Biological properties of a Haemophilus influenzae restriction enzyme, Hind I. A type I restriction enzyme from Haemophilus influenzae, Hind I, which requires adenosine 5' -triphosphate and 5-adenosyl methionine, was studied for its activity on transfecting and transforming deoxyribonculeic acid (DNA). The enzyme reduced the size of unmodified bacteriophage S2 DNA from 37 X 10(6) daltons to approximately 10 X 10(6) daltons, but did not affect modified S2 DNA. Unmodified transforming DNA was attacked in vitro by Hind I; however, relatively low levels of inactivation were obtained for single markers, and linked transformants were inactivated as a function of the distance between markers. In contrast, unmodified bacterial DNA was not inactivated in vivo for either single or linked markers by the Hind I restriction system, probably because the segments generated by Hind I were still capable of being integrated in vivo. The lack of preferential inactivation of markers by the enzyme suggests that it makes random breaks in the DNA.


Age and activity effects on rate of bone mineral loss. It has been postulated that the rate of mineral loss in postmenopausal women remains constant with aging and that the decreased activities of daily living associated with aging contribute to mineral loss. These hypothese were examined by measuring the bone mineral content at the midshaft of the radius with the photon absorption technique. The estimated rate of loss was calculated in a cross-sectional study as the regression coefficient of bone mineral content vs. age and in a longitudinal study as the regression coefficient of bone mineral content vs. time. In the cross-sectional study, Group A, which consisted of 264 women aged 50-72 yr, had an estimated rate of loss of -0.0114 +/- 0.0014 (SE) g/cm per yr. Group B, which consisted of 266 women aged 73-96 yr, had an estimated rate of loss of -0.0055 +/- 0.0017 g/cm per yr. In the longitudinal study, Group C consisted of 33 women aged 51-65 yr who were followed for an average of 4.5 yr with a mean number of 16 visits per subject; they were found to have a mean rate of loss of -0.00990 +/- 0.00107 g/cm per yr. Group D consisted of 38 women aged 70-91 yr who were followed for an average of 3.8 yr with a mean number of 31 visits per subjects; they were found to have a mean rate of loss of -0.00020 +/- 0.00236 g/cm per yr. The estimated and directly measured rates of loss were more rapid in the younger groups than in the older groups (A vs. B, P less than 0.001; C vs. D, P less than 0.001). These data demonstrate that the mean rate of mineral loss is not constant with aging and that in elderly subjects it is significantly slower than that of the earlier postmenopausal years. Since the elderly women were the less active, these findings suggest that factors other than decreased physical activity are more important in determining the rates of mineral loss.


Use of a problem-oriented dental record in undergraduate dental education. This paper describes how the concepts and methods of problem-orientation have been applied in the clinical education of dental students. It makes the point that the problem-oriented dental record is only one part of a complete system which has five interrelated components. The problem-oriented dental record is discussed in detail, and the paper includes examples of all four sections of the record. All sections of the record are interrelated, and the steps in identifying and resolving problems follow a logical sequence: (1) collect the data base, (2) write the cue list, (3) write the problem list, (4) write the initial plan, (5) write the final plan, and (6) record progress. The problem-oriented dental record is seen as a tool for patient care, audit, and education. The main advantage lies in the way information is organized and recorded. The record is highly communicative and permits everyone involved an opportunity to relate the data collected to the problem list and to the decisions made regarding how problems will be resolved. The record and the systems were borrowed from Weed and are being adapted for use in dentistry. Although still in the trial stages, this system has proven to have many more advantages than other record-keeping systems and is invaluable as a tool for education. In the future we plan to use computers to help store and retrieve data from these records. We anticipate that a dental education built on this record will have a lasting impact on the graduates' approach to patient care. This should result in a graduate who is conversant with medical colleagues and systematic in the collection of data, listing of problems, and resolution of patient problems. Finally, we believe that the problem-oriented dental record is also a good tool to help the dental graduate become more empathetic with patients and more aware of their total health needs. The record provides a mechanism for recording physical and emotional information as well as the oral status.


[Pathogenic action and sero-immunologic phenomena of Trichomonas vaginalis]. Trichomonas vaginalis is a frequent parasite of the urogenital system of the human been. The pathological manifestations are different with each strain. These variations of the pathogenic action of the parasite are found in the study of the experimental pathogenic action of the animal and in vitro on cell culture. The sero-immunological phenomens which lead to the presence of the parasite are not completely known. Some technics was proposed for the diagnosis. Their practical utilisation is not completely related to the heterogenicity of the antigenic structure of the different strains of Trichomonas vaginalis.


[Consumption coagulopathies and severe postpartum hemorrhage]. 15 patients were referred to the Intensive Care Unit immediately after delivery for severe post-partum haemorrhage. In 7 cases complications followed retroplacental haemotoma formation and in the other 8 cases they were due to secondary haemorrhages from the placental site and/or from tears of the cervix or vagina. It is a clinical picture that is associated with a state of shock, with continuing massive bleeding associated with failure of the blood to clot and which persists and becomes worse with the transfusion of stored blood and of the coagulation factors. In all these cases severe coagulation pathology due to consumption defects is revealed. We must point out that tears of the cervix and vagina which have been neglected and failure to compensate for the original blood loss are among the aetiological factors. Treatment with Heparin giving a dose of 1 to 3 mg per kg of body weight per 24 hours and controlled by a strict biological check together with symptomatic resuscitation from the state of shock and from the coagulation factors is successful (9 cases) if it is undertaken early on. A fatal outcome, whether it is due to a persistent state of shock (1 case) or to visceral complications of diffuse intravascular coagulation (C.I.V.D.) (5 cases) can be found where treatment is delayed and when insufficient blood is replaced and when antifibrinolytics are prescribed. A preventive therapeutic plan as well as a curative one can be drawn up for these cases of obstetrical drama.


Cutaneous basophil hypersensitivity uncovered in the cell transfer of classical tuberculin hypersensitivity. Cellular transfer of cutaneous basophil hypersensitivity (CBH) was studied. Guinea pigs immunized for CBH with incomplete Freund's adjuvant (IFA) provided cells which could transfer delayed and basophil-rich reactions in skin tests of recipients. Guinea pigs immunized with complete classical tuberculin-type delayed hypersensitivity reactions (DH), which are characteristically devoid of basophils. However, recipients of cells from donors with DH, surprisingly, were found to have delayed skin reactions containing large basophil infiltrates which were lacking in the donors. Thus, recipients of classical cell transfers of tuberculin-type DH had delayed reactions which resembled CBH. Control experiments verified that the cell transfer of CBH from donors with DH was due to passive transfer with live cells and not transfer of contaminating humoral factors or active sensitization of recipients. It was concluded that cutaneous basophil responses were suppressed in CFA-immunized donors and expressed in cell transfer recipients. Cells from donors immunized with CFA were enriched for nonadherent and nonimmunoglobulin-bearing lymphocytes by passage through nylon wool columns, and these cells transferred conjugate specific CBH reactions. It was concluded that cells mediating these transfers were probably T cells. The finding of basophils in cell transfers of DH and a variety of other findings suggesting complex regulation of basophil numbers in tissue lead to the conclusion that the term CBH be used to simply describe a basophil-containing skin reaction.


Thymic function in NZB mice. II. Regulatory influence of a circulating thymic factor on antibody production against polyvinylpyrrolidone in NZB mice. Administration of a circulating thymic factor isolated from normal pig blood prevented the development of the exaggerated production of anti-polyvinyl pyrrolidone (PVP) antibody in young NZB mice. However, treatment was ineffective if initiated after the 4th week of life at a time when endogenous serum thymic factor (TF) normally disappears in these mice. These data suggest that circulating TF is necessary for the survival of short-lived suppressor T cells normally implicated in the regulation of the production of antibodies against PVP, a thymus-independent antigen. In older NZB mice, TF treatment increased paradoxically anti-PVP antibody production, which suggests that "amplifier" T cell activity could also be under TF influence.


Influence of H-2 on the ontogenesis of a spleen cell population which lacks C3 receptors and Thy-1 antigen. During ontogeny there are marked differences in the cellularity of the spleen of mice from different inbred strains. Data from segregation analysis demonstrate that one of the genes involved in the control of the size of the spleen at 15 days lies in the H-2 region. In contrast to previous reports, we find that the cell population under H-2 influence lacks complement receptors.


Tolerance and contact sensitivity to DNFB in mice. VI. Inhibition of afferent sensitivity by suppressor T cells in adoptive tolerance. Tolerance in contact sensitivity to DNFB can be adoptively transferred to normal mice with lymph node cells from tolerant donors. This tolerance is antigen specific and is mediated by T cells, i.e., "suppressor" T cells. Experiments were carried out to investigate the mechanism(s) by which the suppressor T cells induce tolerance to DNFB contact sensitivity. The suppressor cells were effective only if they were present during the early stages of the afferent limb of sensitization. As measured by DNA synthesis, cell proliferation in the draining lymph nodes of recipients of suppressor cells was found to be significantly less than in control animals indicating that the suppressor cells acted, at least in part, by limiting or inhibiting DNFB-induced cell proliferation. This inhibition was shown to be antigen specific since the DNFB suppressor cells did not inhibit cell proliferation induced by oxazolone, an unrelated contact sensitizer. The ability to DNFB tolerant cells to block  afferent sensitization pathways differs from the mechanism of tolerance to picryl chloride, reported by others, where efferent pathways are blocked.


Immune response of mice to the Thy-1.1 antigen: intra-H-2 mapping of the complementary Ir-Thy-1 loci. The primary immune response to the Thy-1.1 antigen was measured by a plaque assay that detected cells producing antibodies lytic for AKR thymocytes. F1 hybrids which were b/d heterozygous at the entire H-2 complex or at only some of its regions were studied. All F1 hybrids which were b/d heterozygotes at the K, IA and possibly IB regions displayed a complementary effect of the alleles at the Ir-Thy-1A and Ir-Thy-1B loci. The complementary effect was indicated by the significantly higher responses of the F1 hybrids than the responses of their parental strains. No complementary effect could be demonstrated in F1 hybrids which were b/d heterozygous at the IB, IC, S and D regions, but not at the K and IA regions. The results excluded participation of the IC, S and D regions in the phenomenon of the complementation. Furthermore, one of the Ir-Thy-1 loci could be mapped to the left of the IB subregion, most likely in the IA subregion of the H-2 complex. The second Ir-Thy-1 locus could be mapped to the left of the IC subregion in either the IA or IB subregion.


An appraisal of Fc receptors on human peripheral blood B and L lymphocytes. Human circulating lymphocytes with easily detectable surface immunoglobulin have been divided into two populations, B cells and L cells. This second population lacks membrane-incorporated Ig, but has a receptor for membrane-labile cytophilic IgG. In this study purified B and L lymphocytes were examined for Fc receptors that bind aggregated IgG and IgG complexed to erythrocytes. Purified lymphocyte populations were prepared by nylon columns and by negative selection with rosetting techniques. L lymphocytes bound aggregated guinea pig and human IgG, and formed rosettes with human erythrocytes sensitized with Ripley IgG (EA). Treatment of L lymphocytes with trypsin had no effect on the receptors for IgG. B lymphocytes did not bind EA and attachment of aggregated IgG was variable; up to one-third of these cells fixed aggregated human IgG to the cell membrane. Trypsin treatment abolished binding of Agg-IgG to B cells in sharp contrast to its effect on L cells. Furthermore, double-label immunofluorescence studies showed that cells with both membrane-incorporated Ig and receptors for aggregated guinea pig IgG were rare. These studies indicate that human peripheral blood B lymphocytes lack a high affinity, trypsin-resistant Fc receptor that is present on L lymphocytes.


Replication of dengue-2 virus in cultured human lymphoblastoid cells and subpopulations of human peripheral leukocytes. We studied the susceptibility of four human lymphoblastoid cell lines (HCL) and of subpopulations of circulating peripheral human leukocytes to dengue-2 virus infection. HCL with B cell characteristics (Raji, Wil 2WT, 8866), B-type peripheral lymphocytes, and macrophages were productively infected by dengue-2 virus. In contrast, an HCL with T cell characteristics (MOLT-4), T type peripheral lymphocytes, and polymorphonuclear (PMN) cells did not become infected and replicate dengue-2 virus. PMN cells did not adsorb dengue-2 virus, suggesting lack of viral receptors. However, T-type cultured lymphoblasts and T-type peripheral lymphocytes adsorbed dengue-2 virus, suggesting that the block in viral replication involves some stage of infection occurring after adsorption. Permissiveness of B-type HCL to dengue-2 virus infection was dependent on the virus seed used but the virus titers obtained among the susceptible HCL varied. HCL infected persistently with dengue-2 virus have been established. Human peripheral lymphocytes inoculated after cultivation for 3 days in complete medium alone or complete medium supplemented with mitogens replicated dengue-2 virus. In contrast, unstimulated peripheral lymphocytes inoculated immediately after isolation adsorbed dengue-2 but did not support its replication. Mitogen-treated and untreated macrophages replicated dengue-2 virus equally well. The efficiency of dengue-2 virus replication by macrophages was higher than that of peripheral lymphocytes but lower than that of HCL.


Site of action of a soluble immune response suppressor (SIRS) produced by concanavalin A-activated spleen cells. The cellular site of action of SIRS, a soluble immune response suppressor released by Con A-activated spleen cells which suppresses antibody responses to heterologous erythrocytes by murine spleen cells in vitro, was investigated. Exposure of spleen cells to SIRS for 2 hr at 37 degrees C or 1 hr at 4 degrees C was sufficient to suppress 5-day antibody responses in vitro. Similar exposure of splenic or peritoneal exudate macrophages to SIRS also suppressed antibody responses by untreated splenic lymphoid cells; exposure of splenic lymphoid cells to SIRS was without effect. SIRS did not act via T cells which might have contaminated the macrophage preparations. SIRS-mediated suppression could be partially overcome by an excess of normal peritoneal exudate macrophages, but not by an excess of T or B cells. These data indicate that the target cell of SIRS activity is the macrophage. The results are discussed in the context of macrophage functions that could be affected by SIRS.


T-cell regulation of antibody responses: demonstration of allotype-specific helper T cells and their specific removal by suppressor T cells. Allotype suppressor T cells (Ts) generated in SJL X BALB/c mice specifically suppress production of antibodies marked with the Ig-1a allotype. The studies presented here show that allotypes Ts suppress by specifically removing helper T cell (Th) activity required to facilitate differentiation and expansion of B cells to Ig-1b antibody-forming cells. We show first that Ts and Th belong to different T-cell subclasses as defined by Ly surface antigens. Ts are Ly2+Lyl- and thus belong to the same subclass as cytotoxic precursor and effector cells; Th are Lyl+Ly2- cells and thus belong to the subclass containing cells which can exert helper functions and initiate delayed hypersensitivity reactions. Placing these cells in these two subclasses shows that Th are different from Ts and suggests that they play different roles in regulating antibody responses. The difference in these roles is defined by the evidence presented here showing that Ts attack Th and regulate the antibody response by specifically regulating the availability of Th activity. We show that in allotype suppressed mice, Ts which suppress Ig-1b antibody production have completely removed the Th activity of helping Ig-1b cells without impairing Th activity which helps other IgB B cells. These findings imply the existence of allotype-specific Th for Ig-1b cells (Ig-1b Th). We directly establish that Ig-1b cells require such help by showing that carrier-primed spleen cells from Iga/Iga congenic hybrids help Ig-1a B cells from hapten-primed Igb/Iga donors but do not help Ig-1b B cells from the same donor in the same adoptive recipient.


Two stages of B-cell memory development with different T-cell requirements. We present evidence here for two stages in B-memory cell development, the first of which is T independent and the second T dependent. For these studies, we use a new type of T-deficient mouse (allotype suppressed) which specifically lacks T-helper activity (Th) for a subset of memory B cells responsible for approximately 10% of the overall IgG antibody response. We have shown elsewhere that these mice (SJL X BALB/c hybrids suppressed for Ig-1b) lack Th capable of helping Ig-1b memory cells, although they have normal Th activity for all other IgG memory B cells. This selective Th deficiency allows study of the effects of T depletion on memory development and avidity maturation of one population of B cells under conditions where the bulk of the immune response in the animal is proceeding normally, thus obviating environmental problems due to secondary effects of T depletion. With this sytem, we show that after a single priming dose of 2,4-dinitrophenyl-keyhole limpet hemocyanin, the memory B-cell pool in suppressed and nonsuppressed donors is indistinguishable with respect to magnitude and avidity of the response for all IgG antibodies produced, including Ig-1b antibody, despite the fact that expression of Ig-1b memory cells is prevented in intact Ig-1b-suppressed mice by the absence of Th capbale of cooperating with these memory cells. We have shown elsewhere that virtually all of the Ig-1b memory is carried by Ig-1b bearing cells. In contrast with the lack of suppressor T-cell effect on initial Ig-1b memory cell development, our data show that continued Ig-1b memory development is selectively impaired in suppressed mice. When primed mice are boosted repeatedly with the priming antigen, the average avidity of most of the IgG memory cells increases over 100-fold while there is no avidity increase in the Ig-1b component. To explain these data, we suggest that the development of high avidity memory occurs in two stages. The first stage, which occurs as a result of primary antigenic exposure, is the creation of a pool of IgG-bearing memory cells with a relatively low average avidity for the antigen. The appearance of these first stage memory cells does not require help from (post-thymic) Th, although Th are required for the expression of these memory cells (antibody production). The second stage of B-memory development requires both further antigenic stimulation and B-memory cell interaction with competent Th. This is a continuing process in which the number of memory cells in the pool remains relatively constant but the average avidity of these cells increases with continued antigenic exposure.


Murine B-cell subpopulations responsive to T-dependent and T-independent antigens. Strain A/J mice made secondary indirect plaque-forming cell (PFC) responses to azobenzenearsonate (ABA) conjugates of giant keyhole limpet hemocyanin (KLH), a thymic-dependent antigen, but not to conjugates of Ficoll, a T-independent antigen. ABA-Ficoll was also unable to elicit a response in animals primed with ABA-KLH, which have an expanded anti-ABA memory cell pool. On the other hand, ABA-Ficoll rendered mice unresponsive to ABA-KLH when administered before priming or boosting with the T-dependent immunogen. Hence, the T-independent antigen was able to tolerize but unable to trigger B-memory cells responsive to the T-dependent antigen. A/J mice immunized with dinitrophenyl conjugates of Ficoll or bovine IgG (BGG) made vigorous IgM and IgG PFC responses. PFC responses to ABA-KLH and 2,4-dinitrophenyl (DNP)-BGG were abrogated by depleting mice of C3 with cobra venom factor, whereas the IgM and IgG PFC responses to DNP-Ficoll were unaffected. B lymphocytes were fractionated on the basis of receptors for C3 and the subpopulations were assayed for in vitro PFC responses to DNP-Ficoll. Very little response was obtained from complement receptor lymphocyte [CRL(+)] B cells, whereas CRL(-) cells were more responsive than unfractionated B cells. Both populations responded to a polyclonal B-cell mitogen (lipopolysaccharide). On the other hand, the in vitro PFC response to a T-dependent antigen (sheep erythrocytes) correlated with the presence of CRL(+) B cells in the cultures. However, a minor component of this response, sensitive to anti-Thy-1 serum, was made by CRL(-) B cells, indicating the existence of subpopulations of T-dependent B cells with different signalling requirements. The results suggest that most B cells responsive to T-dependent antigens possess receptors for C3 and that C3 plays an obligatory role in the response of these cells. A distinct subpopulation of B cells which lack C3 receptors respond to T-independent antigens. The precursors of PFC for the ABA epitope reside largely or exclusively in the CRL(+) compartment in A/J mice, whereas precursors for the DNP determinant are found in both compartments.


Heterogeneity of murine regulatory T cells. I. Subpopulations of amplifier and suppressor T cells. Immunization of C3H/HeJ mice with 4 X 10(9) SRBC yields a whole splenic T-cell population which can, upon transfer, specifically suppress recipient direct and indirect plaque-forming cells (PFC) responses to sheep erythrocytes (SRBC). Discontinuous bovine serum albumin density gradient fractionation of these T cells demonstrated a population of low density T cells which augmented and a population of high density T cells which suppressed recipient responses irrespective of the number of T cells transferred. Moreover, infusion of admixtures of low and high density cells resulted in intermediate regulatory functions which could be predicted by knowing the regulatory capacity of each population alone. In addition to heterogeneity existing among regulatory T cells as regards amplification and suppression, it appeared that heterogeneity existed within the suppressor T population. Thus, T cells capable of inhibiting direct PFC could be distinguished from those suppressing indirect PFC by their differential localization in peripheral lymphoid tissue, differences in the dissipation of suppressive influences during incubation at 37 degrees C, and by differences in the possible requirement for adherent cell populations. While the relative frequency of both low density amplifier and high density suppressor cells increased with the dose of SRBC used for their induction, it appeared that suppressor cells might be generated in response to feedback signals from amplifier cells. These studies indicate that further delineation of heterogeneity existing within suppressor populations may be helpful in defining mechanisms required for the induction and manifestation of suppressive regulatory forces.


Expression of Ia antigens on hapten-specific B cells. I. Delineation of B-cell subpopulations. The nonimmune adult spleen contains at least two B-cell subpopulations. The majority of primary B cells express cell surface Ia determinants and have the capacity to give rise to IgG antibody-producing clones after T-cell dependent antigenic stimulation. There is also a small subpopulation of primary B cells which are, by definition, Ia negative, since their activity is not eliminated by negative selection with anti-Ia serum and complement. The Ia-negative B cells give rise to clones that produce only IgM antibody. These B-cell subsets may form a continuum in B-cell maturation, or they may exist as discrete B-cell lineages. Since the cellular expression of Ia antigens appears to correlate with the ability of the B cell to generate IgG-producing clones, it is speculated that Ia molecules may have a role in the IgM to IgG B-cell switch mechanism.


T-Lymphocyte precursors. I. Synergy between precursor and mature T lymphocytes in the response to concanavalin A. When mouse bone marrow cells are mixed with cortisol-resistant thymocytes and stimulated in vitro with concanavalin A, the mitogenic response observed is much greater than additive, that is, it is synergistic. Between 94 and 96% of responding cells could be identified as T cells (Thy-1 positive) and of these, 79-100% derived from the cortisol-resistant thymocyte population, not from the bone marrow. Purified macrophages could not replace bone marrow; and marrow depleted of mature T or B cells worked as well as normal marrow. Thus, T and B cells and macrophages were ruled out as the synergizing cell of bone marrow. Nude spleen contained 10 times as many precursors of T cells as did nude marrow and was 10 times better at synergy with cortisol-resistant thymocytes. This implication of the pre-T cell as synergizer was supported by the finding that the synergistic activity of marrow was lost on preincubation, but maintained if the preincubation medium contained thymosin or cyclic AMP. Thus, the ability to enhance the response of relatively mature T cells to Con A is a property of pre-T cells. It is anticipated that this property will allow more detailed studies of T-cell precursor development in mice, and possibly in man.


T-cell-deficient mice produce more antihapten antibodies against syngeneic than against allogeneic erythrocyte conjugates. T-cell-deficient mice, either anti-thymocyte serum treated or nude mice, were immunized with hapten (4-hydroxy-3,5-dinitrophenyl acetic acid, NNP) conjugates of syngeneic, allogeneic, or xenogeneic erythrocytes. Immunization with syngeneic conjugates led to a stronger anti-NNP response than immunization with allogeneic or xenogeneic conjugates. A study of congenic mouse strains suggested that a prerequisite for this effect was that immunogenic erythrocytes and responding animals shared H-2-controlled characteristics. F1 hybrid erythrocyte conjugates injected into F1 hybrid mice behaved like other syngeneic erythrocytes. The same erythrocyte conjugates injected into either parental strain induced a weak response indistinguishable from the response to allogeneic erythrocyte conjugates. Parental erythrocyte conjugates injected into F1 mice induced an anti-NNP response that was significantly lower than the response to F1 erythrocyte conjugates but significantly higher than the response to allogeneic conjugates. The response of normal mice to syngeneic erythrocytes was weaker than the response of T-cell-deficient mice, which could have been caused by suppressor T cells. Their response to allogeneic conjugates was higher than the response of T-cell-deficient mice and the response to xenogeneic conjugates higher still. This was probably due to allo- or xenoreactive helper cells.


H-2 compatibility requirement for virus-specific T-cell-mediated cytolysis. Evaluation of the role of H-2I region and non-H-2 genes in regulating immune response. Lymphocytic choriomeningitis virus (LCMV) and ectromelia virus-specific T-cell-mediated cytotoxicity was assayed in various strain combinations using as targets peritoneal macrophages which have been shown to express Ia antigens. Virus-specific cytotoxicity was found only in H-2K- or D-region compatible combinations. I-region compatibility was not necessary nor alone sufficient for lysis. Six different I-region specificities had no obvious effect on the capacity to generate in vivo specific cytotoxicity (expressed in vitro) associated with Dd. Low LCMV-specific cytotoxic activity generated in DBA/2 mice was caused by the non-H-2 genetic background. This trait was inversely related to the infectious virus dose and recessive. Non-H-2 genes, possibly involved in controlling initial spread and multiplication of virus, seem to be, at least in the examples tested, more important in determining virus-specific cytotoxic T-cell activity in spleens than are Ir genes coded in H-2.


Terminal deoxynucleotidyl transferase is found in prothymocytes. Terminal deoxynucleotidyl transferase is an enzyme which has the unique property of polymerizing polydeoxynucleotides onto a primer in the absence of a template (1,2). This enzyme is found both in the thymus and the bone marrow of birds, rodents, and humans (3-7). Whether the marrow cells that contain terminal transferase are related to thymocytes, or are on a separate pathway of differentiation, is not yet known (7,8). To determine the lineage of the murine bone marrow cells that have terminal transferase, we have investigated whether these cells have the antigen Thy-1 induced on the cells by treatment with thymopoietin (9). Thymopoietin is known to induce a set of characteristic T-cell markers including the Thy-1 alloantigen on the surface of a subpopulation of bone marrow cells committed to T-cell differentiation (prothymocytes) (10). Destruction of Thy- 1-positive cells after exposure to thymopoietin allows elimination of a substantial fraction of those bone marrow cells that can repopulate an irradiated thymus (11). We find that such an elimination after induction with the thymic polypeptide removes a substantial amount of terminal transferase from the bone marrow cell population, suggesting that at least one-half of the marrow cells bearing this enzyme are related to those found in the thymus.


Requirement for persistent extracellular antigen in cultures of antigen-binding B lymphocytes. A system was established to assess the requirement for continuous presence of antigen in B-lymphocyte activation to antibody formation. Mouse spleen B lymphocytes, enriched for cells bearing anti-NIP (hapten 4-hydroxy-3-iodo-5-nitrophenylacetic acid) receptors, were pretreated briefly with NIP-POL (polymerized flagellin) antigen, washed, and added in small numbers to microcultures. The behaviour of these cells was compared with that of cells cultured in the continuous presence of antigen. Unfractionated spleen cells were studied under similar conditions. In contrast to unfractionated cells, enriched cells could not be triggered effectively by brief contact with antigen at any concentration tested. Fewer cells were activated, and clone size was smaller after brief contact with antigen than when antigen was present continuously. Furthermore, brief contact at high concentration did not cause tolerance induction.


Induction of an antibody response in cultures of human peripheral blood lymphocytes. A culture system is descirbed which provides adequate conditions for in vitro immunization of humand peripheral blood lymphocytes to heterologous erythrocytes. Making use of this method we could obtain, with a number of different donors, an antibody response which peaked at about day 8 of culture with 30-300 plaque-forming cells (PFC) per 10(6) input lymphocytes. However, in a number of experiments poor or negative results were obtained, even with donors that had previously given good response. This variability in the results was shown not to be due to a too low number of precursor cells present in the blood and could be overcome by treating the cells, before initiation of the culture, with a factor produced by mouse T cells educated to sheep erythrocytes (SRBC). Under these conditions a PFC responce was obtained which peaked at about day 8 and which in some experiments could be as high as 20,000 PFC per 10(6) input lymphocytes. Paralleling the increase in PFC was an increase in cell number. The cells recovered from the treated cultures were at all times more numerous than in the nontreated cultures. The height of both the proliferative and antibody-producing responses varied from experiment to experiment, a higher proliferative response, accompanying a higher PFC response. Although the mechanisms that are at the basis of the antibody response in vitro described in this paper still need to be clarified, this system may become a useful tool in studying the immune response in man.


Helper function of T cells depleted of alloantigen-reactive lymphocytes by filtration through irradiated F1 hybrid recipients. I. Failure to collaborate with allogeneic B cells in a secondary response to sheep erythrocytes measured in vivo. Helper T cells were obtained by injecting heavily irradiated semiallogeneic mice with lymph node cells from H-2-incompatible parental strain mice primed with sheep erythrocytes (SRC) 2 mo before. Thoracic duct lymphocytes collected from the recipents 18-40 h later (nearly all of which were theta-positive and of donor origin) were totally and specifically unresponsive against host-type determinants in mixed-lymphocyte culture. The filtered cells were transferred to irradiated semiallogeneic mice together with SRC and anti-theta-serum-treated (B) cells from SRC-primed syngeneic, semiallogeneic, or allogeneic mice. When antibody-forming cells were measured in the spleen 5-9 days later, effective IgM and IgG collaborative responses were observed with both syngeneic and semiallogeneic B cells but not with allogeneic B cells. No evidence was found that the failure to obtain collaboration with the allogeneic B cells was due to inhibition of the B cells by the T cells or vice versa.


Loss of suppressor T cells in adult NZB/NZW mice. We have investigated suppressor T-cell activity in female NZB/NZW F1 mice using PWM-driven IgM biosynthesis in vitro as an indicator system. In initial we studied we observed that spleen cells from normal mice (BALB/c, C57BL/6), as well as from young (4 wk) and adult (18 wk) NZB/NZW mice, cultured in the presence of PWM synthesize 860 +/- 120 ng IgM/10(6) cells/7 days. However, when Con A (at 2 mug/ml) was added directly to the cultures (along with PWM), cells obtained from adult normal mice and young NZB/NZW mice showed a 94% suppression of IgM synthesis, whereas cells obtained from adult NZB/NZW mice were suppressed significantly less. To analyze these findings we studied the effect of Con A-induced suppressor cells (cells cultured with Con A for 24 h and washed free of Con A) on PWM-driven IgM biosynthesis. Spleen cells obtained from normal mice cultured in the presence of Con A-pulsed cells obtained from normal mice and young NZB/NZW mice showed an 83-88% suppression of PWM-driven IgM synthesis. Similarly, supernates obtained from Con A-pulsed cells of normal mice or of young NZB/NZW mice suppressed PWM-driven IgM synthesis. This suppression by Con A-pulsed cells and their supernates required T cells since T-cell fractions but not B-cell fractions eluted from anti-Fab Sephadex columns mediated suppression of co-cultured normal cells; in addition, Con A-pulsed cells treated with anti-theta and complement do not mediate suppression. These studies of Con A-induced suppressor cell activity in normal mice and young NZB/NZW mice contrast with studies of Con A-induced suppressor cell activity in adult NZB/NZW mice. We found that adult NZB/NZW Con A-pulsed cells and supernates obtained from the Con A-pulse cells had vastly decreased suppressor potential; in this case the Con A-pulse cells and supernatant fluids derived from such cells did not suppress PWM-driven IgM synthesis by normal cells. Finally, whereas spleen cells from young and adult NZB/NZW mice differ in their suppressor cell potential, cells from both sources could respond equally to suppressor signals in that Con A-pulsed normal cells or supernates derived from such cells caused equivalent suppression of PWM-driven IgM synthesis by young and adult NZB/NZW cells. These observations allow us to conclude that NZB/NZW mice lose suppressor T-cell activity as they age.


Selective expression of H-2 (i-region) loci controlling determinants on helper and suppressor T lymphocytes. Data presented here show that locidentify in the I-region of the H-2 gene complex are selectively expressed in different functional T-cell subpopulations. These loci are closely linked (or possibly identical) to loci that control immune responses. They control surface determinants which identify helper and suppressor T lymphocytes. Determinants described here on allotype suppressor T cells (Ts) are found on normal (nonsuppressed) lymphoid cells, but are not found on helper T cells (Th). These determinants are controlled by a locus mapping in the I region of the H-2 complex. In an accompanying publication we show that this locus (Ia-4) marks a new I subregion (I-J) and is expressed only on T cells. Thus Ia-4 determinants idenfity a T-cell subpopulation which includes Ts but not Th. Th also carry identifying surface determinants controlled by loci that map to the H-2 complex, probably within the I region. These determinants are not found on Ts. Data presented also establish that loci in the I region control determinants on Th, but do not conclusively demonstrate that these are the determinants that distinguish Th from Ts. The selective expression of H-2-controlled  determinants on Ts and Th suggests that these determinants are directly involved in immunoregulation.


A new I subregion (I-J) marked by a locus (Ia-4) controlling surface determinants on suppressor T lymphocytes. In an accompanying publication we show that a subpopulation of T lymphocytes, which includes allotype suppressor T cells, selectively expresses I-region determinants. In this report, we show that these determinants are controlled by a new locus, Ia-4. Unlike the classically defined Ia antigens, they are not found on B lymphocytes. Antibody against Ia-4 determinants cannot be detected by conventional dye exclusion cytoxicity assays, suggesting that they are present on a small subpopulation (less than 10%) of peripheral T lymphocytes. The Ia-4 locus marks a new I subregion, provisionally designated I-J. This chromosomal segment is defined by the crossover positions in strains B10.A(5R) (K-end boundary) and B10.HTT (D-end boundary), and maps between the I-B and I-C subregions.


Properties of the antigen-specific suppressive T-cell factor in the regulation of antibody response of the mouse. IV. Special subregion assignment of the gene(s) that codes for the suppressive T-cell factor in the H-2 histocompatibility complex. The locus of the gene that codes for the antigen-specific suppressive T-cell factor was determined to be in a new subregion "I-J" which locates between I-B and I-C subregions in the H-2 histocompatibility complex. This was shown by two different lines of evidence: (a) The absorbing capacity for the suppressive T-cell factor of several alloantisera against restricted I subregions did not correlate with their specificity for previously known Ia molecules which are coded for by genes in I-A and I-C subregions, but was associated with the specificity for the products of genes putatively present between I-B and I-C subregions. By the occurrence of special recombinant strains, i.e. B10.A(5R), B10.A(3R), B10.S(9R), and B10.HTT, which differ with respect to the I-J subregion, we were able to produce alloantisera which distinguish I-J subregion gene products. The absorption studies using these special alloantisera directed to I-J subregion clearly indicated that the suppressive T-cell factor is a product of I-J subregion gene(s), and that the molecule is distinct from known Ia molecules expressed on splenic B cells. (b) Taking advantage of the fact that there is a strict histocompatibility requirement for the effective suppression between the donor and recipient strains of the suppressive T-cell factor, we were able to determine the required identities of the genes in the H-2 complex existing among those present between I-B and I-C. Again, utilizing the T-cell factors obtained from special recombinant strains, i.e. B10.A(4R) and B10.A(5R), we were able to locate the gene that codes for the suppressive T-cell factor reactive only with relevant haplotype strains between I-B and I-C subregions. These results are most reasonably explained by the presence of a new subregion I-J which is specialized in coding for the suppressive T-cell factor as a different molecule from previously known Ia molecules.


H-2 restriction of virus-specific T-cell-mediated effector functions in vivo. II. Adoptive transfer of delayed-type hypersensitivity to murine lymphocytic choriomeningits virus is restriced by the K and D region of H-2. In mice, primary footpad swelling after local infection with lymphocytic choriomeningitis virus (LCMV) and delayed-type hypersensitivity (DTH) adoptively transferred by LCMV immune lymphocytes are T-cell dependent. Nude mice do not develop primary footpad swelling, and T-cell depletion abrogates the capacity to transfer LCMV-specific DTH. Effector T cells involved in eliciting dose-dependent DTH are virus specific in that vaccinia virus-immune lymphocytes could not elicit DTH in LCMV-infected mice. The adoptive transfer of DTH is restricted to H-2K or H-2D compatible donor-recipient combinations. Distinct from the fowl-gamma-globulin DTH model, I-region compatibility is neither necessary nor alone sufficient. Whatever the mechanisms involved in this K- or D-region associated restriction in vivo, it most likely operates at the level of T-cell recognition of "altered self" coded in K or D. T cells associated with the I region (helper T cells and DTH-T cells to fowl-gamma-globulin) are specific for soluble, defined, and inert antigens. T cells associated with the K and D region (T cells cytotoxic in vitro and in vivo for acute LCMV-infected cells, DTH effector T cells, and anti-viral T cells) are specific for infectious, multiplying virus. The fact that T-cell specificity is differentially linked with the I region or with the K and D regions of H-2 may reflect the fundamental biological differences of these antigens. Although it cannot be excluded that separate functional subclasses of T-effector cells could have self-recognizers for different cell surface structures coded in I or K and D, it is more likely that the antigen parameters determine whether T cells are specific for "altered" I or "altered" K- or D-coded structures.


Studies on the mechanism of phagocytosis. II. The interaction of macrophages with anti-immunoglobulin IgG-coated bone marrow-derived lymphocytes. We have examined the effect of the distribution of anti-immunoglobulin IgG molecules on the surface of bone marrow-derived lymphocytes upon the interaction of these cells with macrophages. Lymphocytes which were diffusely coated with antibodies to surface immunoglogulin were ingested by macrophages. Lymphocytes which had the same number of anti-immunoglobulin IgG molecules redistributed to one pole of the surface bound to the macrophages' Fc receptors but were not ingested. These results confirm our previous hypothesis that ingestion of an immunologically coated particle requires the sequential, circumferential binding of specific receptors on the plasma membrane of a phagocytic cell to immunologic ligands distributed over the entire particle surface. Macrophages which had bound capped lymphocytes by the macrophages' Fc receptors removed the immune complex caps from the lymphocyte surface without destroying the lymphocytes. These lymphocytes remained attached to the macrophage surface. The finding that macrophages can phagocytize immune complexes from the surface of a cell without destroying the cell to which these complexes are attached may be important in understanding the effects of antigens and antibodies on cells participating in a humoral immune response, in identifying the mechanisms by which chronic viral infections are established, and in defining the roles of blocking antibodies in tumor immunity.


Secretion of Ia antigens by a subpopulation of T cells which are Ly-1+, Ly-2-, and Ia-. It was found that Ia antignes are rapidly secreted by a subpopulation of splenic T lymphocytes which are nonadherent and which express the surface phenotype Ly-1+, Ly-2-, and Ia-. Secretion of the Ia antigens was a metabolically active process which was inhibited by sodium azide and by Pactamycin, an inhibitor of protein synthesis.


Interferon production in athymic nude mice. Athymic (homozygous nude) mice of a non-inbred stock had relatively little antiviral activity in their serum compared with normal control mice at 4, 6 and 6.5 h after the intraperitoneal injection of Newcastle disease virus. The antiviral activity in the serum had the characteristics of interferon. At 10 h after injection and thereafter, the serum titres were comparable in nude and normal control mice. Exceptional nude mice with thymus-like tissue sometimes produced interferon more or less normally. Transfer of spleen cells from normal donor mice, but not from nude donors, led to increased serum interferon levels in nude recipient mice at 4 h after virus injection.


Changing to problem-oriented methods. Implementation in psychiatric institutions. The so-called problem-oriented methods for organizing and recording clinical information offer many potential benefits to users in psychiatric institutions. Beyond the mechanical aspects of implementation, incorporating a problem-oriented approach into the practices of clinical teams entails conceptual and practical readjustments of considerable magnitude. Based on an 18-month study of eight psychiatric teams with diverse characteristics, the paper discusses rationales and priorities, as well as administrative and educational considerations in the conversion process. Such a process must be studied and understood in setting objectives and channeling resources, if outcomes are to match the expectations.


EEG, 5-HTP-metabolism and acupuncture.

EEG, 5-HTP-metabolism and acupuncture. Correlations between EEG-alterations and administration of 5-HT and 5-HTP in man are described. Age-dependence can be observed. Successful acupuncture produces EEG-alterations especially in younger patients as seen after exogene administration of 5-HT respectively 5-HTP i.v. Since biochemical indications for an increased 5-HT turnover during acupuncture of several acupuncture-points are evident further investigations should be carried out to enable reliable correlations.


Simultaneous determination of eight vitamin D2 isomers by high-speed liquid chromatography. The simultaneous determination of vitamin D2 and its isomers by high-speed liquid chromatography (HSLC) is described. Preferred operating conditions for the resolution and quantitation of vitamin D2 5,6-trans-vitamin D2, ergosterol, lumisterol2, isovitamin D2, isotachysterol2, tachysterol2, and previtamin D2 are as follows: column, "Zorbax" SIL (25 cm x 2.1 mm i.d.); pressure, 100-120 kg/cm2; temperature, ambient; detector, UV 254 nm; mobile phase, 0.15% methanol + 2% ether in pentane (for the simultaneous determination), 10% ether in hexane (for quantitation of the practical sets of isomers), or 55% CHCl3 (distilled) in pentane (for separation of isovitamin D2 and 5,6-trans-vitamin D2); sample size, 1 mul (greater than 1-5 ng); internal standard, p-cresol or alpha-naphthol. High-speed, good resolution, precise and accurate trace analysis, greater analysis flexibility, no necessity for preparing derivatives, and gentle operating conditions are main potential advantages of our method which has proved to be very efficient and surpasses all analytical procedures hitherto proposed.


Simultaneous determination of eight vitamin D2 isomers by proton magnetic resonance spectroscopy. Simultaneous determination of vitamin D2 and its isomers, with the aid of pmr employing a paramagnetic shift reagent Eu (dpm)3, is presented. Thus, a mixture of vitamin D2 isomers is acetylated and its pmr spectra are run with and without the addition of the shift reagent. Relative paramagnetic shift values (gamma rel) of the acetoxyl protons of the acetylated isomers are estimated for solutions in CCl4 against the reference acetoxyl signal of vitamin D2 acetate. Estimated values are as follows: vitamin D2 acetate = 100, 5,6-trans-vitamin D2 acetate = 71 +/- 1, ergosteryl acetate = 86 +/- 1, lumisteryl2 acetate = 123 +/- 3, isovitamin D2 acetate = 82, isotachysteryl2 acetate = 97, tachysteryl2 acetate = 100 (105 +/- 1 in C6D6), and previtamin D2 acetate = 80 +/- 2. On the basis of a dramatic shift of an acetoxyl signal and a constancy of each gamma rel, this method permits immediate identification of the isomers and provides valuable information on their stereochemistry. This technique also facilitates a simultaneous determination of these isomers in a mixture by estimating each signal area.


Antibodies to nucleic acids in congenital immune deficiency states. The presence of antibodies to native DNA, single-stranded DNA, and double-stranded RNA was determined for 37 patients with selective IgA deficiency, 11 patients with Wiskott-Aldrich syndrome, seven patients with common variable agammaglobulinemia, 14 patients with ataxia telangiectasia, six patients with intestinal lymphangiectasia, and one patient with Nezelof syndrome. Of 37 patients with selective IgA deficiency, 11 had antibodies to at least one nucleic acid; six had antibodies to native DNA, seven had antibodies to single-stranded DNA, and four had antibodies to double-stranded RNA. The only other congenital immune deficiency disease studied in which antibodies to nucleic acids were found  was the Wiskott-Aldrich syndrome; in this group three of 11 patients had antibodies to native DNA. Retrospective analysis of our patients with SLE disclosed a 2.6% prevalence of IgA deficiency, a prevalence clearly higher than in the general population. These studies provide further evidence of the association between autoimmunity and abnormalities of IgA production and suggest a relationship between thymic-derived immune regulation and IgA production.


Neonatal hypoxia and pulmonary vasospasm: response to tolazoline. Forty-six neonates with hypoxemia were treated with tolazoline, a pulmonary vasodilator, within the first two days of life. Eight of ten (80%) infants without apparent lung disease responded with a mean increase in PaO2 of 116 torr within one hour of beginning tolazoline infusions. One of the responding infants and two nonresponders died. Thirty-six additional infants with a variety of pulmonary disorders had severe hypoxemia which was refractory to mechanical ventilation. Twenty-one (58%) responded with a mean increase in PaO2 of 130 torr within one hour after beginning tolazoline and 13 (62%) of these survived. Fifteen patients had little or no improvement in PaO2 following tolazoline and only three (20%) of these infants survived. Responders could not be distinguished from nonresponders by clinical or laboratory features prior to therapy with tolazoline. Fourteen infants experienced complications possibly related to tolazoline.


Remarks concerning 102 watersoluble contrast ventriculographies. The Authors report their experience in using water-soluble contrast medium in ventriculographies having carried out 102 ventriculographies partly with Dimer X and partly with "Uromiro 300". Their conclusions are practically similar to those already reported by the many Authors who have written on this subject: 1) facility of examination particularly in urgent cases; 2) excellent neuroradiological definition of the cases; 3) possibility of obtaining a selective ventriculography; 4) indication especially in space-occupying lesions of the posterior cranial contrast medium; 6) possibility of reducing complication rate by carefully carrying out the examination and by avoiding the use of excessive quantities of contrast. In particular the Authors describe some useful methods for obtaining better technical results from the examination and reducing the complication rate.


A voltage-clamp study of the effect of two lidocaine derivatives on the time course of end-plate currents. 1. Voltage-clamped end-plate currents (e.p.c.s) have been studied in the glycerol-treated Rana pipiens sartorius nerve-muscle preparations in normal Ringer solution and in the presence of QX-222 and QX-314. 2. Both QX-222 and QX-314, the trimethyl and triethyl analogues, respectively, of lidocaine, greatly modify end-plate current kinetics. The altered e.p.c.s still show a true reversal potential, which is essentially the same as the reversal potential before drug treatment. The time course of the altered end-plate currents varies with both membrane potential and drug concentration. 3. In the presence of 0-1-1-0 mM QX-222, end-plate currents decay as the sum of three exponentials: I(t) =I1(0)e-k1t+I2(0)e-k2t+I3(0)e-k3t, where the subscipts 1, 2 and 3 refer to the rapidly, intermediately, and slowly decaying components, respectively. Both the amplitudes, Ij(0), and the decay rates, kj, depend upon membrane potential. 4. Hyperpolarization increases the relative size of the first and third components, i.e. I1(0) and I3(0) increase relative to I2(0). Depolarization increases the relative size of the second component. 5. Hyperpolarization causes a decrease in the decay rates k2 and k3 and causes a slight increase in the decay rate k1. Dependence of the three decay rates on membrane potential is well described by: kj=bjeajv. 6. The Q10 of each of the kj is about 3. 7. Raising QX-222 concentration, at any given membrane potential, augments I1(0) and I3(0) at the expense of I2(0). Raising concentration increases k1 and decreases k3; their voltage-dependence is little affected. 8. At all QX-222 concentrations tested the decay rate k2 is nearly the same as the decay rate of a normal e.p.c. recorded at an equivalent holding potential from the same fibre before drug exposure. 9. End-plate currents in the presence of 0-1 mM-QX-314 show a "major" or rapidly decaying phase and a very small, slowly decaying phase or "tail", but no intermediate component. Only the major component is discernible for end-plate currents in 0-5 mM-QX-314. 10. Voltage- and concentration-dependence of the decay rate of the major component in QX-314 is similar to k1 and QX-222. Voltage-dependence of the tails decay rate appears to be similar to k3. It is hypothesized that the second component in QX-222 represents currents of unaltered or normal conductance kinetics, and that the first and third components in QX-222, as well as the major component and tail in QX-314, represent current of "QX-altered conductance kinetics".


Stimulation of the sodium pump by azide and high internal sodium: changes in the number of pumping sites and turnover rate. 1. The effects of 5 mM azide on [3H]ouabain uptake and 22Na efflux were determined. Both glycoside uptake and 22Na efflux were enhanced by azide. 2. Azide stimulated the Na pump in muscles whose pumping sites had been inhibited by ouabain and then transferred to a glycoside-free solution. This stimulation was observed before detecting any recovery of the initial pumping activity. 3. When both the resting and the azide-stimulated 22Na efflux had been blocked by ouabain, an additional exposure to azide, in a ouabain-free solution, had no further effects on 22Na efflux. 4. It is concluded that the increase in Na pumping caused by azide is due in part to an increase in the number of pumping sites. 5. [3H]ouabain binding was measured in muscles with different intracellular alkali cation concentrations. Variations in [Na]i from 15 up to 50 mM did not significantly affect the amount of glycoside bound. A substantial increase in binding occurred when [Na]i reached 70 mM. 6. It is proposed that the increase in Na extrusion that occurs during the recovery of Na loaded muscles mostly results from an increased turnover rate of the pump rather than from an increase in number of pumping sites.


The effects of very low external calcium and sodium concentrations on cardiac contractile strength and calcium-sodium antagonism. 1. The tension levels of the contractures induced in frog atrial trabeculae by reduction of the bathing Na concentration have been investigated over a wide range of [Na+]o and [Ca2+]o, making use of EGTA to buffer [Ca2+]o under 10(-5)M. 2. The relationship between tension and the quotient [Ca2+]o/[Na+]2o is only an apparent one, and holds approximately for Na concentrations as low as 1-5 mM. 3. However, for low Na concentrations tension depends on the absolute values of both [Ca2+]o and [Na+]o. 4. Extracellular Ca is the essential factor in tension production. The results indicate an apparent threshold for tension near 5 X 10(-8) M-[Ca2+]o. 5. It can be shown that, under the present experimental conditions, the Na concentration around the cells must be lower than about 0-1 mM in 'zero' Na+ solutions. 6. It is shown that the over-all process leading to tension production can be explained in terms of a reaction involving two Ca ions. 7. To account for the results, a modification of the reaction scheme put forward by Lüttgau & Niedergerke (1958) is proposed, involving a competitive action between Ca and Na ions at a free receptor site. 8. To account for the scheme, an example of a mechanism involving both internal and external Na and Ca concentrations is considered.


The formation of metarhodospin380 in the retinal rods of the frog. 1. The formation of metarhodopsin380 (metarhodopsin II) was studied in isolated frog retinas exposed to intense flashes of 120 mus duration. 2. A rapid increase in absorbance at 375 nm during the flash was followed by a slower absorbance increase in the subsequent dark period. The slower increase showed virtual completion after 5 ms. 3. The fast absorbance increase during the flash was due to the formation of metarhodopsin478. The rate of this reaction was dependent on the time course of the flash and on the decay rate of lumirhodopsin. 4. Kinetic analysis indicates that three consecutive reactions occur: the light-controlled formation of lumirhodopsin, its first-order decay to metarhodopsin478 and the conversion of metarhodopsin478 into metarhodopsin380. At 21 degrees C, the decay constants were 2 X 10(4) S-1 (lumirhodopsin) and 1 X 10(3) S-1 (metarhodopsin478), respectively.


The influence of drugs on the kinin-forming system in relation to pregnancy and parturition in the rat. The duration of normal gestation and parturition in the rat can be changed by treatment with drugs which alter the equilibrium of the kallikrein-kinin system. The kallikrein inhibitor, aprotinin, when given from Days 19-22 of pregnancy prolongs gestation. Treatment with aprotinin from Days 20-22 of pregnancy prolongs the parturient process, as does a single dose given on the morning of Day 22. Kallikrein, when administered from Days 19-22 of pregnancy, results in a prolongation of gestation and abolishes the pre-parturient behaviour ('labour'). Parturition is prolonged and many fetuses are stillborn. Soya bean trypsin inhibitor when given from Days 19-22 of pregnancy delays and prolongs parturition; maternal haemorrhage occurs during birth and many fetuses are born dead or are abandoned at birth. It is suggested that the kallikrein-kinin system plays a functional role in the normal process of parturition in the rat.


Probable common origin of a hereditary fundus dystrophy (Sorsby's familial pseudoinflammatory macular dystrophy) in an English and Australian family. A genealogical link was established six generations back between a family living in England and Australia, and one of the families reported originally by Sorsby et al (1949) as suffering from autosomal dominant inflammatory macular dystrophy (fundus dystrophy). The onset--in the fifth decade of life--and the progress of the condition, which usually ends in blindness, has been observed in a number of patients and the prodromal development of a colour vision deficiency in some of them confirmed. This defect is fundamentally different from the X-linked colour vision defects and merits further investigation.


X mapping in man: evidence against direct measurable linkage between ocular albinism and deutan colour blindness. A Newfoundland kindred in which ocular albinism and deutan colour blindness are segregating provides strong evidence against the loci for these two X-borne characters being within direct measurable distance of each other.


Classification criteria for systemic lupus erythematosus. Frequency in normal patients. One hundred healthy women were examined prospectively during their annual physical examination for any of the preliminary criteria of systemic lupus erythematosus (SLE). No patient had more than three of the criteria; 50 (50%) had none. The preliminary criteria were effective in excluding normal patients. Three of 19 patients taking oral contraceptives had serum antinuclear antibody (ANA), whereas only one of 81 non-pill users had serum ANA (P less than .05). None of these patients with positive ANA had clinical evidence of SLE.


Heterotransplantation of cell lines derived from human malignant neoplasms into golden hamsters treated with antithymocyte serum. Thirteen cell lines of tissue culture originated from human malignant neoplasms were transplanted into cheek pouches of adult golden hamsters treated with anti-hamster thymocyte serum. In eleven cell lines, transplantation tests with 3.5 X 10(6) to 6.8 X 10(6) cells resulted in neoplastic growth within 4 weeks. In other two cell lines including one from neuroblastoma and another from Burkitt's lymphoma, the inoculum size had to be raised to 11.8 X 10(6) and 12.2 X 10(6) cells to get tumor growth, respectively. The histological appearance of those tumors was consistent with that of the individual primary neoplasms despite of their long in vitro histories. Anaplasia grading of the tumors appeared not to be correlated with the frequency of take.


Stimulation of lymphocytes of patients administered with a trypsin inhibitor, Trasylol (basic pancreatic trypsin inhibitor pharmaceutical), in vitro with BPTI and other several stimulants. Distinguished lymphocytes stimulation was observed in vitro with one out of four patients received Trasylol therapy. The peak response of DNA synthesis was shown to occur at 100 mug/tube of basic pancreatic trypsin inhibitor (BPTI) itself, about 6 days after addition of BPTI. Reactivity (3H-thymidine incorporation) of lymphocytes of patients after the therapy, to specific antigen, ie., purified protein derivative (PPD) or Bordetella pertussis organisms in vitro, was markedly depressed. On the other hand, that to non-specific mitogen, phytohaemagglutinin-p of pokeweed mitogen, was a little reduced, or rather enhanced after the therapy. Conversly, lymphocytes of an originally tuberculin negative patient were lead to be stimulated with PPD in vitro, after skin test with PPD and Trasylol therapy. These various immune responses including drug-induced exanthemas of patients received the therapy, are discussed in this report, in comparison with immune responses of guinea pigs sensitized with BPTI, immune responses of animals in microorganism infections and the contrary immunological effects of Trasylol already reported.


Clinical significance of bradykinin liberation during cardiopulmonary bypass and its prevention by a kallikrein inhibitor. Activation of the kinin system and effects  of Trasylol (Bayer, A.G. Lebukusen, West Germany), a kallikrein inhibitor, were investigated on 52 patients during hemodilutional cardiopulmonary bypass (CPB). Immediately after the start of CPB, neither elevation of bradykinin nor reduction of plasma kininogen (KGN: a precursor of bradykinin) were observed. During CPB, bradykinin level in the blood was markedly elevated, correlating with the significant decrease of kininogen (p less than 0.001). The longer the CPB time, the more marked the reduction of KGN. In the cases requiring over 60 minutes of CPB, the amounts of bradykinin released (4.6-18.0ng/ml) were sufficient to increase capillary permeability as well as peripheral vasodilatation. As shown by the sufnificant increase of hematocrit (p less than 0.005) and the extreme reduction of vascular resistance found at the end of CPB in the prolonged cases. Infusion of Trasylol into the extracorporeal circuit actually prevented the reduction of kininogen and the increase of hematocrit as well as the extreme decrease of vascular resistance in the cases of over 60 minutes CPB. These results clearly point out that Trasylol is beneficial for the prevention of bradykinin liberation and capillary permeability increase and for the maintenance of optimum peripheral vascular tone during CPB. Furthermore, the significance of these findings with regards to complications during and after prolonged CPB was discussed.


[On secondary endoepithelial dystrophy of the cornea due to non-magnetic anterior chamber foreign body (author's transl)]. Secondary endoepithelial dystrophy of the cornea can be uncharacteristic and lead to misdiagnoses. A foreign body in the anterior chamber producing an endothelial dystrophy is often recognized late, or not at all. Clearing the cornea with glycerin facilitates the gonioscopic examination. Foreign bodies found in the anterior chamber following war wounds of traffic accidents were most commonly glass, quartz or stone. After removal of the foreign body, the endothelial dystrophy can heal spontaneously. A perforating keratoplasty of large diameter is indicated if the dystrophy persists or increases. The results are reported.


Spontaneous eosinophil-erythrocyte rosette formation. Eosinophils of dogs form rosettes with washed human erythrocytes in a manner very similar to that of thymus-derived (T) lymphocytes. Identification of rosette-forming eosinophils was made by both light and electron microscopic study of eosinophils infiltrating into the canine transmissible venereal sarcoma.


A case of fatal fat embolism syndrome following cardiopulmonary bypass. A case of fat embolism syndrome following median sternotomy and cardiopulmonary bypass, with fatal outcome, is described. This syndrome is present subclinically in the majority of these surgical procedures. The etiology and possible mechanisms of production are discussed; management is empirical and aimed at maintaining the function of embolized microvascular endothelium.


The assessment of function of left ventricle and patency of aorta-coronary bypass after operation. A study of systolic time intervals. The effect of aorta-coronary bypass surgery on left ventricular function was evaluated in 39 patients by cardiac catheterization and recording of the systolic time intervals (STI) before and an average of 4.5 months following myocardial revascularization. All patients were in sinus rhythm, had normal QRS intervals on the electrocardiogram, and had taken no medication for at least 3 weeks prior to the study. The STI was unchanged postoperatively in patients with the following characteristics: freedom from angina; heart rate not more than 15 beats greater than the preoperative level; all grafts patent; and no progression of disease in coronary arteries. Deterioration in left ventricular function after operation as represented by shortened left ventricular ejection time, (LVET) prolonged pre-ejection period (PEP), and increased ratio of PEP/LVET was found in patients with the following characteristics: persistent angina; heart rate more than 14 beats faster than the preoperative level; occlusion of any grafts; progression of disease in the coronary arteries; evidence for postoperative myocardial infarction. Worsening of left ventricular performance as determined by STI was more apparent in patients with occluded grafts and no collateral circulation than in those with occluded grafts and collateral circulation which may have offered protection against ischemia. These investigations supported the use of STI in assessing changes in left ventricular function after coronary bypass and, by inference, in assessing the status of the graft.


Tension on coronary bypass conduits. A neglected cause of real or potential obstruction of saphenous vein grafts. Of 41 saphenous vein aorta-coronary bypass grafts from 21 patients, abnormal graft tension was judged to be present at necropsy in 6 grafts (15 per cent) from 5 patients (24 per cent). Five (33 per cent) of 15 grafts placed to the right coronary system and one (3 per cent) of 26 grafts to the left coronary system were under excess tension. The abnormal tension appeared to result from the insertion of grafts too short to accommodate normal or abnormal distention of the right atrium and ventricle (graft to distal right coronary or posterior descending coronary artery) or of the pulmonary trunk (graft to left coronary system) after discontinuation of cardiopulmonary bypass or development of acute congestive cardiac failure.


Pulmonary involvement by histiocytosis X in the pediatric age group. The lungs may be involved by histiocytosis X either as part of a generalized disease or as a separate entity, primary pulmonary histiocytosis X. The former is much more frequent in the pediatric age group, and although it occurs primarily in those who are quite sick with widely disseminated disease, the lung involvement rarely caused symptoms and almost invariably clears. By contrast, primary pulmonary histiocytosis X is mainly seen in young adult males and frequently leads to severe disability. Twelve patients less than 20 years old with lung involvement by histiocytosis X are presented.


Human cochlear changes in noise induced hearing loss. The temporal bone histopathological findings in 14 ears with noise induced hearing loss are presented. The morphological changes consist mainly of hair cell loss, which is more severe in the 9 mm to 13 mm region of the cochlear duct. Within the area of maximum hair cell loss, there is a greater loss of outer hair cells than of inner hair cells. There is a good correlation between the permanent auditory threshold shifts and the spatial location of the sensory lesion according to the anatomical frequency scale.


Analgesic nephropathy.

Analgesic nephropathy. Analgesic nephropathy is part of a wider clinical syndrome associated with the abuse of APC compounds, that is, a minimum total intake of 2 kg of aspirin or phenacetin. Ischaemic heart disease and premature aging are newly recognized aspects of the analgesic syndrome. The diagnosis of analgesic nephropathy can be made precisely by the radiological demonstration of renal papillary necrosis. The most important aspect of management of established analgesic nephropathy and renal insufficency is total avoidance of all non-steroid antiinflammatory agents and this is commonly associated with stabilization or improvement in renal function. In the APC mixture, aspirin appears to be the major nephrotoxic agent while phenacetin and paracetamol play a secondary and synergistic role in the nephrotoxicity.


Methotrexate megadosage followed by folate rescue. I. A study in clearance patterns. Plasma clearance rates of methotrexate  (MTX) have been determined in five patients given an infusion of 3,000 mg of MTX over six hours, followed by folate rescue 24 hours after the commencement of infusion. Results presented indicate that maintenance of high MTX concentrations and potential toxicity could not be related to age, body weight or surface area.


Localized tissue reduction.

Localized tissue reduction. A faradic muscle  stimulator (the Hawkins EBA machine) for localized tissue reduction is described. Forty patients undergoing a course of treatment with the machine all lost significant amounts of weight. No side effects were noted.


Methotrexate megadose followed by folate rescue. II. Clearance patterns in patients receiving sequential megadose infusions. In some patients receiving megadose infusions of methotrexate (MTX), the rate of drug clearance decreases with each subsequent infusion. This can be modified by administration of fluids as outlined in the Recommended Protocol. Monitoring of each treatment is needed to assess requirements of fluids since these cannot be adequately predicted from data of previous infusions.


[Extracorporeal blood irradiation in chronic lymphatic leukemia (author's transl)]. In 8 outpatients suffering from different stages of chronic lymphatic leukemia extracorporeal irradiation of blood was used. In two cases full remission could be induced. In the other patients leukocytes were reduced to 10000 to 40000. In all patients we found a diminuation of B lymphocytes. Side effects were not observed.


Receptors for sheep erythrocytes (E) and erythrocyte-antibody-complement complexes (EAC) on the lymphoblasts of childhood acute lymphocyblastic leukemia. Two cell-surface markers, rosette formation with sheep erythrocytes (E-rosette) as a T-cell marker and rosette formation with bovine erythrocyte-antibody-complement complex (EAC-rosette) as a B-cell marker were determined on peripheral blood lymphocytes and lymphoblasts from normal and 89 children with acute lymphoblastic leukemia (ALL). In the majority of the patients (12/15 untreated patients and 6/11 patients in relapse), lymphoblasts exhibited neither E- nor EAC-rosette formation. Lymphoblasts from one untreated patient with mediastinal mass displayed E-(50%) and EAC-rosette formation (15%). In 3 of 11 patients in relapse, lymphoblasts displayed an increase in EAC-rosette formation with progressive disease. In the remaining patients with active disease, a small and variable proportion of lymphoblasts expressed E and/or EAC-rosette formation. In 63 patients in remission, percentages of E- and/or EAC-rosette were similar (p greater than 0.05) to those of control. The results indicate a wide heterogeniety with respect to expression of lymphocyte membrane markers in lymphoblasts and in normal lymphocytes in patients with active ALL.


Bacteriophage N3 of Haemophilus influenzae. I. Independence of vegetative recombination among Haemophilus influenzae bacteriophage on the host cell. Vegtative recombination of temperature-sensitive mutants of Haemophilus influezae bacteriophage N3 was measured in wild-type H. influenzae strain Rd9 and in recombination-defective mutants of the Rd strain. Recombinants are formed with low but equal frequency in wild-type cells and recombination-defective mutants of the Rd strain. It is concluded that this phage can carry information in its own genome for vegetative recombination. Lysogenization readily takes place in both strains.


Phospholipids bound to acidic nuclear proteins in human B and T lymphocytes. Chromatin fractions (DNA, histones and non-histone chromosomal proteins NHCP) have been isolated from human peripheral B and T lymphocytes using different methods and analyzed in order to identify their lipid content. While DNA and histone fractions do not reveal the presence of lipids, a 2% of phospholipids is present in the NHCP fraction. The phospholipids associated with NHCP present a constant relative ratio among sphingomyelin, phosphatidyl-choline and phosphatidyl-ethanolamine both in B and T lymphocytes, whichever are the extraction procedures employed. These findings are related to the possible dereprssive role of phospholipids on DNA-dependent RNA synthesis.


The effect of electrical stimulation and drugs on the release of acetylcholine from the frog spinal cord. The isolated frog spinal cord preparation, hemisected longitudinally, has been used to study concurrently the release of endogenous acetylcholine (ACh) and the electrical responses to stimulation of the ventral or dorsal roots. In unstimulated cords, ACh release was high initially and then fell to a plateau level. ACh release was reduced by tetrodotoxin and increased by high K+. Stimulation of the ventral roots enhanced the ACh release. The magnitude of this effect was dependent on the frequency of stimulation and was antagonized by tetrodotoxin or C Ca2+-free Ringer. During prolonged ventral root stimulation (50 min), the evoked ACh release became progressively smaller but was restored to the original high levels by the addition of choline to the bath fluid. Stimulation of the dorsal roots failed to augment the ACh output. The application of hyoscine was followed by a large rise in the unstimulated ACh output. However no increase in the antidromically-evoked release, when compared with control stimulations, was seen. The application on alpha-bungarotoxin was followed by a short-lasting decline in the unstimulated ACh output, whereas the electrically-evoked output was unaffected. In the frog spinal cord the determination of ACh release, together with the recording of the root potentials, offers the possibility of studying the output of a central transmitter in relation to the electrical activity of an in vitro central nervous system preparation.


[A clinicopathological study of the cranial "histiocytosis X" (AUTHOR'S TRANSL)]. A clinicopathological study of 13 cases of cranial histiocytosis X was discussed. The patients were mostly infants, and those under 15 years of age numbered eight. Hematological, blood chemistrical, and other laboratorical prodedures have produced no direct diagnostic value. Radiographically, this lesion in skull appeared to be a lytic defect in all cases. Histologically, they were unified by a localized or generalized histiocytic proliferation. Eosinophilic cells were also found throughout the lesional tissue. There were some slight histological variations from case to case, for instance, in the degree of infiltration by eosinophilic cells, and the number of the histiocytic cells. However, the basic histological pattern remained the same in all cases. The lesion may already have extended into the neighboring osseous tissues. Sections of the neighboring osseous tissues showed the intertrabecular spaces to be infiltrated by the same type of lesion. The patient related trauma to the onset of symptoms in 6 cases, but it is true that we know nothing of the etiology of this lesion. The treatment of choice is the adequate surgical extirpation. It seems fair to conclude that wide surgical excision with postoperative irradiation or with chemotherapy may be expected to affect a clinical cure.


The medial posterior choroidal artery as an indicator of masses at the foramen of Monro. The angiographic findings in five cases of tumors at the foramen of Monro are presented. Characteristic changes in the course and configuration of the medial posterior choroidal artery are described.


B and T lymphocytes in man. III. B, t, and "null" lymphocytes in multiple sclerosis. T, B, and "null" lymphocytes were determined in peripheral blood obtained from 24 control subjects and 15 multiple sclerosis patients. The total number of lymphocytes and T lymphocytes were reduced significantly in multiple sclerosis patients as compared with those of controls. Although the percentage of B lymphocytes increased in these patients, the absolute number of B lymphocytes was similar to controls. The mitotic response of lymphocytes from multiple sclerosis patients to phytohemagglutinin and pokeweed mitogen in 7-day-old cultures was decreased as compared with that of controls.


Malignant lymphoma and rheumatic symptoms. A patient intially thought to have a connective tissue disease, later shown by biopsy to have Hodgkin's disease is described. The progress of her illness suggests that she is suffering from a connective tissue disease, too, or possibly secondary immune complex disease.


Mandibular histiocytosis X and acute lymphoblastic leukemia. The occurrence together of two major distinct diseases in the oral area is relatively rare. In this report, a case is presented in which histiocytosis X was found to develop in a child with acute lymphoblastic leukemia. The nature of the differentiated histiocytoses is explored, and the possibility of the relationship between a lymphoproliferative and a histiocytic disease entity is investigated.


Evaluation of a rapid beta-lactamase test for detecting ampicillin-resistant strains of Hemophilus influenzae type b. Chloramphenicol is presently the drug of choice in the initial treatment of serious infections due to Hemophilus influenzae type b. Rapid detection of ampicillin resistance in clinical isolates would facilitate early discontinuation of chloramphenicol therapy in patients infected with ampicillin-sensitive bacteria. A total of 160 strains of H. influenzae type b were tested with a one-hour acidimetric microassay for beta-lactamase activity. All ampicillin-resistant strains rapidly hydrolysed the beta-lactam ring of penicillin. When isolates were encoded and tested without knowledge of their MICs, the 40 ampicillin-resistant strains (MIC greater than or equal to 2 mug/ml) were readily distinguished from 120 sensitive strains. Rapid beta-lactamase assay is therefore a reliable detector of ampicillin resistance in H. influenzae type b.


Epidemiologic characteristics of infections caused by ampicillin-resistant Hemophilus influenzae. Hrom February 1974 through May 1975, 58 isolates of ampicillin-resistant Hemophilus influenzae from patients were submitted to the Center for Disease Control from 23 states and the District of Columbia. The epidemiologic characteristics of these widely scattered cases were strikingly similar to those of cases previously reported from individual communities or institutions. Because of the nationwide distribution of resistant isolates and the adverse consequences of ineffective therapy, the use of chloramphenicol is recommended for the initial management of systemic illness definitely or probably due to H. influenzae type b.


Asymmetrical displacement current and its relation with the activation of sodium current in the membrane of frog myelinated nerve. 1. Sodium currents (INa) and asymmetrical displacement currents (ID) were measured in the same nerve fibres from Rana esculenta under similar conditions. 2. For exploring possible kinetic and steady state relations between INa and ID the following quantities were compared: (i) the activation of the sodium channels and (ii) the charge displacement of ID. 3. The delay of sodium activation increased after hyperpolarization. A corresponding effect on the displacement of charge was not observed. 4. Upon a small depolarization sodium activation rose slower than the displacement of charge, whereas at large depolarizations sodium activation reached a steady level before the charge displacement. 5. Upon repolarization to various levels between -52 and 12 mV relative to the resting potential, the ratio between the time constants of charge displacement and of sodium tail current varied between 3 and 1. 6. In the steady state the sodium activation was one half at about the same potential as the charge displacement but exhibited a clearly steeper voltage dependence. 7. Blocage of sodium channels with tetrodotoxin did not affect the asymmetrical displacement current. Replacing a part of external Na by tris did not alter the sodijm activation process. 8. The results indicate that the asymmetrical displacement of charge may reflect states of the gating mechanism in sodium channels but cannot be considered as a correlate of the Hodgkin Huxley m variable.


Saxitoxin and procaine act independently on separate sites of the sodium channel. 1. Voltage clamp experiments were done on single myelinated nerve fibres of the frog, Rana esculenta. 2. The time course of procaine action (1.0 mM at pH 7.2) was obtained from changes in INa on changing solutions during repetitive (1 HZ) depolarizing pulses of constant amplitude following hyperpolarizing prepulses. The mean half times of onset and offset of procaine block were 3.7 and 28 s, respectively. In the presence of 1.4 nM saxitoxin (STX) the corresponding times were virtually the same, 3.1 and 27 s. 3. Similarly, the time course of partial relief from procaine block that is obtained by increasing the frequency of the prepulse-test pulse pairs from 1-10 HZ was unaffected in the presence of STX. 4. Comparison of the equilibrium effects of procaine concentrations ranging from 0.03-1.0 mM suggest a one-to-one drug-receptor reaction. The fraction of Na channels blocked at equilibrium with 1.0 mM procaine, 1.4 nM STX, and 1.0 mM procaine + 1.4 nM STX was 0.81, 0.49, and 0.90, respectively. This result and the kinetic behaviour fully agree with the idea of two separate and independent receptors for procaine and STX.


Studies on the relation between latency relaxation and resting cross-bridges of frog skeletal muscle. Latency relaxation of frog skeletal muscle (LR) was investigated with respect to its relation to resting cross-bridges. A decrease in the initial stiffness of the resting muscle (stiffness of the muscle during the beginning of a length-change) was found, when repeated triangular length-changes were imposed on the muscle. This decrease in the initial stiffness depends on the velocity of the length-change. It is interpreted that the decrease in the initial stiffness reflects a detachment of the resting cross-bridges from their binding-sites. The LR, induced immediately after the offset of the length-changes, i.e. when the cross-bridges are still detached, showed an increased depth, its time course remaining unchanged. There is a strong correlation between the increase in the depth of the LR and the decrease in the initial stiffness. The LR regained its original depth (depth without a preceding length-change) about 5 s after the offset of the length-changes (20 degrees C). It is suggested that the LR of skeletal muscle is not due to a detachment of resting cross-bridges.


Renal test dyes III. Effect of dyes suitable for renal passage time measurements on renal function. The effects of the triaryl-methane dyes Lissamine Green SF (LF SF). Lissamine Green BN (LG BN), Lissamine Green V (LG V) and Kiton Pure Blue V (KPB V) on renal functions of rats and on sodium transport across isolated frog skin have been investigated. The experiments failed to show any natriuretic or diuretic effects of the purified dyes on the rat kidney. In the continuous infusion experiments, which lasted 3 1/2 h, the amount of the dye infused was equivalent to a passage time measurement every 3 min. In rapid injection experiments LG V was injected every 5 min for 1 h. The dyes did not have any effect on insulin clearance either. Previously reported effects of the dyes can be partly explained by the presence of sodium sulfate in commercially available dyes. Experiments with LG V showed that the dye also has no effect on the potential difference or short circuit current across the isolated frog skin. This supports their usefulness as renal test dyes.


Aplastic anemia: presence in human bone marrow of cells that suppress myelopoiesis. Bone marrow from a patient with aplastic anemia was shown by multiple criteria to have a block in early myeloid differentiation. This block was overcome in vitro by elimination of marrow lymphocytes. Furthermore, this differentiation block was transferred in vitro to normal marrow by coculturing with the patient's marrow. We suggest that some cases of aplastic anemia may be due to an immunologically based suppression of marrow cell differentiation rather than to a defect in stem cells or their necessary inductive environment.


Von Willebrand's disease--a potential bleeding problem in plastic surgery. Von Willebrand's disease, a common hereditary bleeding disorder, may be responsible for undue or unanticipated intraoperative or postoperative hemorrhage. The diagnosis may be difficult to establish, although it may be suspected clinically and can usually be confirmed by adequate laboratory investigation. Patients with Von Willebrand's disease appear to be especially susceptible to developing significant prolongation of their bleeding time after the ingestion of aspirin.


Effects of tandamine and pirandamine, new potential antidepressants, on the brain uptake of norepinephrine and 5-hydroxytryptamine and related activities. Two novel agents, tandamine (TA; a thiopyrano (3,4-b) indole) and pirandamine (PA; an indeno (2,1-c)pyran), and the tricyclic antidepressants desimipramine (DMI), imipramine (I) and amitriptyline (A) were compared in various in vivo pharmacological tests and for norepinephrine (NE) and 5-hydroxytryptamine (5-HT) neuronal uptake inhibition. TA was found to be equivalent, or greater, in activity to DMI in blocking brain NE uptake, antagonizing reserpine-induced effects and potentiating the behavioural effects of l-Dopa. Similarly to DMI, TA did not appreciably block brain 5-HT uptake; unlike DMI, TA did potentiate central 5-HT activity at high doses. PA exerted an opposite profile to TA, being equivalent to A and greater than I as a 5-HT uptake blocker and central 5-HT potentiator; PA was not effective as a NE uptake blocker or potentiator. Neither TA or PA exhibited in vivo MAO inhibition, and in contrast to DMI, I and A, exhibited no central anticholinergic effects. TA, but not PA, potentiated apomorphine-induced gnawing. These findings indicate that TA is a relatively specific blocker of neuronal NE uptake and PA is a selective 5-HT uptake blocker.


The usefulness of computed tomography during and after radiation therapy in patients with brain tumors. Two years of experience with 100 patients in the serial study of brain tumors during and after radiation therapy has shown that computed tomography is useful in (a) depiction of regression or growth of primary and metastatic intracranial tumors, (b) recognition of untoward sequelae of radiation therapy, such as necrosis and edema, and (c) appreciation of changes in tumor density and ventricular size. Representation cases are discussed and illustrated.


Computed tomography in the evaluation of malignant glioma before and after therapy. Sixty-three patients with malignant glioma undergoing chemotherapy and radiotherapy were evaluated with electroencephalography, 99mTc-DTPA imaging, and computed tomography (CT). Tumor size, central lucency, contrast enhancement, surrounding edema, and ventricular size were assessed. CT findings were found to be reliable and often predicted the clinical course. Tumor size, central lucency, and contrast enhancement increased in patients with clinical deterioration and decreased in those with improvement. The CT scan also provided additional information of value in adjunctive therapy, such as the presence of cysts or ventricular obstruction. In general, CT gave a more complete profile of tumor characteristics than other diagnostic modalities.


Air pollution in copenhagen. Part I. Element analysis and size distribution of aerosols. Aerosols were monitored in Greater Copenhagen in the period June 1973 to July 1974. Size-fractionated cascade impactor samples and unfractionated filter samples were regularly collected and analyzed by neutron activation analysis, spark emission spectroscopy or proton-induced X-ray emission spectroscopy. Concentrations were determined of the following elements: Al, Si, S, Cl, K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, Br, Mo, Cd, Sn, Sb, and Pb. All elements showed orders-of-magnitude fluctuations; the mean concentrations were roughly the same as in other large cities. In relation to proposed air quality standards, Pb was the most critical component. Statistical analysis of variation patterns, size distributions and interelement correlations indicate that automotive exhaust is the source of Br and Pb; fuel-oil combustion is the main source of V and Ni (and partly of S); soil dust raised by wind or by human activity (e. g. traffic) is the main source of Al, Si, Ca, Ti, and Fe.


[Angiographic diagnosis of localized gastro-Intestinal hemorrhage (author's transl)]. Origin and site of gastro-intestinal hemorrhage in 3 cases of acute and 14 cases of chronic bleeding are described. In such cases of angiodysplasia other investigations and, in some, even repeated operation could not find the origin, but angiography succeeded.


Modification of the response of human T lymphocytes to phytomitogens by cocultivation with unresponsive non-T leukocytes. Experiments were conducted to ascertain to what extent the mitogen response of human peripheral T cells is influenced by contaminated, nonresponding leukocytes. T- and B-lymphocyte preparations, extensively depleted of monocytes, exhibited extremely poor responses to pokeweed mitogen and concanavalin A compared with the original unfractionated cell population. A strongly reduced phytohemagglutinin response was noted in the B-cell but not in the T-cell fraction. Mixtures of cell preparations enriched in T or B cells yielded stimulations higher than expected. Mitomycin-C-treated T-cell preparations mixed with untreated preparations enriched in B cells gave slightly higher responses than expected. However, highly enhanced responses were observed when untreated T cells were mixed with mitomycin-treated cell suspensions enriched in B cells. A similar enhancing effect was noted when mitomycin-treated cell suspensions enriched in adherent cells were added. It is concluded that the mitogen reactivity of T cells, on a cell-for-cell basis, may be enhanced tenfold or more by cocultivation with unresponsive non-T leukocytes. A reduced mitogen reactivity of peripheral lymphoid cells from patients with various types of disease may thus either reflect a decreased proportion of responsive lymphocytes or unresponsive non-T lymphocytes capable of enhancing mitogen reactivity, or both.


[Partial syndrome of cerebello pontine region due to an aneurysm of the internal auditory artery (author's transl)]. An aneurysm of the distal portion of the middle cerebellar artery was first clipped and then, given the type of neck, removed. The case is reported because of its extreme rarity, the unusual succession of symptoms, the difficulty of diagnosing the malformation, (thanks to modern radiological techniques) and the successful outcome of surgery.


Binding of IgM by human T lymphocytes.

Binding of IgM by human T lymphocytes. Ox erythrocytes, sensitized with IgM-type rabbit antibodies, were found to form rosettes with human blood lymphocytes. Granulocytes, monocytes, and macrophages appeared to lack this property. The rosette formation could be inhibited with human IgM and its Fc fragments. IgM-binding lymphocytes were also present in cell suspensions isolated from human tonsils and thymuses and among blood lymphocytes cultured in vitro. The outcome of combined marker tests and cell separation experiments disclosed that the rosetting lymphocytes belong to the T-lymphocyte population.


Characterization of dextran-sulfate-sensitive cells. The cell type participating in the mitotic response to the polyclonal B-cell activator (PBA) dextran-sulfate (DS) was investigated. Cells from murine fetal liver, adult bone marrow, and spleen were studied; only a limited number of all cells present in each organ responded to DS. Morphological studies of the activated cells showed the major population of activated cells in spleen to have the appearance of lymphoblasts. In bone marrow, several classes of hematopoietic cells were mitotically active, including mononuclear cells (lymphoblasts and monocytes), megakaryocytes, and myeloblasts. In these cultures, however, it was not possible to differentiate between DS-activated and spontaneously proliferating cells. Bone marrow and, to some extent, spleen cell cultures activated with DS contained a relative increase in numbers of phagocytic cells, whereas stimulation of spleen cells with lipopolysaccharide did not result in an increased phagocytosis. However, adherent cells were not necessary for activation of DNA synthesis by DS in spleen, and this cell type did not contribute to a measurable degree to the DNA synthetical response. DS cannot be regarded as a general stem cell mitogen for bone marrow cells since it failed to promote colony growth of hematopoietic cells in an in vitro system. However, supernatants from DS-activated spleen and bone marrow cell cultures did stimulate colony growth of murine bone marrow cells, indicating that stem cells of nonlymphoid origin might be indirectly activated by DS. In conclusion, the major cell population activated by DS in spleen is lymphocytes. In bone marrow, other cell types seem to participate in the response as well, but the activation mechanism may be indirect and not primarily the result of DS interaction with these cells.


Off-signals in lymphocyte activation.

Off-signals in lymphocyte activation. High-dose unresponsiveness to concanavalin A affected T and B lymphocytes equally, although only T cells were activated by Con A. Thus, activation and inactivation signals are totally unrelated, the latter also lacking target cell specificity. The off-signal caused cell death and is probably biologically unimportant.