the muscle cells. Regenerating axons form new synapses precisely at sites of original synapses in denervated skeletal muscle. To determine what role the muscle cell plays in this phenomenon, we studied reinnervation of frog muscle at intervals after crushing the nerve and damaging the muscle fibers. Damaged muscle fibers degenerate and are phagocytized, but their basement membrane persists and acts as a scaffold for regenerating muscle cells. Specializations of the basement membrane serve to mark original synaptic sites after nerve and muscle have degenerated. Regenerating axons enter the region of damage and form functional synapses with regenerating myofibers. The new nerve terminals are found almost exclusively at the original synaptic sites, demonstrating that the integrity of the original postsynaptic cell is not necessary for topographically precise reinnervation of denervated muscle.


Studies on T cells in newborns. Higher reactivity of umbilical cord blood lymphocytes to PHA as measured by whole blood microtechnique. The capacity of human foetal lymphocytes to respond to PHA and to form E-rosettes have been compared with data from adult individuals. For this purpose a microculture system that uses whole blood and avoids the problems of lymphocyte separation, has been developed. Foetal lymphocytes reached optimal stimulation with lower dosis of PHA (31,2 microgram/ml) as compared with adult cells (125-252 microgram/ml). However their quantitative response (measured by 14C-thymidine uptake) was equal in both groups. In addition, peripheral T cells (E-rosetting cells) reached values of 36.47 +/- 9% in newborn and 49.6 +/- 10% in normal adult controls. These results are discussed as to the status and development of cellular inmunity in human foetus.


Craniopharyngiomas in children: neuroradiological evaluation. Craniopharyngiomas constitute one of the commonest tumors in the sellar and parasellar region in children. Most often total surgical removal is attempted. Prior knowledge of the extent of the tumor and adhesions to adjacent neurovascular structures is thus necessary prior to deciding the type of surgical intervention. This paper deals with the usefulness of multiple neuroradiographic studies in evaluating these bening neoplasms, based on nearly 50 cases of craniopharyngiomas.


Phagocytic potential of leukaemic B-lymphocytes. Phagocytosis of small latex particles was demonstrated by light and electron microscopy in a proportion of cells from B-lymphoproliferative disorders including chronic lymphocytic, prolymphocytic and hairy cell leukaemia. It was not observed in a case of T-chronic lymphocytic leukaemia or in normal lymphocytes. A significant correlation was observed between the binding of latex to the lymphocyte surface and the degree of phagocytosis. It is suggested that the phagocytic activity of leukaemic B-lymphocytes is mediated through their Fc receptor and that this property may also be present in normal B-cells. Among mononuclear cells, therefore, phagocytosis can no longer be considered exclusively a property of monocytes.


In vitro cross-reacting antibodies from rabbit antisera to rat brain synaptic membranes and thymocytes. Rabbit anti-rat brain synaptic membrane and anti-rat thymocyte antisera were used in cytotoxicity, immunofluorescence, and absorption assays to define the antigenic relationship between the rat brain and thymocytes, and the localizing properties of antibodies. Both antisera cross-reacted with brain tissue and thymocytes. However, anti-synaptic membrane and anti-thymocyte antisera also contained antibodies specific for neurons and thymocytes, respectively. Immunofluorescence showed that antibodies from anti-thymocyte antiserum reacted with antigenic determinants situated on the surface membrane of brain cells.


Direct activation of guinea pig macrophages by a B-cell mitogen. Since polyclonal B-cell activators (PBA) require macrophages to induce modifications in the lymphocyte in vitro primary response to thymus-dependent antigens, we have investigated whether PBA act directly on macrophages. [14C]glucosamine uptake by guinea pig peritoneal adherent cells after stimulation with purified protein derivative (PPD), lipopolysaccharide (LPS), and dextran sulfate (DxS) was tested. PPD produced an increased [14C]glucosamine uptake, whereas LPS and DxS did not. According to our experiments, (a) PPD does not require the presence of lymphocytes to stimulate macrophages; furthermore, when lymphocytes were present in a concentration higher than 5%, a suppressor effect in the glucosamine uptake was found, and (b) there was no significant difference between the findings when peritoneal adherent cells were cultured in normal medium and in supernatant of lymphocyte cultures stimulated with PPD.


Agammaglobulinemia with arthritis and celiac disease developing after infectious mononucleosis. Follow-up study of a case. A previously healthy 2 1/2-year-old boy from a healthy family developed agammaglobulinemia with arthritis 3 months after infectious mononucleosis (IM). The response of serum immunoglobulins at the initial stage of IM was typical, with greatly elevated IgM and a positive IM-specific heterophil antibody test. A secondary celiac disease was diagnosed one year after IM. Considering the serum immunoglobulin levels and the normal half-lives of IgM and IgG, it seems very probable that the synthesis of immunoglobulins ceased about one month after the onset of IM. The role of suppresor T-cells in the development of acquired secondary agammaglobulinemia is discussed.


Cell proliferation in rheumatoid synovial tissue. Rheumatoid synovial tissue cell proliferation was studied by incubating fresh synovial tissue samples with [3H]-thymidine in vitro. The samples were collected at operation from 7 rheumatoid patients. It was found that 0.9-4.9% of the total cell population, excluding the synovial lining cells, became labelled.


Improvement of gait following functional electrical stimulation. I. Investigations on changes in voluntary strength and proprioceptive reflexes. Patients with central spastic paresis and equipped with peroneal stimulators sometimes experience an improvement in their gait, even when the stimulator has been switched off. The object of the present investigation was to reach a better understanding of the mechanisms operating in such cases. 7 patients were investigated on repeated occasions. It was found that some of these patients got a clear increase in isometric strength of foot dorsiflexion following 10 min of peroneal stimulation. In other cases the increase was only slight. There was an inverse relation between the increase and the strength before stimulation. The increase of strength was due, at least partly, to an increased ability to activate the foot dorsiflectors, since there was a simultaneous increase in the EMG from the anterior tibial muscle. Evidence was also obtained suggesting that the increase in strength involved not only foot dorsiflexion but also plantarflexion of the foot end extension of the knee. Following peroneal stimulation there was also a decrease of the achilles reflex and in some cases possibly also the patellar reflex. There was an inverse relation between the decrease in the achilles reflex and its strength before stimulation. It is probable that the changes in voluntary strength and reflex activity reflect conditions which can be of importance in explaining the gait improvement which is sometimes observed in patients equippped with peroneal stimulators.


The effect of electrical stimulation on the experimentally denervated rat muscle. The m. quadriceps of white rats was electrically stimulated after nerve crush and after nerve section. Electrically stimulated muscles showed fewer central nuclei and a greater number of necrotic single fibres. The demonstration of motor endplates could not give a reliable indication of the onset of reinnervation. The various results show the importance of the trophic influence of the nerve and raise the suspicion that electrical stimulation has a retarding effect on the atrophy but also on regeneration of the fibres. After 7 weeks the unstimulated muscles show a greater degree of regeneration than the stimulated ones.


Changes in occupation after coronary arteriography. This study, involving 266 white males evaluates the effect of angiographically determined mild coronary artery impairment upon occupational status. Questionnaires filled out at the time of coronary arteriography and about 13 months later showed that in the interim 65% of the patients stayed in the same occupational status, 31% changed occupational status, and 4% retired. In both younger (less than or equal to 55 years) and older men (greater than 55 years) the proportion of retirees increased after angiography. Of those who remained employed, 19% changed to jobs with higher physical demands while 11% took jobs with lower physical demands. The decision to change the job did not seem to be affected by age or the presence of angina pectoris. Comparison of this patient population with U.S. national average has shown that the presence or awareness of coronary artery disease is associated with measured increase in occupational mobility, regardless of the severity of the disease. Our findings suggest that changes in occupational status after determination of mild coronary artery disease are motivated by psychological rather than physiological factors.


Gas flow in relation to blood flow in oxygenators. An evaluation of the new Shiley bubble oxygenator. A new bubble oxygenator, designed by the Shiley Laboratories, which utilizes a low gas flow for oxygenation, was first tested in dogs and then in 100 clinical cases. For perfusion in adult patients, a gas flow as low as 0.6 litre oxygen per 1 litre blood has given adequate oxygenation and removal of carbon dioxide with no untoward effect on the blood, as compared with other oxygenators, including the membrane type.


[Recurrent massive hemorrhage of the distal part of the digestive tract due to hemorrhagic hereditary telangiectasia]. Massive gastrointestinal bleeding is known to occur in patients presenting the hemorrhagic hereditary telangiectasis condition known as Rendu-Osler-Weber (ROW) disease. Bleeding is most frequent in the fourth decade. Endoscopic coagulation of upper gastrointestinal telangiectatic spots is nowadays possible even in the presence of active bleeding. In the lower digestive tract the naevi are basicallly located on the terminal ileum and the right colon; emergency coloscopic examination is difficult when active bleeding occurs. 10-15% of ROW patients present sudden life-threatening episodes of bleeding. The danger is aggravated through delayed diagnosis as well as difficulty in localizing the hemorrhagic area. Selective angiogram of the three digestive arteries is a safe and efficient method of diagnosing the nature and source of the bleeding. It should be obtained routinely, on an emergency basis, before surgery. Operation includes removal of the involved part of the digestive tract. A specimen angiogram should be obtained during the procedure, to make sure that the bleeding malformation has been removed. Laparotomy without preliminary angiogram is likely to be unsuccessful. An illustrative case is presented.


Aortocoronary bypass: early results of operations in community hospitals of greater New Orleans. All 588 aortocoronary bypass operations performed in the community hospitals of greater New Orleans before 1974 are reported. Hospital mortality averaged 8.7% (4.3% for one bypass; 6.4% for two; 10.3% for three or more; 26% when bypass was combined with another cardiac procedure). Mortality ranged from 5% to 31% among ten surgical groups and from 0 to 20% among seven community hospitals. Men over 60 had excessive mortality (25%). The incidence of major nonfatal complications (myocardial infarction in 12.8% of patients, intraventricular conduction defects in 9.6%, significant arrhythmias in 10.7%, and complications requiring reoperation in 9.5%) also varied with the surgical group, the complexity of the procedure, the patient's age, and the hospital. As determined by these four factors, results of bypass operations in the community hospitals of greater New Orleans ranged from excellent to poor.


Hemophilus endocarditis: new cases, literature review and recommendations for management. Rarely is endocarditis attributed to the species of Hemophilus. Most frequently implicated are H aphrophilus and H parainfluenzae, but H influenzae also is seen. We report six cases of endocarditis due to H aphrophilus or H parainfluenzae and review the literature. Emboli to skin, lungs, kidneys, spleen, brain, and other organs are common complications, and acute glomerulonephritis and meningitis often occur. Ampicillin is the mainstay of antimicrobial therapy for patients whose isolates are sensitive to it, but the duration of antimicrobial therapy necessary for eradication of the infection is not clear. Studies of antimicrobial synergism are warranted in instances of endocarditis caused by ampicilin- or penicillin-resistant strains of Hemophilus, or when patients are allergic to penicillin; in these instances, combination antimicrobial therapy must be given when bactericidal synergism can be demonstrated. Intensive management of complications caused by embolization is crucial to patient survival.


Management of acute traumatic pain by peripheral transcutaneous electrical stimulation. Peripheral transcutaneous electrical stimulation has been used in an attempt to relieve the pain associated with fractured ribs. Sixty-two per cent of the patients considered their pain to be greatly relieved by this electro-analgesic technique and a further 28% gained some relief. Pain was assessed by means of a visual analogue scale. In most of the patients there was a clinical improvement in their condition, the degree of which correlated well with the pain relief. This trial indicated that transcutaneous electro-analgesia is a useful technique for producing analgesia in patients suffering from acute trauma, without some of the side-effects inherent in more conventional forms of therapy.