Fluorescent staining of fungal nuclei with a benzimidazol derivative. A direct staining procedure is described for fluorescence microscopy of fungal nuclei, chromosomes and mitochondria. The fluorochrome is a benzimidazol derivative (33258 Hoechst) known to bind selectively to deoxyribonucleic acid at neutral pH. The advantages of 33258 Hoechst  relative to Feulgen compounds used previously to stain these structures include a greater intensity of fluorescence, the absence of fading or rapid quenching of the fluorescence, and the omission of acid hydrolysis from the procedure for removal of ribonucleic acid. 33258 Hoechst has been evaluated as a nuclear stain with a yeast (Saccharomyces cerevisiae) and a filamentous fungus (Agaricus bisporus) and appears to penetrate easily vegetative cells and spores of both fungi.


Development and evaluation of an immunodiffusion test for diagnosis of systemic zygomycosis (mucormycosis): preliminary report. An antigen analysis with filtrate and homogenate precipitinogens of single isolates of the zygomycetes Absidia corymbifera, Mucor pusillus, Rhizopus arrhizus, and Rhizopus oryzae demonstrated the presence of common antigens among the three genera as well as antigens which permit their differentiation. Selected homogenate antigens were valuable in developing a diagnostic immunodiffusion (ID) test for systemic zygomycosis. When sera from 43 patients with various proven mycoses other than zygomycosis were tested against each of the antigens, none formed precipitin bands identical to those formed by A. cormybifera, M. pusillus, and the Rhizopus spp. rabbit reference antisera. Sera from 23 normal persons and 25 diabetics did not react with any of the antigens. Homogenate antigens detected antibody in 8 of the 11 sera (73%) from suspected or proven cases of zygomycosis, whereas ID tests with filtrate antigens detected antibody in only 2 of the 11 sera (18%). Of the eight sera that reacted with the homogenate antigens, five only reacted with a specific Rhizopus sp. antigen, two only reacted with a specific M. pusillus antigen, and one only reacted with a specific A. corymbifera antigen. Study results show the ID test with homogenate antigens to be more specific and sensitive than the ID test with filtrate antigens and indicate that the former is a promising technique for diagnosing human zygomycosis.


Alpha1-fetoprotein in the diagnosis of hepatoma: statistical and cost benefit aspects. A rational comparison of different serum concentrations of alpha1-fetoprotein (S-AFP) in the diagnosis of hepatoma must be made. We took data on the sensitivity and specificity of different diagnostic S-AFP concentrations from the literature and evaluated them statistically and by Bayesian analysis. In our patients (hepatoma prevalence 0.028) a sensitive diagnostic concentration (30-50 ng/ml) will misdiagnose hepatoma so often that a positive test will indicate hepatoma in only 10% of cases. A positive test at a specific diagnostic concentration (500 ng/ml) indicates hepatoma in 100% of cases and is preferable in terms of cost benefit. Although the lower concentration will diagnose a larger proportion of patients with hepatoma (74% compared with 59%) the 'costs' of excluding false positives are considerable (A$2545 per extra case with 2.5% of patients suffering significant morbidity). In western societies, where the prevalence of hepatoma is low, a higher, less sensitive but more specific diagnostic S-AFP concentration is appropriate.


Cross contamination of cytological smears, with automated staining machines and bulk manual staining procedures. With a specific study of the problems of the Cytotek and the Shandon Elliott staining machines. Further development of an individual staining machine is to be strongly encouraged but meanwhile, using bulk stainers, frequent changing of wash fluids and staining solutions, particularly leading up to and following the haematoxylin pot, is essential to reduce the risk of cross contamination. Certain smears, such as from semen or from serous fluids where malignancy is suspected or known, must be stained on separate racks. In some laboratories it is the rule not to stain semen or serous fluids in bulk staining machines at all and this may have to become the rule everywhere until we are provided with safe individual slide stainers.


Developmental retardation in infants as a concomitant of physical child abuse. The impact of child abuse on the developmental functioning of infants was investigated. Thirty verified cases of physically abused children were compared to a reference group of 30 nonabused children matched for age, sex, race, and socioeconomic status. Abused children scored significantly lower in terms of cognitive and motor development as measured by the Bayley Scales of Infant Development. Development delays on three of the four sectors of the Denver Developmental Screening Test, personal-social, language, and gross motor, were also found in the abused children. There were, however, relatively few item differences between the two groups on the 30 more general behavioral variables constituting the Bayley Infant Behavior Record. Results appear to confirm clinical observation of abused children as developmentally retarded with specific delays in the language and gross motor areas. Although methodologically complex, longitudinal studies are clearly indicated to assess the stability and/or reversibility of the present findings.


Tissue fixation and staining with osmium tetroxide: the role of phenolic compounds. It has been postulated that phenol-containing areas of plant and animal tissues were osmiophilic, but proof of direct interaction between osmium tetroxide and phenolic materials, or the nature of such reactions, has been lacking. We find that, under conditions similar to those of normal tissue fixation, osmium tetroxide reacts rapidly with those phenols containing o-dihydroxy groups (including such species found in plant tissues) to give very stable chelate complexes. We conclude that these complexes are responsible for the observed electron-density in phenol-containing areas of tissue treated with osmium tetroxide, so that such phenols are indeed osmiophilic.


Investigation of the antigenic composition of the cells and the exotoxin of Sh. dysenteriae (Shiga) by agar immunoelectrophoresis and polyacrylamide gel disc electrophoresis. Using the method of immunoelectrophoresis in agar, we established the presence of 12 soluble antigens in Sh. dysenteriae (Shiga). We differentiated among them 3 specific antigens situated in the cathodic region. One of them is the somatic O antigen while the other two are thermolabile surface K antigens of these bacteria. K antigens of Grigoryev-Shiga bacteria differ from the earlier described K antigens of the Newcastle and Boyd Shigellae in their positive electric charge, in greater lability in relation to temperature and chemical effects and evidently also in their chemical character. The remaining protein antigens are common for either the genus of Shigella or for the family of enterobacteria. The exotoxin preparation isolated from the R strain of Sh. dysenteriae (Voile 30) by precipitation with trichloroacetic acid is represented in the immunophoreogram by five precipitation lines. Disc electrophoresis in polyacrylamide gel made possible the investigation of the composition of the exotoxin and isolation of the two fractions responsible for its toxic effect. The method of preparative electrophoresis in polyacrylamide gel can evidently be used for obtaining purified Sh. dysenteriae exotoxin preparations.


Tolerance to azobenzenearsonate: preferential loss of the major normal cross-reactive idiotype. A specific tolerant state was induced in A/J mice to the hapten p-aminobenzenearsonate (Ars) by the injection of deaggregated conjugates of Ars and human gemma-globulin (Ars-DHGG). The kinetics of tolerance to Ars-DHGG was found to be identical to that of B cell tolerance to human gamma-globulin alone. The ability to produce antibody (anti-Ars) bearing the major normal cross-reactive idiotype was found to be preferentially lost. In addition, recovery of the ability to produce anti-Ars bearing this cross-reactive idiotype was found to be delayed as compared to the total anti-Ars response upon spontaneous loss of tolerance.


Specific suppression of the immune response by a factor obtained from spleen cells of mice tolerant to human gamma-globulin. An antigen-specific suppressive factor was extracted from spleen cells of mice made tolerant by injection of deaggregated human gamma-globulin (HGG). The active material could be prepared from T cells, obtained by passaging spleen cells through an anti-immunoglobulin column, although not from cells adherent to the column nor from spleen cells pretreated with anti-Thy-1 serum and C. This factor was antigen-specific since it was retained on immunoadsorbents containing HGG, but not on columns coated with antibody to HGG or with irrelevant antigens. Despite its specificity for antigen it did not bear any classical immunoglobulin determinants. Its m.w. ranged between 30,000 and 55,000 daltons. It was a product of the I region of the major histocompatibility complex since it carried Ia determinants. The properties of this factor are very similar to those reported elsewhere for suppressive factors obtained from primed T cells, cells from nonresponder mice, and allotype-specific cells. This suggest the existence of a major class of immunoregulatory molecules, nonimmunoglobulin in nature, and responsible for the mediation of antigen-specific T cell-dependent suppression.


Immune response to phosphorylcholine. I.V. Comparison of homologous and isologous anti-idiotypic antibody. Homologous and isologous anti-idiotypic antibodies against the PC-binding BALB/c myeloma protein HOPC-8  were raised in A/He and BALB/c mice, respectively. Isologous and homologous anti-HOPC-8 serum suppressed the response to PC in vitro specifically. The antibodies were purified by using HOPC-8 immunoabsorbent. Purified isologous and homologous anti-idiotypic antibody were labeled with 125I and compared for Ig class composition and idiotype-binding specificity. Both kinds of anti-idiotypic antibodies were predominantly IgG1, were highly specific for the HOPC-8 idiotype, and had a smiliar affinities for the PC-binding site. These findings demonstrate that anti-HOPC-8 antibodies raised in A/He and BALB/c mice are very similar in their biologic and immunochemical poperties.


Immunologic responses to a murine mammary adenocarcinoma: in vitro production of specific killer cells is dependent on active T lymphocytes. The mode of production of specifically armed monocytic killer cells was investigated with the T1699 mammary adenocarcinoma in syngeneic DBA/2 mice. After overnight in vitro incubation of cells from the spleen but not from the lymph nodes, blood, or from the peritoneal cavity produced specific killer cells. The activation of spleen cells was inhibited by pretreatment with anti-theta serum and C; however, already activated specific killer cells were not sensitive to the same treatment. Removal of phagocytic cells did not significantly affect the cytotoxicity of the splenic killer cells whereas removal of rayon-wool adherent cells greatly reduced both the total cytotoxicity, and to a lesser extent, the cytotoxicity indices. Overnight co-cultivation of normal peritoneal-exudate cells with the lymph node cells from tumor-bearers, although neither class of cells alone was cytotoxic to T1699 cells in vitro, produced specific monocytic killer cells, through steps dependent on active T lymphocyte function. Culture spupernatants of tumor-bearer's spleen cells also contained factor(s) which induced cytotoxicity mediated by normal peritoneal-exudate cells against T1699 cells in vitro; and the production of the factor(s) was also inhibited by pretreatment of the spleen cells with anti-theta serum but not by anti-mouse IgG or anti-mouse whole gamma-globulins serum and C.


On the cross-reactivity of staphylococcal enterotoxins A, B, and C. Strong cross-reactions were demonstrated for staphylococcal enterotoxins B (SEB) and C1 (SEC1) by antigen-binding capacity and by competitive binding ability. Both SEB and SEC1 combined completely with the heterologous antibody although requiring four times as much antiserum as the homologous enterotoxin and both displaced about one-third of the other enterotoxin from a heterologous antigen-antibody system. It is proposed that one of the three major antigenic determinants of these enterotoxins possesses a significant similarity but probably not an identity of structure. SEB and SEC1 did not combine with antiserum to enterotoxin A nor inhibit the reaction of SEA with anti-SEA. SEA had no intrinsic binding capacity for anti-SEB or anti SEC1 nor did it inhibit the binding of either enterotoxin to its own antibody. Affinity chromatography was employed to demonstrate that a small apparent binding of SEA to anti-SEB was due to antibody to SEA in the anti-SEB serum and that an almost complete displacement of SEC1 binding to anti-SEC1 was caused by contaminating SEC (about 0.01%) in preparations of enterotoxin A.


Differential expression of an equivalent clonotype among BALB/c and C57BL/6 mice. The primary anti-phosphorylcholine (PC) response in BALB/c, C57BL/6, and congenic and recombinant inbred strains of these parental types has been examined in the splenic focus system. The frequencies of PC-specific precursors were shown to vary among these strains from 2 to 20 precursors per 10(6) splenic B cells. The distribution of these frequencies suggests that elements closely linked to or within the major histocompatibility complex may play a role in the determination of this parameter, although additional experiments are necessary to adequately assess this possibility. Moreover, all strains tested, regardless of immunoglobulin allotype, expressed monoclonal antibodies indistinguishable from the TEPC 15 myeloma protein (T15) clonotype. Further, the frequency of this clonotype in a given strain did not appear related to allotype, since both high and low T15 frequencies were found among strains of either the BALB/c (a(1)) or C57BL/6 (a(2)) allotype. The examination of normal serum for the T15 idiotype, however, revealed that only mice of the BALB/c allotype (a(1)) expressed the T15 idiotype in detectable quantities. After immunization with Diplococcus pneumoniae, sera from mice of the a(1) allotype consistently contained large quantities of the T15 idiotype, whereas sera from mice of the a(2) allotype exhibited various degrees of cross-reactivity with anti-T15 antibody. These results suggest that: (a) the allotype of an individual, although closely related to serum levels of an idiotype, is unrelated to the proportion of the precursor population which expresses that idiotype and; (b) the serum expression of a given idiotype may reflect regulatory processes, which act either during or before antigenic stimulation, rather than the actual clonotype representation in the repertoire. These findings indicate that distinctions must be made between the expression of idiotypic determinants within precursor B-cell populations and elements which regulate the subsequent appearance of those idiotypes in serum antibodies.


Antigen-specific T-cell-mediated suppression. I. Induction of L-glutamic acid60-L-alanine30-L-tyrosine10 specific suppressor T cells in vitro requires both antigen-specific T-cell-suppressor factor and antigen. A combination of in vitro and in vivo techniques were used to explore the mode of action of both crude and purified suppressive extracts specific for the random copolymer L-giutamic acid(60)-L-alanine(30)-L-tyrosine(10) (GAT- T(s)F) obtained from nonresponder DBA/1 (H-2(q)) mice. Normal DBA/1 spleen cells were incubated under modified Mishell-Dutton culture conditions for 2 days together with crude or purified GAT-T(s)F, and in the presence or absence of free GAT. These cells were then washed extensively and 3 x 10(6) viable cells transferred to syngeneic recipients, which were challenged at the same time with the immunogenic form of GAT complexed to methylated bovine serum albumin (GAT-MBSA). GAT-specific IgG plaque-forming cells (PFC) in the spleen were assayed 7 days later. In agreement with earlier in vitro studies on the action of GAT-T(s)F, it was demonstrated that under these conditions, low concentrations of GAT-T(s)F stimulated the development of cells which, aider transfer, are able to suppress the GAT PFC response to GAT-MBSA. The cells responsible for this suppression were shown to be T lymphocytes by using nylon wool-purified T cells for suppressor cell induction and by eliminating suppressive activity in cells cultured with crude GAT-T(s)F by treatment with anti-Thy 1.2 plus C before transfer. The suppressor T cells act in a specific manner failing to suppress significantly either anti-sheep erythrocyte or trinitrophenyl-ovalbumin primary PFC responses. For the induction of GAT-specific suppressor T cells in culture, a moiety bearing H- 2(K(q) or I(q)) determinants and also GAT, either bound to the crude GAT- T(s)F or added in nanogram amounts to antigen (GAT)-free purified GAT-T(s)F, were both required.


A suppressor T cell of the mixed lymphocyte reaction in man specific for the stimulating alloantigen. Evidence that identity at HLA-D between suppressor and responder is required for suppression. It has previously been shown that J.H., a human leukocyte antigen (HLA)-Dw2 homozygous multiparous woman, fails to respond in a mixed lymphocyte reaction (MLR) to her Dw1 homozygous husband W.H., and that her T cells suppress the responses of HLA matched responders to W.H. The present studies take advantage of the observation that J.H. suppressor cells resist a dose of gamma-irradiation which functionally eliminates her MLR responder cells. J.H. cells, depleted of alloreactive cells, suppress the responses of Dw2 heterozygous or homozygous cells to W.H., regardless of their associated HLA-A or B antigens. Only when W.H. or a few other cells are present as the irradiated stimulator is J.H. suppression of Dw2 responses detected. Thus, the J.H. suppressor T cell recognizes determinants in the irradiated stimulator cells as well as D locus products in the responder.


Ontogeny of B-lymphocyte function. V. Thymus cell involvement in the functional maturation of B-lymphocytes from fetal mice transferred into adult irradiated hosts. Lethally irradiated mice reconstituted with adult thymus cells and neonatal or fetal liver cells produce an anti-2,4-dinitrophenyl or anti-bovine gamma globulin response of restricted heterogeneity of affinity in comparison with the response of mice reconstituted with B cells from adult donors. In addition, mice reconstituted with day 15 fetal B cells and adult thymus cells produce relatively few indirect plaque-forming cells (PFC). It was found that B cells acquire the capacity to produce a heterogeneous response, of predominantly indirect PFC within 7 days of transfer only when thymus cells are transferred along with the B cells. B cells from fetal or neonatal donors transferred without young adult thymus cells develop the capacity to generate indirect PFC within 13 days after transfer to adult recipients, but continue to produce a response of restricted heterogeneity of affinity for up to 28 days after transfer. Thus, it has been shown that cells present in the thymus facilitate, or are necessary for the functional maturation of B lymphocytes. Furthermore, the data suggest that maturation of the B-cell population to produce a heterogeneous response is controlled independently of its maturation to be capable of producing indirect PFC.


Alloantigen receptors on activated T cells in mice. I. Binding of alloantigens and anti-idiotypic antibodies to the same receptors. B6 alloantigens in supernates from one-way mixed lymphocyte reaction (MLR) cultures of AKR T cells against B6 lymph node cells rebound specifically to (B6)AKR-T-cell blasts after overnight incubation and recovery of these blasts from trypsin treatment. A similar specificity was observed with the binding of SJL alloantigens to (SJL)AKR-T-cell blasts. In both cases, the corresponding alloantigens were rebound several-fold more efficiently than control alloantigens. In a different assay system, T-cell receptors were studied with anti-idiotypic sera. These antisera were raised by repeated injection of purified (B6)AKR-T-cell blasts into (AKR X B6)F1 hybrid mice. These F1a(AKRaB6) sera reacted with the majority of (B6)AKR-T and (B6)(AKR X SJL)F1-T-cell blasts. They also reacted with a lower, though sizable, number of (SJL)AKR-T- and (SJL)(AKR X B6)F1-T-cell blasts. No reaction was observed with (B6)SJL-T, concanavalin A-activated AKR T-cell blasts, normal B6, (AKR X B6)F1, and (AKR X SJL)F1 T cells. Normal AKR T cells were positive only minimally above background. F1a(AKRaB6) sera could be made specific for (B6)AKR-T and (B6)(AKR X SJL)F1-T-cell blasts by absorption of contaminating antibodies with (SJL)AKR-T-cell blasts. Finally, it was shown by competition experiments that the receptors on MLR-activated T-cell blasts that bind alloantigens were the same as those binding anti-idiotypic antibodies. In addition, it was found that at least a fraction of alloantibodies share common idiotypic determinants with receptors on MLR-activated T-cell blasts.


Induction of specific immune unresponsiveness with purified mixed leukocyte culture-activated T lymphoblasts as autoimmunogen. II. An analysis of the effects measured at the cellular and serological levels. T lymphoblasts specific for foreign histocompatibility antigens and purified via mixed leukocyte culture (MLC) and 1 g velocity sedimentation procedures can be used as autoimmunogen to produce specific immunological unresponsiveness in adult animals. This unresponsiveness is positively correlated to the production of autoanti-idiotypic antibodies in the blast immunized animals and no evidence of coexisting alloimmunity was found. We consider this autoanti-idiotypic immunity to be the specific inducing agent of the immune tolerance. The blast immunization procedure will lead to selective reduction in T-cell reactivity against the relevant alloantigens as measured by MLC, cell-mediated lympholysis, or graft-versus-host assays. However, in individual animals, dichtomy in suppression between two T-cell assays could sometimes be observed indicating elimination of only a select group of idiotypic functionally distinct population of T cells in these blast-immunized animals. Attempts to abrogate already immune animals by the autoblast procedure were successful, in part suggesting the use of the present procedure when trying to induce in accelerated reversion of such immunity.


Induction of specific immune unresponsiveness with purified mixed leukocyte culture-activated T lymphoblasts as autoimmunogen. III. Proof for the existence of autoanti-idiotypic killer T cells and transfer of suppression to normal syngeneic recipients by T or B lymphocytes. Specific immune unresponsiveness against a given set of histocompatibility antigens can be induced by immunization with autologous, antigen-specific T lymphoblasts. Such unresponsiveness can be transferred by lymphoid cells from autoblast-immunized donors to normal syngeneic recipients. The cells being most efficient in transferring the selective suppression are T lymphocytes from the spleen, especially if of Ly 1-2+3+ phenotype. By using such T lymphocytes we deem it likely that the actual underlying mechanism is one of actual transfer of autoanti-idiotypic killer T cells. In support for this view is the fact that such T cells endowed with exquisite specific, cytolytic reactivity towards autologous idiotype-positive T target cells exist in autoblast immune animals. Significant suppression may also be transferred with T cells of Ly 1+2-3- phenotype or with B cells. Here, we consider the suppressive mechanism to be one of production of autoanti-idiotypic antibodies. By using affinity fraction procedures, it was finally possible to prove that all T-cell suppressive activity resides in a population with true antigen-binding-specific receptors for the relevant idiotypes.


H-2-restricted cytotoxic effectors generated in vitro by the addition of trinitrophenyl-conjugated soluble proteins. Murine spleen cells from normal donors were cultured in vitro with trinitrobenzene sulfonate (TNBS)-conjugated soluble proteins, i.e., bovine gamma globulin (TNP-BGG) or bovine serum albumin (TNP-BSA). Addition of 100 mug of any of these TNP-proteins to the spleen cell cultures led to the generation of cytotoxic T-cell effectors which were H-2-restricted and TNP- specific. The lytic potential of such effectors was comparable to that generated by sensitization with TNBS-modified syngeneic cells, and was restricted to haplotypes shared at the K or K plus I-A, or the D regions of the H-2 complex. Greater effecter cell activity was generated by addition of TNP-BGG against TNBS-modified targets which shared K plus I-A than against modified targets which shared the D region with the responding cells, which suggests that the same immune response genes are involved when the response is generated by the addition of TNP-conjugated soluble proteins or of TNBS- modified cells. H-2-restricted, TNP-specific effecter cells were generated by culturing mouse spleen cells with syngeneic cells which had been preincubated with TNP- BGG or TNP-BSA for 1.5 h. The addition of unconjugated soluble proteins to the cultures did not result in cytotoxic effectors detectable on H-2-matched targets, whether the targets were prepared by modification with TNBS, or by incubation with either the unconjugated or TNP-conjugated proteins. Depletion of phagocytic cells in the tumor preparation by Sephadex G-10 column fractionation before incubation with TNP-BSA had no effect on their lysis by the relevant effector cells. Immunofluorescent staining of tumor target cells with anti-TNP antibodies indicated that TNP could be detected on the tumor cells within 10 rain of incubation with TNP-BSA. The cytotoxic response generated by addition of the TNP-proteins to spleen cell cultures was found to be T-cell dependent at the effector phase, as shown by the sensitivity of the lytic phase to absorbed RAMB and complement. Furthermore, the response did not appear to be attributable to antibody-dependent cellular cytotoxicity. Three mechanisms were considered which could account for the generation of H-2-restricted, TNP-specific, cytotoxic T-cell effectors by the addition of soluble TNP-proteins. These include covalent linkage of activated TNP groups from the soluble proteins to cell surface components, macrophage processing of the soluble conjugates and presentation to the responding lymphocytes in association with H-2-coded self structures, or hydrophobic interaction of the TNP-proteins to cell surfaces. Results obtained from sodium dodecyl sulfate gel patterns indicating that cell-bound TNP was still linked to BSA, and the observation that phagocytic-depleted cells could interact with the soluble TNP-proteins and function as H-2-restricted targets, appear not to favor the first two proposed mechanisms.


Demonstration and quantitation of activation of the first component of complement in human serum. Activation of the first component of human complement (C1) in human sera can be readily detected in double immunodiffusion studies with anti-C1q, anti- C1r, and anti-C1s as it produces a characteristic pattern quite different from that of precursor C1. Native macromolecular C1 gives a continuous line of precipitation with antisera to C1q, C1r, and C1s in double diffusion studies. After activation of C1 by incubation of serum with complement activators, three major changes occurred in the Ouchterlony pattern. First, spurring of the C1s precipitin line over that of macromolecular C1, indicating release of C1s from C1, was observed with low doses of activator. Release of C1s was quantitated by single radial diffusion and shown to be complete with the highest activator dose examined. Second, C1q was released with larger activator doses as shown also by spurring of the precipitin line due to this component over the remaining macromolecular C1. Third, and most surprising, C1r antigenicity was progressively lost as the activator dose was increased and no C1r line remained with the highest dose of activator tested. This was not true with C1s as there was no change in the total C1s concentration in serum incubated with various activator doses. These observations provide two approaches to the quantitation of C1 activation in human serum. First, C1r and C1s can be quantitated by single radial diffusion. A decrease in the C1r:C1s ratio correlates with activation. Second, C1s released by the activation can be quantitated by single radial diffusion if the agarose contains high concentrations of anti-C1q to confine C1, also containing C1s, to the area near the application well, and lesser concentrations of anti-C1s to permit free C1s to produce a measurable ring. The extent of release of C1s also correlates with activation. These immunochemical techniques to quantitate C1 activation directly inserum do not require specialized reagents. It is hoped that they will be useful in screening pathological sera and in monitoring the status of the complement system in patients.


Increased synthesis and expression of H-2 antigens on thymocytes as a result of radiation leukemia virus infection: a possible mechanism for H-2 linked control of virus-induced neoplasia. Previous studies from this laboratory have mapped resistance and/or susceptibility to radiation-induced leukemia virus (RadLV)-induced neoplasia to the H-2D region. H-2 linked effects on virus replication can be detected subsequent to the initial virus infection, and clear-cut differences in numbers of virus infected thymus cells can be detected as early as 5 wk after RadLV inoculation. Rapid increases in cellular synthesis and cell surface expression of H-2 antigens are detectable immediately after virus inoculation. These changes have been studied by immunofluorescence, absorption, cell surface iodination followed by sodium dodecyl-sulfate-polyacrylamide gel electrophoresis, and two dimensional gel electrophoretic analysis of internally labeled lymphocyte proteins. Expression of H-2K molecules is significantly increased in cells of susceptible and resistant animals. However, significant increases in expression of H-2D antigens occurs only on thymus cells from resistant strains (H-2Dd). Transformed cells of resistant and susceptible H-2 haplotypes adapted to tissue culture lack detectable H-2 antigens as determined by serological absorption studies. It is argued that altered expression of H-2 antigens plays a very significant role in the mechanism of host defense to virus infection.


Characterization of human high molecular weight kininogen. Procoagulant activity associated with the light chain of kinin-free high molecular weight kininogen. Human high molecular weight (HMW) kininogen has been isolated and was found to be a single chain protein of approximately equal to 120,000 daltons. Upon digestion with plasma kallikrein bradykinin is generated, and SDS gel electrophoresis of the kinin-free protein reveals an apparent loss in size of 15,000 daltons. The kinin-free kininogen retains full activity as a coagulation factor and consists of two chains: a heavy chain of approximately equal to 66,000 daltons disulfide-linked to a light chain of 37,000 daltons. The heavy chain of HMW kininogen shares antigenic determinants with LMW kininogen and possesses no detectable coagulant activity. The isolated light chain is shown to be responsible for the coagulant activity of HMW kininogen and contains a unique antigenic determinant that distinguishes HMW kininogen from low molecular weight kininogen.


Human migration inhibitory factor: purification and immunochemical characterization. Using gel filtration and preparative isotachophoresis, the migration inhibitory factor (MIF) was highly purified from human lymphocytes activated with concanavalin A. MIF is an acidic protein with a mol wt of approximately equal to 25,000 daltons as determined by gel filtration and analytical polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The protein inhibits migration of macrophages in the capillary test and in addition, has a slowing effect on the electrophoretic mobility of guinea pig peritoneal macrophages. Rabbit antibodies specific for this protein, as determined by immunochemical techniques, neutralized the biological effect of MIF on migration and on the electrophoretic mobility of macrophages.


The role of H-2 linked genes in helper T-cell function. II. Isolation on antigen-pulsed macrophages of two separate populations of F1 helper T cells each specific for antigen and one set of parental H-2 products. A method was established for isolating antigen-specific murine helper T cells by selective binding to antigen-pulsed macrophage (Mphi) monolayers. Sheep erythrocyte (SRBC)-primed T cells, which remained strongly adherent to SRBC-pulsed syngeneic Mphi after 20 h in culture, were markedly enriched for helper activity when tested in the in vitro antitrinitrophenol (TNP) response to TNP-SRBC. Successful binding and enrichment occurred only if the Mphi were pulsed with the specific antigen to which the T-cell donors had been primed. The genetic control governing helper function in this system was then examined by using primed F1 T cells isolated on Mphi monolayers from congenic strains bearing parental H-2 haplotypes. SRBC-primed BDF1 (H-2b X H-2d) T cells, which bound to SRBC-pulsed H-2d Mphi, subsequently functioned as helper cells in cultures containing H-2d B cells and Mphi, but not in those containing H-2b B cells and Mphi. They remained unable to collaborate with B cells of the H-2B haplotype even in the presence of additional H-2d Mphi, indicating that H-2 restriction occurs at least at the level of the B cell. Similary, primed BDF1 T cells isolated on H-2b Mphi cooperated preferentially with H-2b B cells and Mphi. In both cases, the haplotype preference of the T cell was not due to alloreactive suppressor activity. These results suggest that primed F1 mice contain individual populations of helper T cells, each of which recognize antigen in association with a parental H-2 gene product(s) expressed during both Mphi-T cell and T cell-B cell interactions.


Learning modules as a supplement to existing audiovisual aids. The development and use of supplementary learning modules for audiovisual aids designed for wider audiences was found to be a feasible approach for the education of family practice residents. This approach allows programs to direct learner attention to areas of greatest interest in family practice. It was not excessively time consuming to develop the two modules and extensive technical expertise with media was not required. It was found that the modules were adaptable to individual or group use. This teaching method appears to be an excellent learning tool for residents. Supplementary learning modules may be useful for medical students and other health-care providers in family practice. Further economy of time and other innovations using this basic approach could result from recruiting family practice residents and other students to develop their own learning modules on topics of individual interest.


Suppression of murine leukaemia virus production by ouabain and interferon in mouse cells. Ouabain markedly inhibited the growth of the mouse cell lines K3b and JLS-V9 and the production of murine leukaemia virus (MuLV) in them. The inhibition of MuLV production was abolished by exposing the cells to normal medium or by adding a high concentration (43 mM) of K+ ions to the ouabain-containing medium. MuLV production was reduced by ouabain more rapidly than host cell directed protein synthesis. After treatment of cells with ouabain (0.5 mM) for 7 h, extracellular reverse transcriptase reverse transcriptase activities were reduced by 87 to 92%. However, the intracellular level of polymerase activities remained almost unchanged (77 to 98% relative to the control). Mouse interferon inhibited the production of MuLV in K3b cells and this antiviral action was not blocked by 0.5 mM-ouabain.


The specific reduction of a and d antigenic determinants after periodate treatment of hepatitis B surface antigen (HBsAg). HBsAg antigenicity was found to be sensitive to periodate treatment. Antigenic determinants a and d were especially sensitive, losing almost all of their activity, while determinants r and w were found to be quite stable. Also, it is now possible to prepare the monospecific antibodies, anti-r and anti-w, by the use of this new, simpler procedure.


Infection of human cell cultures with bovine visna virus. Fibroblastoid cell cultures derived from leukaemic bone marrow were successfully infected with BVV. After 2 months of subcultivation the cultures showed the appearance of foci of altered cells, suggestive of malignant transformation. Such foci were absent in non-inoculated cultures. Both control and inoculated cultures had a limited life span, i.e. neither of them could be developed into continuous transformed cell lines. The presence of at least some BVV genome functions in the inoculated cells was demonstrated (i) by immunofluorescence using a reference BVV serum, (ii) by detection in the supernatant culture fluid of sedimentable particles bearing RNA-dependent DNA polymerase activity with preference for Mg2+ ions, and (iii) by electron microscopic detection of scarce cell-associated virus particles in one of the infected cultures. Infectious BVV could not be rescued. In contrast to leukaemic bone marrow cultures, diploid human embryonic fibroblasts of various origin could not be infected with BVV.


Influence of transcutaneous nerve stimulation (TNS) on acute pain. Using transcutaneous nerve stimulation (TNS) simple surgical procedures such as tooth extractions and nerve biopsies can be performed without the usual anesthetics. Estimation of threshold and suprathreshold intensities of painful electrical stimuli show no significant change during TNS. Only the threshold for non-painful electrical stimuli is slightly increased. Cortical potentials evoked by electrical peripheral nerve stimulation are not significantly modulated by TNS. Latencies of the early components 0, I--III are unchanged, the amplitudes only slightly reduced. These observations are in contradiction to the 'gate-control' theory of pain.


Relationship between arteriosclerosis and cerebral atrophy in Parkinson's disease. Computed tomographic examinations of parkinsonian patients revealed a high incidence of cerebral atrophy, in most cases a combination of cortical atrophy and ventricular enlargement. The present study considered the relationship between cerebral atrophy and physical signs indicating or promoting arteriosclerosis such as overweight, electrocardiographic changes, hypertension, calcification of the internal carotid artery and aorta as well as elongation of the aorta. The study is based on 173 treated and untreated parkinsonian patients (89 men, 84 women) aged from 37--84 years (mean 64.6), on whom CT was performed about 5.4 years after the onset of the first symptoms of the illness. The results demonstrate an increase of pathological CT findings as well as of calcification in the carotid siphon with advanced age. No correlation was found between the other items and increasing age. Further analysis of the relationship between cerebral atrophy and signs of arteriosclerosis revealed only a statistically relevant correlation with calcification of the carotid siphon, especially with calcification of the media. Since pathological CT findings and calcification of the internal carotid artery are both related to advanced age, whereas all the other items which may be considered to be indications of arteriosclerosis do not have any clear relationship, it is concluded that the cerebral atrophy in Parkinson's disease is not caused by arteriosclerosis.


Electrophysiological recovery after compression ischemia of the rat brain. Cerebral ischemia of 15 min was produced in rats by increasing intracranial pressure above the systolic blood pressure level by infusing artificial cerebrospinal fluid into the cisterna magna. Pulmonary edema was prevented by blocking the vasomotor response with a ganglioplegic agent, and reducing the fluid load by peritoneal dialysis. The EEG flattened after 10.1 +/- 1.5 sec. followed by a dilatation of the pupils after 1--2 min. The EEG began to recover in 21 of 32 rats after 8 to 35 min of recirculation. Spectral analysis revealed an initial return of slow waves and spindles followed by continuous fast frequency activity. The EEG did not recover or was secondarily suppressed in 12 animals after a few hours. Electrophysiological recovery depended on the cardiocirculatory and respiratory state, the recovery being optimal in animals with mild hypertension and a normal acid-base status of the blood.


A case of demyelinating encephalomyelitis with some resemblance to collagen disease. A woman, aged 26 years, who died of progressively worsening demyelinating encephalomyelitis in the course of 4 years is reported. The neuropathological findings included large subcortical softenings in the cerebral hemispheres, tiny perivenous demyelinated foci in their neighborhood and scattered in the white matter. There was an acute vasculitis with fibrinoid exudation in the affected as well as unaffected areas. Focal perivenous mononuclear cell infiltrations are conspicuous in the white matter. The laboratory and postmortem examinations suggested a collagen disease like SLE. The abnormalities included marked increase of serum gamma-globulin, especially of IgG, and elevation of CRP, RA, and ANA titer, moderate thickening of the basement membranes of the renal glomeruli, onion skin-like periarteriolar fibrosis in the spleen, fibrous pericarditis and periadventitial fibrosis of myocardial arteries. Bilateral degeneration of the spinal posterior columns and dorsal roots was also observed. A probable relationship of the modified features in this example of demyelinating encephalomyelitis with abnormal immune mechanisms in the background is discussed.


[Age, duration of disease and survival period of patients with involvement of the nervous system by malignant lymphomas. A statistical comparison of mycosis fungoides with lymphogranulomatosis (author's transl)]. A statistical comparison dealing with age, duration of disease and survival period of patients with involvement of the nervous system by malignant lymphomas revealed the following data: 1. With mycosis fungoides, involvement of the nervous system occurs later in life than with lymphogranulomatosis; this difference is statistically highly significant (P less than 0.001). 2. There is no statistical difference (P greater than 0.05) in the duration of either type of malignant lymphoma before the nervous system becomes involved. 3. It is of high statistical significance (P less than 0.001) that, once the nervous system has become involved, the survival period is lower with mycosis fungoides than with lymphogranulomatosis. It can be expected that involvement of the nervous system by mycosis fungoides leads with 79% probability (confidential limits 99% = 47.29--96.22%) to death within 6 months.


Bilateral symmetrical softening of the thalamus. The clinical and pathological data of a 48 year old patient who survived 40 days are reported. Her disturbance of consciousness corresponded to an apallic syndrome, which 12 days later bacame akinetic mutism. Symmetrical softening involving the medial thalamic nuclei from the plane of the corpora mamillaria to the red nuclei was found. The ischemic lesion might be explained by transient circulatory collapse combined with hypoplasia of the vertebrobasialr arteries. On the EEG slight irregular alpha activity was recorded (alpha coma) and external stimuli elicited theta-delta waves (paradox activation). A survey of the literature of akinetic mutism is included and the correlation between non-hypnoid unconsciousness and EEG is discussed.


The anterior tarsal tunnel syndrome.

The anterior tarsal tunnel syndrome. The anterior tarsal tunnel syndrome, first described in 1968 by Marinacci, is characterized by a compression of the deep peroneal nerve under the inferior extensor retinaculum. The patients complaint of pains on the dorsum of the foot, especially at night. Clinically result sensory deficits in the involved area between the first and second toes as well as paresis and atrophy of the extensor digitorum brevis. The distal latency of the deep peroneal nerve is increased, the EMG shows active and chronic denervation of the extensor digitorum brevis. In cases with partial anterior tarsal tunnel syndrome only the motoric branch to the extensor digitorum brevis or only the sensory branch of the deep peroneal nerve after the division under the inferior extensor retinaculum is compressed. Two cases with complete and one with partial anterior tarsal tunnel syndrome are presented, etiology, symptomatology, differential diagnosis and therapeutic possibilities are discussed.


Crossed immunoelectrofocusing of cerebrospinal fluid immunoglobulins. The combination of the high separation capacity of isoelectric focusing and the specificity of immunoelectrophoresis gives a valuable tool for protein studies. This technic can be applied to immunoglobulin examinations by using KOCN-treated antibodies isoelectric at pH 5 and performing the immunoelectrophoretic procedure at this pH. CSF and serum samples of patients exhibiting CSF immunoglobulin abnormalities and blood-CSF barrier damages were examined by this method.


Electrophysiological studies on the "stiff-man" syndrome. In addition to anamnestic and clinical data electrophysiological and pharmacokinetic investigations may be necessary for the diagnosis of stiff man syndrome. Continuous activity of motor units with superimposed bursts during muscular spasms was found by electromyography in the two patients reported. Rigidity and continuous activity disappears during sleep, after i.v. application of Tubocurarine and Diazepam, during Thiopenal anesthesia and after neural block with Procaine. Dipropylacetate and Baclofen improved the condition but did not lead to complete relaxation. Biperidenlactat and Magnesiumlaevulinat have only a temporary effect on rigidity. Neostigmine, Phenytoine, Glycine, Dopa and 5-Hydroxy-Tryptophan had no effect. Passive shortening or stretching of the m. biceps brachii as well as touching the skin increased motor activity which spread to other segments and to the contralateral side. The H/M ratio was increased but the silent period was normal. A combination of Diazepam and Dipropylacetate or Clonazepam was therapeutically effective in the cases reported. A central genesis, of the pathogenetic mechanisms discussed, is the most probable in our cases.


Effects of hemodialytic treatment on uremic polyneuropathy. A clinical and electrophysiological follow-up study. The involvement of peripheral motor and sensory nerve, at least on a subclinical level, is nearly constant event with chronic renal failure. The study of the motor and sensory propagation velocity indicates that a widespread functional lesion of the axon with a peripheral point of attack and secondary demyelination, may be the basic pathogenetic event of uremic polyneuropathy. Prolonged hemodialytic treatment is substantially unable to influence the evolution of uremic polyneuropathy. The electrophysiological follow-up study of the peripheral nerve does not seem to be an index of adequate dialysis.


Follow-up study of myasthenic children after thymectomy. The results of thymectomy of 28 myasthenic children observed from 1 to 9 years after operation are discussed. Improvement was obtained in 19 cases (67.8%), with full remission in 12 cases (42.8%). Six children died. No correlation was found between the results of thymectomy and the duration of the disease, sex or histological findings in the thymus. No increase of incidence or severity of infections were observed in thymectomized children. Caution is recommended regarding vaccination of myasthenic children.


The influence of local cooling on neuromuscular transmission in the myasthenic syndrome of Eaton and Lambert. The effect of local cooling was studied in two patients with the Eaton-Lambert syndrome. We stimulated the ulnar nerve and registered the compound muscle action potential, the twitch force, and the maximum tetanic force. A distinct improvement of neuromuscular transmission occurred after cooling. The amplitude of the muscle action potential and the twitch force were larger and the time of the postactivation facilitation was longer. This effect of cooling is similar to the effect of guanidine on neuromuscular transmission. The release of acetylcholine at the nerve terminal is probably enhanced by cooling.


Tumor-specific antigens on rat liver cells transformed in vitro by chemical carcinogens. With the use of membrane immunofluorescence and xenogeneic antisera, tumor-specific membrane antigens were detected on rat epithelial-like liver cells transformed in vitro by chemical carcinogens. These antigens were not detected in 10-, 15-, and 19-day rat fetuses. Xenogeneic antisera were produced in rabbits by immunization of the rabbits with cultivated BD rat liver cells transformed by dimethylnitrosamine or N-methyl-N'-nitro-N-nitrosoguanidine. The specific antisera against tumor-associated antigen(s) were obtained by in vivo absorption in syngeneic male rats and by in vitro absorption with various cell lines. One tumor-specific individual antigen and two tumor-specific cross-reacting antigens were shown to be present on the surface of chemically and/or spontaneously transformed rat liver cell lines. They were not detected on liver and spleen cells of normal BD adult rats, on fetal liver cells, or on liver and intestinal carcinoma cells of Wistar rats. Sera from multiparous pregnant rats had no antibodies against these tumor antigens (although they reacted with fetal cells).


Morphologic and cytochemical comparison of human lymphoblastoid T-cell and B-cell lines: light and electron microscopy. Human lymphoblastoid cell lines characterized as T- or B-cells by various markers were compared morphologically and cytochemically by light and electron microscopy. Distinct differences in nuclear morphology, amount of cytoplasm, pyroninophilia, and periodic acid-Schiff (PAS) staining enabled us to discriminate between T- and B-cell lines. T-cells had nuclei with an irregular configuration, stippled heterochromatin, and small or absent nucleoli. The scanty cytoplasm of T-cells contained intensely stained, PAS-positive globules and was less pyroninophilic than the cytoplasm of B-cells. B-cells had more rounded, uniform, vesicular nuclei with prominent nucleoli and peripheral heterochromatin. The cytoplasm of B-cells was abundant and strongly pyroninophilic. Transmission electron microscopy generally confirmed these morphologic differences. These findings supported our contention that consistent cytologic features concordant with immunologic markers make it possible to identify certain lymphomas as being of B- or T-cell origin on purely morphologic grounds.


In vitro alpha-fetoprotein synthesis by monkey hepatocellular carcinoma. Five monkeys were treated ip with N-nitrosodiethylamine (DENA), and one was treated with 1-nitrosopiperidine (PIP), starting within 2 months of birth, until hepatocellular carcinoma (HCC) developed. All animals except the PIP-treated monkey had much elevated serum alpha-fetoprotein (AFP) values. Fresh, minced, biopsy-derived tumor was cultured with L-[14C]leucine and L-[14C]lysine. Synthesis of AFP was determined by radioimmunoassay and by specifically precipitable [14C]AFP. Good agreement between these two parameters was obtained for the 4 DENA-induced tumors synthesizing AFP in culture. Tumor from 1 DENA-treated monkey did not synthesize AFP. In addition, neither normal liver nor tumor from the PIP-treated monkey showed AFP synthesis. Rates of synthesis were 0.37-5.50 ng AFP/mg tumor/day, or 0.0012-0.0183 pg AFP/cell/day (if one assumes 3.0 X 10(5) cells/mg tissue) over 48 or 72 hours. Different nodules from the same animal had similar rates of synthesis. For tumors that synthesized AFP in culture, a positive correlation was generally found between rate of synthesis and serum AFP level. The rate of in vitro AFP synthesis observed was lower than that of immunoglobulin synthesis in human myeloma or of AFP synthesis in a rat HCC, but it was close to the estimated rate of AFP synthesis in a monkey HCC line in long-term culture.


Leukocyte migration inhibition of tumor antigen and purified protein derivative reactivity in guinea pigs sensitized to line 10 hepatocarcinoma and BCG. Agarose microdroplet leukocyte migration inhibition (LMI) assays were performed to measure reactivity against line 10 hepatocarcinoma antigens and purified protein derivative (PPD) with the use of peripheral blood leukocytes from line 10 and/or BCG-sensitized syngeneic guinea pigs. The assay was quite sensitive and detected leukocyte migration inhibition with concentrations as low as 12.6 ng protein/ml of the crude sonicate of the line 10 tumor and 0.1 pg PPD. Specificity was shown by lack of reactivity in leukocytes of line 10 and/or BCG-sensitized animals with antigen preparations of L2C leukemia cells or normal syngeneic liver. Furthermore, leukocytes from normal control guinea pigs failed to react with any antigen. The results also suggested antigen cross-reactivity between line 10 tumor and BCG. Leukocytes from guinea pigs sensitized to only BCG became LMI reactive to the line 10 sonicate as well as PPD. No reactivity was observed with leukocytes of the animals in simultaneous tests with a sonicate of guinea pig L2C leukemia cells. The results demonstrated the usefulness of this microassay in detection of LMI reactivity with low antigen concentrations and small volumes of whole blood.


Reverse transcriptase of RNA tumor viruses. V. In vitro proteolysis of reverse transcriptase from avian myeloblastosis virus and isolation of a polypeptide manifesting only RNase H activity. Purified avian myeloblastosis virus reverse transcriptase contains two subunits that are structurally related. The large subunit, beta (molecular weight, 95,000), was converted in vitro by chymotrypsin into a polypeptide of molecular weight 63,000. This polypeptide was indistinguishable from the small subunit, alpha (molecular weight, 65,000), in its chromatographic behavior on the phosphocellulose column and its tryptic peptide composition. During this proteolytic conversion, a polypeptide of molecular weight 32,000 (fragment B) was obtained. It was composed of tryptic peptides unique to beta and appeared to be derived from the portion of the beta subunit that was cleaved off during the conversion of beta into alpha. Upon continued proteolysis, a smaller polypeptide of molecular weight 24,000 (fragment A) was generated. This polypeptide manifested only RNase H activity and shared common amino acid sequences with beta and alpha subunits. Fragment A did not share any amino acid sequence homology with fragment B.


Vagal preganglionic innervation of the cat heart--an attempt to identify the medullary "heart area" by the retrograde axoplasmic transport of horseradish peroxidase. The medullary "heart area", origins of the vagal preganglionic fibers innervating the heart, were identified in 19 cats with the use of horseradish peroxidase (HRP) as a retrograde axonal tracer. When HRP was injected in the A-V node area, S-A node area, right atrial wall and right ventricular wall, the labeled neurons were principally found in the lateral half of the dorsal motor nucleus (DMN) and in the medial solitary nucleus (MSN), were almost the same in number bilaterally, and were chiefly from the calamus scriptorius to the upper cervical cord. They showed no anatomical proof for asymmetric distribution. When HRP was injected in the left ventricular wall or the apex, few labeled neurons were identified in the DMN. Labeled neurons were not found in the nucleus ambiguus. This study suggests the labeled neurons in the DMN and MSN are origins of the dually innervating vagal preganglionic fibers to the heart.


Effects of passive antibody in bee venom anaphylaxis. Twelve patients allergic to honeybees were challenged by injections of bee venom; five responded systemically to the venom, with symptoms ranging from angioedema to respiratory distress. These patients were given intramuscular or intravenous infusions of gamma-globulin obtained from the plasma of hyperimmune beekeepers who had high levels of antibody to an allergen (phospholipase A) in the venom. Post-infusion, all five patients tolerated 1.5 to 5 times the venom dose that previously elicited adverse reactions. The quantity of passive IgG antibody infused did not impair the patient's own immune response to venom. These results represent the best available evidence for a direct role for IgG blocking antibodies in clinical protection against anaphylaxis occurring as a result of parenteral antigenic challenge as may be observed in penicillin and insect hypersensitivity.


[Detection of alpha2H-globulin in the blood serum in myocardial infarct]. The content of alpha2H-globulin in the plasma of 53 patients with macrofocal and 33 patients with microfocal myocardial infarction was determined in dynamics by the immunochemical method, with due account for the form and severity of the disease, the extent of involvement of the heart muscle, and the presence of complications and concomitant pathologic conditions of the lungs and kidneys. The frequency of alpha2H-globulin detection was 62% in macrofocal and 21% in microfocal myocardial infarction. In transmural myocardial infarction alpha2H-globulin is detected within the first 3 days of the disease in 37.5% of cases. The appearance of alpha2H-globulin in the blood plasma of patients with myocardial infarction is evidently associated with hypoxia and cell regeneration.


[Limited diffusion of dye between choroid and retina during animal experiments after injection of fluorescein and rhodamin (author's transl)]. For contributing to the question of an existence of a diffusion of the unbound part of the fluorescein between choroid and retina in fluorescein angiography, on pigmented rats, after intravenous injection of Fluorescein sodium and Rhodamine B respectively, comparing observations were effectuated. Rhodamine B in its unbound form showed a distinct diffusion between choroid and retina and coloured all retinal layers in the late phases whereas in the case of the Fluorescein sodium, only a poor diffusion was observable. The similar spectral bands of the primary and secondary fluorescences render difficult the observation of the behaviour of the fluorescein dye.


[Quantitative estimation of cationic leukocyte antigen (cla) and lysozyme in leukaemic leukocytes (author's transl)]. Cationic lysosomal proteins such as cationic leukocyte antigen (CLA) and lysozyme were estimated in leukocyte lysates of acute and chronic leukaemics using quantitative enzymatic and immunoprecipitation techniques. The studies demonstrate that both lysosomal conponents are markers of non-lymphatic leukaemias. Therefore the determination of CLA and lysozyme is valuable for the differential diagnosis of acute leukaemias. The intracellular cationic protein contents showed characteristic kinetic variations as showing during X-irradiation of the spleen in cases of chronic myelocytic leukaemia. White cells in leukocytosis could be distinguished from normals by remarkably low cationic protein contents in leukocyte lysates. The results are interpreted in the light of current results revealed by immunofluorescent analyses.


Cytochemical study of human lymphocytes stimulated by PHA in function of donor age. The 3H-AM binding reflects the structural changes involved in the cellular differentiation. This parameter was studied during blastic transformation of human lymphocytes, in relation to the age of the donor. Although they are individual variations, the 3H-AM binding is higher in the young group than in the aged subjects, as well as the blast transformation score. These results indicate that the weak lymphocyte response to the mitogen lectine (PHA) stimulation could be related to some age-induced structural alterations of the chromatin, resulting in an irreversible blockage in G1, at least in some of the T lymphocytes.


Post-thyroidectomy thyrotoxicosis.

Post-thyroidectomy thyrotoxicosis. 94 patients with postoperative recurrent hyperthyroidism were evaluated for duration of remission, goitre size, and response to radio-iodine (131I). 6 patients required 131I therapy within twelve months of operation--5 had large remnants because of inadequate surgery. 57% of patients relapsed within 5 years, but 16% relapsed after 20 years and 8% after more than 30 years. Estimated goitre weights ranged from 4 g to 65 g, and goitre size was unrelated to the duration of remission. All patients were treated with 131I. 23% of the patients became hypothyroid in the first postoperative year and 10% in the second year. The results indicate that postoperative thyrotoxicosis can recur decades after operation. Operation seems to sensitise the thyroid to the early effects of radiation by 131I.


Return of epidemic A1 (H1N1) influenza virus. Limited outbreaks of influenza were reported in the far east of the U.S.S.R. early in November, 1977, and a week or two later in cities in Siberia and the European parts of the U.S.S.R. The majority of influenza-virus isolates belonged to the H1N1 subtype of influenza-A virus, though strains of the current H3N2 subtype were also isolated. A moderate epidemic of H1N1 influenza spread throughout the country in the middle of December. Mainly children over 5 years and young adults were infected.


Should cholecystectomy be combined with vagotomy and pyloroplasty? Duodenal ulcers and gallstones, two of the commonest surgical conditions, affect respectively 10% of men and up to 20% of the population. Although many detailed studies of the treatment of these conditions have been conducted, there is no report of the results of surgery when the conditions coexist. 60 patients who had undergone vagotomy, pyloroplasty, and cholecystectomy were compared with age and sex matched controls who had undergone vagotomy and pyloroplasty alone or cholecystectomy alone. In the early postoperative period after the combined procedure there was a very high incidence of post-vagotomy diarrhoea (48.3%, P equal to 0.00013) and bile-reflux gastritis. The findings implicate bile-acids--their excretion and handling by the small intestine--in the aetiology of post-vagotomy diarrhoea. Where the conditions coexist truncal vagotomy and pyloroplasty should be avoided in the treatment of the duodenal ulcer because of the risk of post-vagotomy diarrhoea and bile-reflux gastritis.


Subtraction potential difference: In-vivo assay for mineralocorticoid activity. Parallel fluctuations of potential difference (P.D.) across oral and rectal mucosa are probably related to the activity of autonomic nerves rather than adrenal steroids. Unlike rectal mucosa, oral mucosa does not respond to exogenous or endogenous aldosterone. Therefore subtraction of oral P.D. from rectal P.D. gives a closer indication of mineralocorticoid activity than does rectal P.D. alone. In normal subjects plasma-aldosterone correlated well with subtraction P.D. (r = 0.74; P is less than 0.001). A subtraction P.D. higher than 26 mV in subjects on a normal sodium intake indicated primary or secondary hyperaldosteronism; if the level was lower than 11 mV four hours after intramuscular injection of 0.25 mg tetracosactrin this suggested mineralocorticoid deficiency. Measurement of oral and rectal P.D. permits rapid and inexpensive diagnosis of aldosterone excess and deficiency. The method may also be used in study of the mineralocorticoid effect of other adrenal steroids: as assessed with this bioassay, the plasma 18-OH-deoxycorticosterone, which is raised in some patients with essential hypertension, lacked any in vivo mineralocorticoid activity.


Light-microscopical diagnosis of rabies. A reappraisal. The validity of reports suggesting that Lyssa and Negri bodies are non-specific in the light-microscopical diagnosis of rabies was investigated. With Seller's impression technique, a substantial proportion of speciments found to be non-rabid with the fluorescent-antibody technique showed structures indistinguishable from Lyssa or Negri bodies. Neither histological examination nor inoculation of animals with non-rabid Seller-positive material explained the nature of these rabies-like structures. The simplicity and reliability of the fluorescent-antibody technique and the occasional serious complications of prophylactic anti-rabies measures make the diagnostic use of Seller's method at best undesirable and at worst dangerous.


Common manifestations of cow's milk allergy in children. Cow's milk allergy was diagnosed in 79 patients, all of whom had signs and symptoms of allergies other than milk intolerance. In addition to difficulties with infant feeding and diarrhoea, clinical features included constipation, vomiting, intestinal colic, growth retardation, and psychological disturbance, as well as eczema and asthma. All were reversible after milk withdrawal, which suggests that the allergic basis of such symptoms may have been underestimated. In most cases, one or both parents were atopic and the child had been bottle-fed from birth. There were no breast-fed children of non-atopic parents in this series.


In-vitro synthesis of anti-acetylcholine-receptor antibody by thymic lymphocytes in myasthenia gravis. Cultures of thymic tissue or thymic lymphocytes from nine patients undergoing thymectomy for myasthenia gravis were examined for evidence of anti-acetylcholine-receptor-antibody synthesis. One of four thymic tissues and three of five thymic lymphocyte preparations synthesised this antibody.


Vagotomy for treatment of severe obesity. Three women weighing 137, 133, and 135 kg underwent truncal vagotomy; after 16, 20, and 24 weeks they had lost 10, 17, and 19 kg respectively without serious side-effects. No dietary restrictions have been imposed, but they are eating less than they were preoperatively and report a total lack of hunger. Truncal vagotomy may be a useful treatment for severe obesity.


Hyperinsulinism, diabetes mellitus, and respiratory distress of the newborn: A common link? Hyperinsulinism may reduce glycerol-3-phosphate and dihydroxyacetone phosphate production and thus impair phospholipid synthesis and surfactant production in the lung. This mechanism could be responsible for the increased incidence of respiratory-distress syndrome in the infants of diabetic mothers.


Respiratory-cancer clustering associated with localised industrial air pollution. Between 1968 and 1974 there was a significantly large number of deaths from respiratory cancer among members of a Scottish community living near and downwind from a steel foundry. The air in that area was polluted by metallic particles from the foundry.


Trial of prostaglandin-synthetase inhibitors in primary dysmenorrhoea. The prostaglandin-synthetase inhibitors, mefenamic acid and flufenamic acid, were compared with the analgesic, dextropropoxyphene/paracetamol, in the treatment of primary dysmenorrhoea in a double-blind crossover trial. Results were assessed in 30 patients who took each drug during menstruation for three consecutive cycles. The patients' assessment of each drug suggests that both mefenamic acid and flufenamic acid are more effective than the other analgesic for general relief of symptoms and for most of nine individual symptoms subjectively assessed by the patient. There was less absenteeism from work or school during mefenamic-acid treatment and fewer capsules of mefenamic acid were taken compared with the other two drugs. Patients took significantly fewer additional analgesics during mefenamic-acid therapy than during treatment with the other two drugs. 5 patients had possible side-effects--3 patients on mefenamic acid and 2 on dextropropoxyphene/paracetamol.


Flufenamic acid in treatment of primary spasmodic dysmenorrhoea. A double-blind crossover study. A double-blind cross-over trial of flufenamic acid three times a day (200 mg) was carried out in forty-four patients with primary dysmenorrhoea. While on flufenamic for 3 months 82% of patients experienced significant pain relief. Associated gastrointestinal symptoms, i.e--vomiting and diarrhoea--were relieved in 66% and 52% patients respectively while on flufenamic acid. It is concluded that the fenamates are useful and safe drugs in the treatment of primary dysmenorrhoea.


Contact with hospital, drugs, and chemicals as aetiological factors in leukaemia. 45 adults with acute leukaemia or chronic myeloid leukaemia and a control group of patients from the same hospital were asked about the people they had close social contact with before their illness, and about their use of drugs and chemicals. 18 (40%) leukaemia and 6 (13%) control patients had close social contact with hospital personnel or leukaemia patients. 8 (18%) leukaemia patients, but no control patients, had been in close contact with haemotological ward personnel. These differences were statistically significant. 9 (20%) leukaemia and 4 (9%) control patients lived in the same house as healthy persons working in a hospital. No conclusion could be drawn from differences between the two groups in their use of drugs, since the possibility that they were used to treat initial symptoms of leukaemia could not be excluded. Exposure to chemicals, including weed-killers and agricultural insecticides containing a benzene-ring known to be leukaemogenic, was about the same in the two groups.


Hycanthone dose-response in Schistosoma mansoni infection in Kenya. The recommended doses of the drugs now used to cure schistosomiasis mansoni may be associated with toxic side-effects. Since Schistosoma mansoni does not multiply in the human host and the disease seems to be closely associated with the intensity of infection, it may not be necessary to use 100% lethal antischistosomal doses, particularly in endemic areas. A dose-response to the antischistosomal drug, hycanthone was established for three different doses in 169 patients with heavy S. mansoni infections in the Machakos district of Kenya. The highest dose used (1.5 mg/kg or half the recommended package-insert dose) resulted in a 96% decrease in egg output (equivalent ot death of the worms); 0.75 mg/kg in an 85% decrease; and 0.375 mg/kg in an 11% decrease one month after treatment. In contrast to the vomiting common with the package-insert dose (3.0 mg/kg), there were no side-effects with any of the lower doses.


Association between treatment-resistant kidney-allograft rejection and post-transplant appearance of antibodies to donor B-lymphocyte alloantigens. In 18 kidney-allograft recipients the post-transplant appearance of antibodies to B-lymphocyte (DRw) alloantigens of the kidney donor was significantly associated with the rejection of the graft within 1 month of transplantation or the poor recovery of graft function, despite treatment for rejection. None of the 10 recipients whose grafts were accepted or whose renal function recovered well after treatment for rejection made antibodies to donor B-lymphocyte antigens after transplantation. DRw antigens are thus targets for the host's immune response to a graft and may be responsible for kidney rejection in patients who do not respond to rejection therapy.


Treatment of agoraphobia by subliminal and supraliminal exposure to phobic cine film. Three groups of 5 chronically agoraphobic patients were treated by repeated exposure to cine film at weekly intervals for six weeks. A supraliminal group saw an agoraphobic film specially made for the study illustrating a range of phobic situations. A subliminal group viewed the same film at a level of illumination below the threshold of awareness. A control group saw a blank screen with no filmed material for the same length of time. The films were screened five times at each treatment and were alternated with neutral film shown under normal viewing conditions. Ratings of phobic and other symptoms were made by a separate assessor who was unaware of the patient's treatment. The supraliminal and subliminal groups improved significantly more than the control group with regard to phobic fears, avoidance, and overall assessment.


Occlusion, carbon dioxide, and fungal skin infections. Occlusion of the skin renders it susceptible to acute fungal skin infections (dermatophytosis and candidiasis). Occlusion also raised carbon-dioxide (CO2) tensions at the skin's surface. Comparable CO2 tnesions have a pronounced effect on the morphology and metabolism of dermatophytes in vitro. It is postulated that dermatophyte conida and hyphae produce infective units under conditions of raised CO2 tensions, and that occlusion of the skin produces the concentrations of CO2 required for the conversion. Fungal skin infections might be prevented or controlled by interference with the action of CO2 or by prevention of its accumulation under wet, occlusive clothing.


A double-blind controlled crossover trial of an antigen-avoidance diet in atopic eczema. 20 out of 36 children (aged two to eight years) with atopic eczema completed a twelve-week, double-blind, controlled, crossover trial of an egg and cows' milk exclusion diet. During the first and third four-week periods, patients on an egg and cows' milk exclusion diet received a soya-based milk substitute (trial period) or an egg and cows' milk preparation (control period). Response was assessed in terms of eczema activity, number of areas affected, pruritus, sleeplessness, and antihistamine usage while on the two diets. During the middle period patients resumed their normal diet to minimise any carry-over effect. 14 patients responded more favourably to the antigen-avoidance diet than to the control diet, whereas only 1 responded more favourably to the control diet than the trial diet. Patients experienced more benefit during the first diet period than the second, whatever the nature of the diet. There was no correlation between a positive prick test to egg and cows' milk antigen and response to the trial diet.


Maintenance treatment of recurrent peptic ulcer by cimetidine. 68 patients with chronic peptic ulcer took part in a double-blind trial in which 32 received the histamine H2-receptor antagonist cimetidine, (400 mg twice daily) and 36 received a placebo, for a year. 6 of those on cimetidine had an endoscopically proven recurrent ulcer within a mean of 7 months, while 30 of those on placebo had one after a mean of 4 months (P less than 0.0005). 1 patient on cimetidine had two recurrences compared with 12 patients on placebo (P less than 0.0005). No patient in the cimetidine group had complications compared to 4 patients in the placebo group, 2 of whom had melaena and haematemesis and 2 melaena alone (P less than 0.05). 15 patients on placebo and 1 on cimetidine were referred for surgery during the trial because of symptoms from their recurrent ulcers. Day and night ulcer pain and antacid consumption were significantly less, and general wellbeing better, in cimetidine group. The only probable side-effect of cimetidine was reversible drug-induced liver damage of hypersensitivity type in 1 patient.


Beta-blockade and blood-levels after low-dose oral propranolol: The hepatic "first-pass" threshold revisited. Heart-rate, arterial pressure, and plasmarenin activity were determined in six normal subjects at rest and after an injection of 8 microgram isoprenaline with and without prior propranolol administered orally in a dose of 5 mg 8-hourly for a total of five doses. After propranolol, resting heart-rate, systolic pressure, and plasma-renin activity all fell significantly (P less than 0.05 to less than 0.001). When the isoprenaline-induced changes of heart-rate, diastolic pressure, and plasma renin activity without propranolol were compared to those with propranolol, these responses were greatly diminished (P less than 0.01 to less than 0.001). The percent blockade by propranolol of the isoprenaline-induced changes ranged from 65% for diastolic pressure to 77% for heart rate and 78% for plasma-renin activity. Propranolol levels determined by conventional fluorometry were below accurate detection limits, whereas those determined by gas-liquid chromatography ranged from 2.3 to 8.5 ng/ml. These findings, which demonstrate beta-blockade with low-dose propranolol, are not consistent with the existence of a postulated threshold for the hepatic "first-pass effect" in man, which is said to require saturation by single doses of 30 mg or more before propranolol enters the systemic circulation.


Legionnaires' disease in renal-transplant recipients. Legionnaires' disease is reported in five renal-transplant recipients. All had febrile respiratory illnesses with pulmonary infiltrates and one died. The diagnosis was made on clinical features and by indirect fluorescent antibody titres. Symptoms started after maximum immunosuppressive therapy.


Pulmonary oedema after radiological investigation of peripheral occlusive vascular disease. Adverse reaction to contrast media. 5 patients developed pulmonary oedema after retrograde femoral arteriography under general anesthesia. Because of the haemodynamic changes associated with radiographic contrast media, a good preoperative cardiological assessment is essential. The volume and nature of the contrast media injected and any other fluid administered should be carefully monitored. There is no apparent safe maximum dose of radiographic contrast media, but this work suggests that for 'Conray 420' (sodium iothalamate 70% w/v) a total dose should be less than 200 ml in a fit patient. The dose should be substantially smaller in patients with a history of evidence of myocardial infarction, myocardial insufficiency, myocardial ischaemia, or hypertension.


Sex of first child as a prognostic factor in breast cancer. The sex of the first child of patients who underwent mastectomy for potentially curable breast cancer appeared to be a valuable prognostic factor: patients whose first child was a boy had a better outcome than those whose first child was a girl. The difference was statistically significant. This may have been because the male/female sex ratio among first children was significantly higher in those patients without node involvement than in those with node involvement. But the favourable effect of a male first birth was still seen when only patients with an equal degree of node involvement (greater than or equal to 4 nodes) were studied. The "protection" resulting from a male first-born could be the result of fetal testicular secretions. This protection did not apply to the risk of breast cancer--the male/female sex ratio of first children in our series was 1.08, a figure not statistically different from that of the overall French population (1.05).


Plasmapheresis in Raynaud's disease.

Plasmapheresis in Raynaud's disease. Understanding of the pathogenesis, clinical course, and treatment of Raynaud's disease is confused and unsatisfactory, and no success in treatment can be promised. Five patients with severe Raynaud's disease, who had not responded to other forms of treatment, were treated by plasmapheresis, which produced a striking improvement. Ultrasonic velocimetry showed that segments of digital arteries which had been thought to be permanently occluded became patent and remained patent after plasmapheresis.


Disequilibrium in control of proteolysis as a cause of pre-eclampsia. The interrelated systems which control proteolysis may go into disequilibrium during pregnancy so that a renin-like enzyme in the plasma produces effects such as the early clinical manifestations of pre-eclampsia; the overactivity of the reninlike enzyme may in turn lead to undue activity of other physiological systems including those of the coagulation and complement systems.


"Food allergy": Fact or Fiction?

"Food allergy": Fact or Fiction? Six patients with longstanding physical and mental symptoms who had not been helped by many years of conventional medical investigation and treatment experienced immediate relief of symptoms when they avoided certain foodstuffs. This clinical study supports the view that some foods may cause widespread and disabling symptoms in people who are sensitive to them.


Susceptibility to diphtheria.

Susceptibility to diphtheria. Schick tests and antitoxin titrations have been carried out to investigate what percentage of individuals in each of 9 small groups were susceptible to diphtheria. Overall, 33% were susceptible: 26% among those below and 44% among those above 35 years of age. These results suggest that about 35% of the population of the United Kingdom are susceptible to diphtheria.


Respiratory hazards from papain.

Respiratory hazards from papain. Late-onset asthma occurred in a non-atopic worker in a factory where papain powder was packed. The patient had had 3 attacks of asthma since first being exposed to atmospheric papain dust. He gave a positive immediate reaction to skin-prick tests with solutions of papain which were appreciably weaker than solutions which caused no reaction in unexposed individuals. These results, together with his history and the fact that symptoms stopped when he was not exposed to papain, suggest that he had extrinsic allergic asthma caused by sensitisation to papain. Shortly after re-exposure to papain a worker in another factory died during an attack of asthma. Measures to prevent the inhalation of papain dust must be taken in factories where papain is handled, not only to avoid the proteolytic effects of the material but also to prevent workers from becoming sensitised.


Failure of the reaction of beta2-microglobulin with staphylococcal protein A. Because beta2-microglobulin is structurally similar to IgG, the reaction of beta2-microglobulin with Staphylococcal Protein A, which is known to react with the Fc region of IgG, was examined. 125I-beta2-microglobulin did not bind to Protein A. This may due to the difference in the amino acid sequence between beta2-microglobulin and the Fc region of IgG.


Phase II study of subcutaneously administered 5-azacytidine (NSC-102816) in patients with metastatic malignant melanoma. Thirty (30) patients with advanced metastatic malignant melanoma refractory to DTIC (NSC-45388) and a nitrosourea were treated with 5-azacytidine (NSC-102816). 5-Azacytidine was administered subcutaneously at a dosage of 100 mg/m2/day for 10 days. Twenty-six (26) patients were evaluable for toxicity and response. Major organ toxicities were hematologic, gastrointestinal, and cutaneous; no antitumor activity was noted.


Cytogenetic and other effects of the chemosterilants tepa, metepa, apholate an hempa in insects (a review). A review of the literature revealed that the chemosterilants tepa (tris(1-aziridinyl)phosphine oxide), metepa (tris(2-methyl-1-aziridinyl) phosphineoxide), apholate (2,2,4,4,6,6-hexakis(1-aziridinyl)-2,2,4,4,6,6-hexahydro-1,3,5,2,4,6-triazatriphosphorine), and hempa (hexamethylmelamine) affected both reproductive and somatic tissues in over 65 species of insects. The effects were cytological, physiological, and genetic and varied from slight to severe. In some cases the deleterious effects may have been species-specific, but in general, they appeared to be dose-dependent. More than 150 publications are cited.


The cytogenetic effect of bleomycin on human peripheral lymphocytes in vitro and in vivo. The cytogenetic effect of bleomycin (BLM) in human lymphocytes was studied after exposure to different doses during the G0 and G2 phases. BLM produced a marked specific effect on the cell cycle. The main aberration types after exposure in tg0 were dicentrics and deletions; and after exposure in G2, open chromatid breaks. A linear dose--response was calculated for all these aberration types as well as for the number of aberrant cells. In the G2 experiments, partially and totally pulverized cells also increased linearly with dose. The intercellular distributions of the most frequent aberration types after exposure in G0 and G2--the dicentrics and chromatid breaks, respectively--showed over-dispersion. These results show that the cytogenetic effect of BLM may be compared with that of densely ionizing irradiation. Preliminary results of chromosome analysis of three cancer patients in the course of BLM therapy showed effects similar to those in the G0 experiments.


Immunocytochemical identification of azurophilic and specific granule markers in the giant granules of Chediak-Higashi neutrophils. We used immunofluorescent microscopy to characterize the abnormal granules in neutrophils from five patients with Chediak-Higashi disease. Monospecific antiserums to the azurophilic markers myeloperoxidase, elastase, cathepsin G and lysozyme, and to the specific granule markers lactoferrin and lysozyme, were labeled with fluorescein and rhodamine and were used to demonstrate two antigens in the same cell simultaneously. The abnormal granules in Chediak-Higashi neutrophils contained both azurophilic and specific granule markers. Normal-appearing lactoferrin-positive granules were also present, but normal azurophilic granules were not seen. Analysis of bone-marrow samples from two of these patients suggested that the abnormal granules were formed during granulocyte maturation by the progressive aggregation and fusion of normally formed azurophilic and specific granules. These results are consistent with a membrane abnormality or a defect of microtubular function leading to inappropriate granule fusion, and suggest that the granular abnormality is more generalized than previously appreciated.


[Unusual clinical aspects of hyperthyroidism. Apathic hyperthryroidism]. The principal data in the literature regarding apathic hyperthyroidism are summarised and commented on 3 personal cases are then reported. The nosography of these forms is discussed and emphasis laid on pathogenetic hypotheses and diagnostic and therapeutic aspects. Diagnosis should be precise and early so as to prevent the extremely serious course which is a characteristic of this rare, but not exceptional variant of hyperthyroidism.


Antibodies to bacterial L-asparaginases. Complement fixation has been shown to occur when asparaginase reacts with specific antibody. This is as a result of an antigenic determinant common to the four bacterial asparaginases. Patients on asparaginase therapy may produce antibodies and these could be demonstrated by complement fixation.


Amniography in the prenatal diagnosis of neural tube defects. Amniography for the visualization and confirmation of suspected neural tube defect was performed in 9 midtrimester gravidas. In all cases, amniotic fluid alpha-feto protein (AFP) was abnormally elevated. Four cases of anencephaly and one of spina bifida were demonstrated by amniography. These pregnancies were terminated and the defects were confirmed by gross pathologic examination. In 4 remaining cases, amniography was normal. Three of these pregnancies proceeded to term, culminating in the birth of a normal child. The fourth patient had spontaneous abortion of a normal fetus at 23 weeks of gestation. The experience reported here suggests that amniography is an important adjunctive diagnostic technique in the prenatal diagnosis of neural tube defect, and if used correctly, may significantly reduce the chance of false-positive diagnosis.


Physiologic jaundice of the newborn. Relation to maternal serum and amniotic fluid alpha-fetoprotein. Maternal serum and amniotic fluid alpha-fetoprotein determinations were made at or near term. There appears to be a correlation between elevated levels of alpha-fetoprotein and physiologic jaundice of the newborn. It is suggested that this assay might be used as an index of fetal liver maturity when considering elective cesarean section or induction.


Comparison and evaluation of fluorescent antibody techniques in the detection of herpes simplex virus in oral infection. This article deals with the reliability and applicability of fluoroscopy in the diagnosis of herpetic lesions of the oral cavity. The reported results are based on a clinical experiment performed on fifty-two lesions suspected to be of herpetic origin. Three different methods were compared for detection of the possible presence of the virus--cytologic, virologic, and immunofluorescent. The greater reliability and speed of the last technique are demonstrated by our findings. Such features acquire special importance in the differential diagnosis of a number of diseases.


Brief screening for developmentally delayed preschoolers. 23 preschool boys and girls with developmental delays were administered the Peabody-A, Riley Design, McCarthy Designs, and the Riley human figure. Correlations among scores ranged from .58 to .80 suggesting preliminary screening might be undertaken by a qualified teacher though this sample was very small and special.


Selection of patients for coronary bypass surgery. We have tried to present a rational approach to selection of patients for coronary bypass surgery. An evaluation of both the anatomic and the clinical status of a patient in whom coronary arteriography has been performed, along with consideration of known results of medical and surgical forms of treatment of coronary artery disease in large series, determines which therapy is preferable. In all instances, the goal of treatment is threefold: relief of pain, preservation of myocardium and prolongation of life. In properly selected patients, pain can be relieved by surgery 90% of the time. Preservation of myocardium is a reality in some groups of patients treated surgically. Current data indicate that prolongation of life by coronary bypass surgery occurs in patients with left main coronary or triple-vessel disease, and as more data are collected, other subgroups may be included.


Radiotherapy for bronchogenic carcinoma. Treatment of carcinoma of the lung to cure or even to prolong survival time to a satisfactory degree is not yet a reality. For squamous cell carcinoma confined to the lung, radical surgery offers the best possibility of cure. When surgery is not feasible in such a case for other than technical reasons (eg, poor general condition of the patient), radical (megavoltage) radiotherapy offers a good possibility of cure. For anaplastic or oat cell carcinoma confined to the lung, cure by surgery or radiotherapy is less likely and there is little advantage of one or the other of these modalities. Once metastasis has occurred, surgery is largely precluded and radiotherapy becomes a measure of last resort that offers a good possibility of relief of distressing symptoms and some hope of prolonging worthwhile life.


Chemotherapy for lung cancer.

Chemotherapy for lung cancer. Oat cell carcinoma is the one variety of primary lung cancer which is sufficiently responsive to a variety of drugs and drug combinations that a very high rate of response and significant prolongation of survival can be consistently achieved. The question of whether anyone has been "cured" by drug treatment awaits more study and the passage of time. Progress in the treatment of the other varieties of bronchogenic carcinoma is less encouraging. There is only the hope that more fundamental knowledge about the nature of the disease will bring greater progress.


Whipple's disease in a female with impaired cell-mediated immunity unresponsive to co-trimoxazole and levamisole therapy. A case of Whipple's disease in a female is described. Malabsorption of iron, vitamin B12, folic acid and fat was present. These abnormalities reverted to normal after introduction of co-trimoxazole therapy. The patient's humoral immune system was normal, in contrast to impairment of cell-mediated immunity, which has not improved in spite of co-trimoxazole treatment for 2 years, and a therapeutic trial of levamisole. These findings are further evidence that there may be a primary immune deficiency in patients with Whipple's disease.


Inhibition of Rous sarcoma virus replication and cell transformation by a specific oligodeoxynucleotide. The tridecamer d(A-A-T-G-G-T-A-A-A-A-T-G-G), which is complementary to 13 nucleotides of the 3'- and 5'-reiterated terminal sequences of Rous sarcoma virus 35S RNA, was added to chick embryo fibroblast tissue cultures infected with Rous sarcoma virus. Inhibition of virus production resulted. The inference emerges that the tridecamer and its counterpart with blocked 3'- and 5'-hydroxyl termini enter the chick fibroblast cells, hybridize with the terminal reiterated sequences at the 3' and 5' ends of the 35S RNA, and interfere with one or more steps involved in viral production and cell transformation. Likely sites of action are (i) the circularization step of the proviral DNA intermediate, and (ii) the initiation of translation, the latter being described in the following communication [Stephenson, M. L. & Zamecnik, P. C. (1978) Proc. Natl. Acad. Sci. USA 75, 285--288].


Inhibition of Rous sarcoma viral RNA translation by a specific oligodeoxyribonucleotide. A tridecamer oligodeoxynucleotide, d(A-A-T-G-G-T-A-A-A-A-T-G-G), which is complementary to reiterated 3'- and 5'-terminal nucleotides of Rous sarcoma virus 35S RNA, is an efficient inhibitor of the translation of proteins specified by the viral RNA in the wheat embryo cell-free system. The inhibition specificity for oncornavirus RNA is greater than for rabbit reticulocyte mRNA or brome mosaic virus RNA. Other oligodeoxynucleotides of similar size have little or no specific effect on the RNA-directed translation. The tridecamer acts as a primer for the avian myeloblastosis virus DNA polymerase when Rous sarcoma virus heated 70S RNA is used as a template, offering evidence that it can hybridize to the RNA. The possible use of such an oligodeoxynucleotide hybridization competitor to inhibit Rous sarcoma virus replication is described in the preceding paper [Zamecnik, P. C. & Stephenson, M. L. (1978) Proc. Natl. Acad. Sci. USA. 75, 280--284].


Periodic loss of reactivity of a myeloma tumor with cytotoxic thymus-derived lymphocytes. During each transplantation passage of a line of mouse myeloma tumor MOPC-315 through syngeneic (BALB/c) hosts, the tumor cells lose reactivity with cytotoxic thymus-derived lymphocytes directed against products of the BALB/c major histocompatibility complex (H-2d) and regain reactivity on transfer to fresh hosts. In contrast to this cyclical change, the tumor cells remain uniformly reactive with anti-H-2d alloantisera throughout the transplantation cycle.


Subcellular fractionation of intra-axonally transport polypeptides in the rabbit visual system. We analyzed the subcellular distributions of proteins that are transported down the axons of rabbit retinal ganglion cells and compared these distributions to those of enzyme markers for endoplasmic reticulum, plasma membrane, and mitochondria. The proteins of each of five previously identified transport groups were uniquely distributed through the subcellular fractions, suggesting that each transport group is associated with different subcellular organelles. In particular, all of the observed group I polypeptides (the most rapidly moving, group, maximum velocity greater than 240 mm/day) were associated with material of hydrodynamic properties similar to those of the plasma membrane. The proteins of group II (maximum velocity = 34--68 mm/day) were heterogenous in their subcellular distributions but included mitochondrial proteins. Groups III and IV (maximum velocity = 4--8 and 2--4 mm/day, respectively) included materials that may be involved in motile processes; group V (maximum velocity = 0.7--1.1 mm/day) contained material of very high density which may be associated with neurofilaments.


Physicians' attitudes toward early intervention. Fourteen pediatricians serving Boston neighborhood health centers and fourteen pediatric neurologists in the greater Boston area were interviewed in order to determine their attitudes toward early intervention for infants with developmental disorders. The two groups differed significantly over the potential of early treatment for promoting optimal motor, cognitive, and affective development in the disadvantaged infant. Both groups viewed prevention of musculoskeletal deformities as appropriate for physically handicapped children and supported early intervention for the parents. When referral for treatment was delayed or withheld, the prime reasons given were an expression of a maturationist viewpoint, a desire for a definitive diagnosis prior to initiating treatment, the assumption that an adequate family can function without professional involvement, and misinterpretation of professional roles.


Effect of heptabarbital on bioavailability of bishydroxycoumarin in the rat. The influence of oral administration of heptabarbital on the gastrointestinal absorption of bishydroxycoumarin (BHC) was investigated in rats. Pretreatment of the animals with 20 mg/kg heptabarbital twice dialy for three days prior to the administration of 50 mg/kg BHC resulted in a 40% decrease from controls in the area under the BHC plasma concentration time curve (AUC). Preliminary studies indicated that a similar decrease in gastrointestinal absorption of BHC could be seen following a single oral dose of heptabarbital administered concomitantly with the anticoagulant.


Age-dependent presence of fetuin in bovine nasal secretion. At least two age-dependent proteins were detected in nasal secretion (NS) of young calves by crossed immunoelectrophoresis. One of them was identified as fetuin. The other one was not identified. The concentration of fetuin in NS was found to increase in cases where a sample of NS had been obtained at birth and a second sample taken within 11 days. Thereafter a decrease was always found towards day 15 or 30.