Protein patterns of the cerebrospinal fluid in children with cerebral palsy. Cerebrospinal fluid in 69 children with cerebral palsy (CP) of different etiology was examined by an improved method of agarose-gel electrophoresis. The protein pattern was normal in 12 cases only. In children with CP caused by congenital and postnatal infections raised psi-globulin fractions and the appearance of oligoclonal phi-globulin bands were the most obvious finding. In patients with CP due to malformations or perinatal brain damage and in children with CP of unknown etiology decreased prealbumin and increased albumin values were combined with hydrocephalus e vacuo in about half of the cases. Less frequent changes of the beta-globulin fractions were found, an increase of the beta-fraction being more rare than a decrease of the tau-globulin.


Neurological and developmental findings in children with optic disc drusen. In ophthalmoscopy, optic disc drusen often bear a fallacious resemblance to true papilloedema and some of those affected have neurological disorders. Adequate realization among neuropaediatricians of the relation between drusen and these disorders may save some children from neuroradiological and neurosurgical investigations, often considered unavoidable for exclusion of an intracranial process. Systematic evaluation in a series of 50 children with optic disc drusen showed that neurological disorders are common even in subjects brought to examination for other reasons than neurological symptoms. The findings showed accumulation into two subgroups. There was a group of 15 clumsy children with learning difficulties and delayed development of speech. Another group consisted of 20 children with sudden convulsions and/or headache and vomiting, and with EEG abnormalities but otherwise normal neurological findings. In addition, there were a subgroup with miscellaneous abnormalities and a minor group of children in whom no abnormalities were found except for the eye anomaly.


A survey of dental radiology evaluation in American dental schools. The variety of evaluation methods used in American dental schools illustrates that little uniformity exists. Only through a continuing program of evaluation can the strengths and weaknesses of any course be identified. Few generalizations can be made that are valid for all dental schools; this suggests that each school and its faculty members utilize methods of evaluation that are developed on a local level. The lack of uniformity in the evaluation process of the schools surveyed may also serve as an indicator for the lack of uniformaty of cirriculum. Development of clearly defined objectives would be beneficial in establishing reliable methods of evaluation. When such evaluations are analyzed, the need for changes in the curriculum becomes apparent. It should be noted that with every change,  whether it be in the students, the faculty, or the cirriculum, a new evaluation becomes necessary.


Absence of an alpha two-macroglobulin-protease complex in cystic fibrosis. The present study using immunologic methodology confirms previous observations from this laboratory of an absence of a protease component with arginine esterase activity in plasma of patients with cystic fibrosis. In this study, the pooled plasma from control individuals was activated and partially purified after adsorption on columns of soybean trypsin inhibitor conjugated to Sepharose 4B followed by elution with benzamidine. The fraction was further purified by isoelectrofocusing on polyacrylamide gels. Proteins around the pI range of 5.5 were eluted and utilized to prepare an antiserum. Immunoelectrophoresis of activated plasma samples from control subjects and patients with cystic fibrosis was performed utilizing the antiserum. In controls, four precipitin arcs with residual esterase activity were observed, whereas only three were seen in plasma from patients with cystic fibrosis. Double gel diffusion experiments using specific antisera ruled out the presence of trypsin, chymotrypsin, plasminogen, prothrombin, C1 esterase, alpha one-trypsin inhibitor, and inter-alpha-trypsin inhibitor in the concentrated benzamidine eluate. The antisera to alpha two-macroglobulin gave an immunoprecipitate which was readily stained for proteolytic activity. On immunoelectrophoresis, the alpha two-macroglobulin precipitin band corresponded to the band absent in plasma of patients with cystic fibrosis. In contrast, the alpha two-macroglobulin levels were similar in plasma of control subjects and patients with cystic fibrosis. Using the antiserum to the protein fractith proteolytic activity could be demonstrated in control plasma. One specific enzyme-active "rocket" was absent in plasma of patients with cystic fibrosis. In a double blind study of 15 control samples and 15 samples from patients with cystic fibrosis, a specific "rocket" was shown to be present in 13 control samples and absent in 14 cystic fibrosis samples. alpha two-Macroglobulin was determined by both an immunologic procedure and by its trypsin binding (trypsin protein esterase concentration). The ratio of the immunologic assay to the biologic activity assay was 90 for the normal plasma samples and only 65 for cystic fibrosis samples.


Social learning approaches to health education: utilization of infant auto restraint devices. There has been little critical evaluation of which methods achieve the desired results in health education. Using purchase of proper infant auto restraint devices as an objective, we assessed the effectiveness of three educational approaches. Only 37% of control families had purchased an acceptable car seat, compared to 54% of those who received descriptive literature alone, 71% of those that received literature and had been shown a descriptive film, and 60% of those who were given literature, shown the film, and been provided with a demonstration of the seat. Purchase of car seats was positively correlated with social class and knowledge of auto safety, but not with a history of auto accidents. Utilization of auto restraint devices is an example of how social learning theory can be applied in health education.


Structural and functional characteristics of individual phrenic motoneurons. Intracellular recording and staining techniques were applied to the study of cat phrenic motoneurons. Spontaneously driven phrenic cells possessed individualistic depolarization and spiking patterns that were a function of the conduction velocity in the different motor axons. Staining of phrenic motoneurons with Procion yellow indicated that fast conducting cells with small slow-wave depolarizations were large in size while slow conducting cells with large depolarizations were small in size. This implicated differences in membrane input resistance between large and small cells, although an unequal distribution of inputs to the individual components could not be discounted. On the average, phrenic motoneurons had a smaller dendritic surface area and smaller dendritic dominance than lumbosacral motoneurons. These factors help to explain the higher membrane resistances and longer time constants of phrenic cells. Phrenic dendrites were found to project in all directions away from the cell body and form ellipsoidal receptive fields that overlapped with other phrenic fields. It is speculated that the close approximation of phrenic dendrites with one another could, in part, be responsible for the high degree of synchronization among the different phrenic units.


Observations on template-specific conditions for DNA synthesis by avian myeloblastosis virus DNA polymerase. The effects of Mg++, Mn++, and KCl addition, individually and in combination, on the rate of DNA- and RNA-primed DNA synthesis by avian myeloblastosis virus DNA polymerase (reverse transcriptase) using a variety of natural and synthetic template-primer combinations were examined. Optimal divalent cation concentrations were found to vary by as much as 10-fold depending upon the template-primer used to direct synthesis. Addition of KCl to reaction mixtures containing optimal divalent cation concentrations produced stimulation or inhibition of DNA synthesis which was also template-specific. DNA synthesis on the modified template poly (2'-0-methylcytidylate) was uniquely stimulated by combinations of divalent cations. With Mg++ as divalent cation, deviations from classical Michaelis-Menten kinetics of substrate saturation were observed with all template-primers tested.


Role of purine N-3 in the biologic activities of poly(A) and poly(I). Poly(c3A) (poly 3-deazaadenylic acid) and poly(c3I) (poly 3-deazainosinic acid) differ in biological reactivity from their parent compounds poly(A) and poly(I) and from their 7-deaza counterparts poly(c7A) and poly(c7I). Three parameters of biological reactivity were evaluated : (1 degree) interferon induction, (2 degrees) anti-complement activity, (3 degrees) reverse transcriptase inhibition. Unlike poly(A)-poly(U), poly(I)-poly(C) and poly(I)-poly(br5C), the mixtures of poly(c3A) + POLY(U), poly(c3I) + poly(C), and poly(c3I) + poly(br5C) failed to elicit an interferon response in "super-induced" primary rabbit kidney cells; Poly(I) and its analogs poly(c3I) and poly(c7I) inhibited hemolytic complement activity, whereas poly(A) and its analogs poly(c3A) and poly(c7A) failed to do so. Both poly(I) and poly(c7I), but not poly(c3I), lost their anti-complement potency when annealed to either poly(C) or poly(A)-poly(U). Similarly, poly(I) and poly(c7I), but not poly(c3I), suppressed the interferon inducing ability of poly(A)-poly(U), suggesting that both poly(I) and poly(c7I), but not poly(c3I), added to poly(A)-poly(U) to form a triple-helical structure. Poly(I), poly(C7I) and poly(c7A)exerted a distinct inhibitory effect on turine leukemia virus, while under the same conditions poly(c3I) and poly(c3A) showed little, if any, inhibitory effect.


Poly 2'-O-ethylcytidylate, an inhibitor and poor template for AMV reverse transcriptase. Poly(2'-O-ethylcytidylate) is a poor template-primer for purified avian myeloblastosis virus reverse transcriptase; the relative activities of the template-primers poly(C)-oligo(dG), poly(Cm)-oligo(dG) and poly(Ce)-oligo(dG) are 23:16:1. A mixture of poly(Ce) and poly(dI) is inactive as template-primer, in agreement with the observed inability of these to form a helical complex. By contrast the inactivity of poly(Ce)-poly(I) is shown to be due to the influence of the 2'-O-ethyl residue. Poly(Ce) inhibits poly(A)-oligo(dT)-directed polymerase activity, with Ki = 3 muM, but marked inhibition with poly(A)-poly(dT) occurs only at low concentrations of the latter. Poly(Ce) did not inhibit template-primer activity of poly(C)-poly(dI) and poly(dC)-poly(dI). Qualitative physico-chemical studies show only partial complex formation between oligo(dG) and poly(C) and its 2'-O-alkyl analogues. This is discussed in relation to the widespread use of poly(C)-oligo(dG) as the template-primer for reverse transcriptase.


An evaluation of a procedure for the isolation of myelin basic protein (BP). A detailed description and stepwise evaluation of a procedure that can be used to obtain myelin basic protein (BP) from whole brain is presented. The procedure involved the 0.001 MHC1 extraction of whole brain pre-treated in a sequential manner with chloroform-methanol (2:1 v/v), acetone, and deionized water. This is followed by a precipitation of the extract at pH 9.0, and gel filtration of the supernatant in 0.01 M HC1. Yields of canine and porcine BP and their disc gel evaluations are presented at several key points in the procedure. The final products possessed a high degree of homogeneity when examined on SDS gels stained with commonly used protein stains. When compared with six SDS-gel marker-protein standards, the canine and porcine final products had mobilities that correspond to an apparent molecular weight of 18,5000 +/- 5%. Quantitative binding of 125I-labeled canine and porcine BPs with standardized rabbit anti-BP antisera gave comparable results. Immunoelectrophoretic and immunodiffusion examinations demonstrated single components and complete identity. The canine and porcine BP's also reacted fully with syngeneic anti-BP antisera raised in Lewis-strain rats. The canine BP was tested for encephalitogenicity in Lewis-strain rats and found to be comparable to rat BP in producing experimental allergic encephalomyelitis.


Platinum binds selectively to phosphorothioate groups in mono- and polynucleotides: a general method for heavy metal staining of specific nucleotides. Platinum binding to nucleoside phosphorothioates has been examined to determine their suitability as heavy metal labeling sites for the potential electron microscopic sequencing of nucleic acids. The complex platinum terpyridine nitrate forms a 1:1 adduct with either adenosine or uridine monophosphorothioate. Spectroscopic evidence strongly indicates the presence of platinum-sulfur bonds. Both platinum terpyridine nitrate and chloroterpyridineplatinum(II) bind to poly(sA-U), a polymer prepared from adenosine 5'-O-(1-thiotriphosphate) and UTP. Binding to the sulfur atoms of the phosphorothioate groups is quantitative, as shown by double label experiments using [35S]poly(sA-U) and [3H]chloroterpyridine-platinum(II). Similar experiments with [14C]poly(A-U) indicated no platinum binding. No evidence of nicking or loss of sulfur from poly(sA-U) could be detected after platinum binding. The phosphorothioate group is a strong, highly selective binding site for platinum in polynucleotides. Previous studies have demonstrated quantitative enzymatic incorporation of phosphorothioate groups into a polynucleotide adjacent to a specific base [Matzura, H. & Eckstein, F. (1968) Eur. J. Biochem. 3, 448-452]. The use of heavy metal-labeled phosphorothioate groups for the sequencing of nucleic acids by electron microscopy therefore appears feasible.


Small fragments from the A subunit of cholera toxin capable of activating adenylate cyclase. Exposure of cholera toxin to membrane particles prepared from sarcoma 180 cells gives rise to a variety of fragments which are capable of activating adenylate cyclase [ATP:pyrophosphate-lyase (cyclizing), EC 4.6.1.1]. A major component of these fragments has an apparent molecular weight in the 8,000-10,000 range. The smallest stimulatory fragment has a molecular weight of approximately 1400. The small size of the fragments is confirmed by Sephadex gel filtration, in the presence of either sodium dodecyl sulfate or formic acid. These fragments are produced from holotoxin or its A subunit by protease(s) found in sarcoma membrane particles. Production of fragments appears optimal in 40-60 min at 30 degrees and pH 7, and is prevented by protease inhibitors. The ability of the small fragments to activate adenylate cyclase is reversed by anti-holotoxin, but not anticholeragenoid, antibodies. These fragments require NAD for the activation of adenylate cyclase and are fully active after heating at 90 degrees for 5 min (pH 7).


Demonstration of the inhibitory effect of human alpha-fetoprotein on in vitro transformation of human lymphocytes. We have studied the effects of human alpha-fetoprotein (HAFP), isolated from the serum and ascitic fluid of a hepatoma-bearing patient, on the in vitro transformation of human peripheral blood lymphocytes by a variety of mitogenic stimuli. At a concentration of 2.5 mg/ml, HAFP inhibited the lymphocyte response to phytohemagglutinin, concanavalin A, and rabbit anti-human thymocyte serum, but failed to inhibit the response to pokeweed mitogen. HAFP was able to inhibit the one-way mixed lymphocyte culture at concentrations of 250-500 mug/ml, but failed to inhibit at 100 mug/ml. Exposure of lymphocytes to 2.2 mg/ml of HAFP for 18 hr did not result in significant lymphocytotoxicity, and such cells washed free of HAFP were fully capable of participating in the mixed lymphocyte culture. HAFP did not inhibit lymphocyte E-rosette formation. Fetal HAFP was more effective in inhibiting human lymphocyte responses than hepatoma HAFP. These experiments support the suggestion that HAFP plays an important immunoregulatory role during fetal development, possibly through the suppression of thymus-derived lymphocyte responses to antigenic stimuli; they also suggest that there are important differences in the biological properties of hepatoma and fetal HAFP.


Determinants of antigenic molecules responsible for genetically controlled regulation of immune responses. The ability of mice bearing the H-2S haplotype to develop helper responses to the random copolymer of Glu,Ala while they developed suppressor responses to the terpolymer of Glu,Ala,Tyr suggested the crucial role of tyrosine in these peptides. On the basis of various considerations, it was postulated that many of the tyrosine residues in Glu,Ala,Tyr would be localized at the NH2-terminal end of the molecule. To verify this hypothesis, a block terpolymer composed of a short sequence of homopolymer tyrosine covalently bound to the random copolymer of Glu,Ala was synthesized. The present studies, using this block terpolymer, demonstrated that the chemical determinants stimulating helper and suppressor responses are distinct and can be present simultaneously in the same molecule. Thus, addition of COOH-terminal tyrosine residues to the Glu,Ala polypeptide converted this immunogenic molecule to an immunosuppressive molecule in mice bearing the H-2S haplotype. The mechanism by which these short sequences of tyrosine influence H-2-linked immune responses remains to be determined.


Developmental screening clinics are a luxury. A survey of children under 5 in an urban practice has been described and the means of detection of developmental abnormalities identified. It was found that 81% were queried by nonclinic means and only one child out of 150 had a minor abnormality which might otherwise have gone undetected if not seen at a screening clinic. These developmental screening clinics are expensive to run and yield very little return.


Evidence for a gonadotropin from nonpregnant subjects that has physical, immunological, and biological similarities to human chorionic gonadotropin. Substances from urinary extracts of normal, nonpregnant subjects and human pituitary gonadotropin preparations were found to react similarly to human chorionic gonadotropin (hCG) in a radioimmunoassay system that is highly specific for hCG and without crossreactivity to human luteinizing hormone (hLH). The antiserum was produced in a rabbit immunized with a bovine albumin conjugate of the unique carboxyl-terminal peptide (residues 123-145) isolated from a tryptic digest of the reduced, S-carboxymethylated hCGbeta subunit. The antibody recognition site on the peptide was found to reside on the last 15 amino acid residues of the carboxyl-terminal peptide, as evidenced by the competitive binding activities against 125I-labeled hCG of a series of peptides chemically synthesized according to the carboxyl-terminal sequence of HCGbeta. In order to elucidate the nature of the crossreacting substance in urinary extracts, a human postmenopausal urinary preparation (Pergonal) and a kaolin-acetone extract of urine from a patient with Klinefelter's syndrome were subjected to gel chromatography on Sephadex G-100. The results indicate that fractions showing immunocrossreactivity with the antiserum to hCGbeta-carboxyl-terminal peptide coeluted with 125I-labeled hCG which was separated distinctly from hLH. The same fractions from this postmenopausal urinary gonadotropin preparation exhibited in vitro biological activity proportional to the immunocrossreactivity of the hCG-specific antiserum. Concentration of postmenopausal women's urine by acetone precipitation retained approximately five times more immunoreactivity per unit volume than kaolin-acetone extraction, when assayed with the antiserum to hCGbeta-carboxyl-terminal peptide.


Developmental assessment is a necessity. In this paper I shall discuss a typical developmental disorder, showing how its manifestations vary throughout childhood, and comment upon the basic criteria for an assessment service.


The binding of the optical isomers of methadone, alpha-methadol, alpha-acetylmethadol and their N-demethylated derivatives to the opiate receptors of rat brain. The optical isomers of methadone, alpha-methadol, alpha-acetylmethadol and their N-demethylated derivatives have been systematically studied for their effects on the binding of 3H-dihydromorphine (3H-DHM) and 3H-naloxone (3H-NLX) to opiate receptors in rat brain homogenate. The relative affinities of these agents in competing for both 3H-DHM and 3H-NLX binding parallel their analgesic effects. 1-Methadone is about 30 times as effective as d-methadone in competing for both 3H-DHM and 3H-NLX binding sites. The reduction of 1-methadone to alpha-d-methadol and subsequent N-demethylation to alpha-d-normethadol reduce its effectiveness as indicated by the increase in the IC50 values for both 3H-DHM and 3H-NLX binding. The reduction of d-methadone followed by N-demethylation produces a potent derivative, alpha-1-normethadol, which has IC50 values on 3H-DHM and 3H-NLX binding similar to those of 1-methadone. The affinity of alpha-1-acetyl-methadol on the binding of both 3H-ligands falls between those of 1-methadone and d-methadone, and increases as it is N-demethylated. alpha-d-Acetyl-methadol is more effective than alpha-1-acetylmethadol in competing for both 3H-ligands from the opiate receptors, and its affinity, unlike that of alpha-1-acetylmethadol, decreases when it is N-demethylated. The affinities of the methadone isomers and related compounds on the binding of 3H-NLX fall in the presence of Na+. The latter property indicates the agonistic nature of this series of druges.


Morphologic and antigenic maturation of lymphocytes in the mouse thymus in vitro. A simple technique for organ culture of the thymus of 14-day-old mouse embryos is decribed. It allowed a characteristic time-dependent development of the immature thymus into a lymphoid thymus with large numbers of nondividing small lymphocytes. Most of these cells carried the T-lymphocyte antigenic markers Thy-1 and TL, and all expressed H-2 antigens as determined in cytotoxicity assays. They probably developed from precursor cells without detectable Thy-1 and TL. This development appeared to be dependent on the thymic microenvironment, since it also occurred in serum-free organ cultures and in cultures with medium supplemented with serum from nude mice.


Prenatal detection of anencephaly.

Prenatal detection of anencephaly. Fetal anomalies were suspected clinically in 3 patients, during the second trimester of pregnancy, due to advanced maternal age, hydramnios and abnormal vaginal bleeding respectively. Sonography revealed anencephaly in all of them, confirmed by roentgenograms and also by high amniotic fluid leveal alpha-fetoprotein in 1 of them. The antenatal diagnosis was confirmed in the 3 abortuses. Early screening of all pregnancies at risk for neural tube malformations is strongly recommended.


Detection of beta2-microglobulin in sera from diverse vertebrate species by the use of a heterologous radioimmunoassay. The strong cross-reaction between rabbit and human beta2-microglobulins, as detected by goat antisera, has permitted the development of heterologous radioimmunoassays to detect homologues of beta2-microglobulin (beta2mu) in the sera of various vertebrate species. The assay based on the inhibition of the binding reaction between goat anti-rabbit beta2 mu and human beta2 mu divided the sera examined into two groups, one giving high levels of inhibition (approximately 80%). and the other lower levels (approximately 20%). There was no evident correlation between this division and phylogenetic classification. When the same group of sera was tested using the assay based on the inhibition of binding of rabbit beta2 mu to goat anti-human beta2 mu, a wide range of maximal levels of inhibition was observed, with clustering of phylogenetically related species.


Prozone effects in indirect immunofluorescence. A marked prozone effect was observed in indirect immunofluorescence (IFL) with rabbit antisera against rat renal proximal tubular epithelial brush border (BB) antigens: anti-BB antibodies were not detectable if used in high concentrations but were readily demonstrated if sufficiently diluted. The prozone effect occurred in spite of demonstrated binding of anti-BB antibodies to target antigens. No prozone was seen when direct IFL was used, and 'cross-reacting' antigens in small-intestinal epithelial brush border were detectable by indirect IFL without prozone. It was concluded that the anti-immunoglobulin conjugates were unable to reach antigenic determinants on tightly clustered immunoglobulin molecules. Thus prozone effects seem to depend primarily on the density of antigen determinants in the target tissues.


Shared idiotypic determinants on B and T lymphocytes reactive against the same antigenic determinants. V. Biochemical and serological characteristics of naturally occurring, soluble antigen-binding T-lymphocyte-derived molecules. Normal rat lymphocyte populations house a high percentage of lymphocytes with idiotypic, antigen-binding receptors for the major histocompatibility complex antigens of the rat. These receptors can be isolated from normal serum or lymphocyte supernatants. Idiotypic, antigen-binding molecules released from normal Lewis lymphocytes were thus isolated using anti-(Lewis anti-DA) immuno adsorbents. Analysis by SDS polyacrylamide electrophoresis using molecules labeled by external or internal means (125I or 3H) demonstrated that B lymphocytes produce molecules of 'conventional' 7S-8S IgM type. T-lymphocyte-derived molecules had a molecular weight of around 150,000 and consisted of two chains of similar size. Such single chains would succumb to proteolysis by normal serum factors to yield fragments in the size range of 30,000-40,000 daltons. All three groups of T-cell-derived molecules expressed both antigen-binding and idiotypic markers. No evidence was obtained that any light chains are linked to the T-receptor molecules. Serological analysis of the T-cell molecules failed to prove the existence of any determinants of constant immunoglobulin type, nor did these molecules express antigenic markers of major histocompatibility complex types.


Intrahepatic localization of the surface (HBsAg) and core (HBcAg) antigenic determinants associated with hepatitis B virus in biopsy samples from patients with liver disease. 109 biopsy samples from 35 HBAg serologically positive and 74 negative patients were examined by IFL for the presence of the surface and core antigenic determinants associated with the Dane particle. In no serologically negative case was specific IFL detected. Different patterns were observed in serologically positive patients: negative in acute hepatitis, strongly positive cytoplasmic HBs fluorescence in chronic HBAg carriers with normal liver, and discrete HBsAg parenchymal and mesenchymal staining and variable HBcAg staining in chronic liver disease, with HBsAg appearing more frequently in active and HBcAg in active disease. These results are compared with recent reports in this field and the clinical significance of the intrahepatic localization of HBAg is discussed.


Placental localization of human pregnancy--associated plasma proteins. Frozen sections of human placenta were examined for the presence of four human pregnancy proteins, pregnancy-associated plasma proteins A and C (PAPP-A and PAPP-C), human chorionic somatomammotropin (hCS), and pregnancy zone protein (PZP), by the indirect immunofluorescence technique. Monospecific rabbit antiserums to PAPP-A, PAPP-C and hCS all stained the trophoblast cytoplasm equivalently in a continuous layer, usggesting that the same trophoblast cells synthesize all three pregnancy proteins. In contrast, PZP was localized in blood vessel walls, parenchymal structures within the villous, as well as in the trophoblast cytoplasm. Its distribution in the latter was relatively inhomogeneous, tending to be more intense on the basement membrane side.


Studies on the diagnosis and treatment of human filariasis in Rhodesia. Experiences in Rhodesia with various recovery techniques available for the laboratory diagnosis of infections with Dipetalonema perstans and Wuchereria bancrofti are discussed. A diagnostic laboratory regimen for routine filarial investigations is suggested. Included are preliminary observations on the use of mebendazole (Vermox) for the treatment of D. perstans infections.


Prolongation of allograft and xenograft survival by pretreatment with prednisolone and donor-specific antigen. Sprague-Dawley rats were grafted with skin of Wistar rats. Surprisingly, pretreatment with donor-specific spleen homogenate and semisoluble antigens combined with prednisolone resulted in a prolonged survival of the skin grafts. The same effects were observed after transplantation of rabbit skin to Sprague-Dawley rats. Lewis rats were grafted with kidneys of LewisX brown-Norway F1 rats and simultaneously subjected to bilateral nephrectomy. The recipients were pretreated with soluble and with semisoluble donor-specific antigen. The recipients survived in definitely after pretreatment with semisoluble antigen combined with prednisolone. Combined administration of soluble antigen and prednisolone was ineffective. In the canine kidney allografts, the recipients were pretreated with semisoluble donor-specific spleen antigen; in the canine liver allografts, they were given soluble and semisoluble antigen. The canine kidney recipients survived significantly longer when they had been pretreated with antigens combined with high doses of prednisolone. Pretreatment with high doses of prednisolone alone did not prolong the survival of kidney grafted rats and dogs, and pretreatment with antigen alone had no effect on the survival of kidney grafted dogs. Pretreatment with antigen alone had no effect on the survival time of liver allografts in dogs, but the recipients of the liver grafts survived longer than and controls--some of them for more than 1,200 days--when they had been pretreated with antigens combined with prednisolone. In this respect, the semisoluble antigen was more effective than the soluble form. After three spaced antigen injections, the lymphocytotoxic antibody titers on the day of the grafting varied, but they were always low. There was no correlation between the titers on the day of transplantation and the duration of survival. Recipients pretreated with antigen showed lower postoperative antibody titers than those without antigen.


Specificity of antisera to human fibrinopeptide A used in clinical fibrinopeptide A assays. Distinction between fibrinopeptide A (FPA) and larger polypeptides containing the FPA sequence is critical for the interpretation of clinical results with FPA immunoassay methods. Therefore, the immunochemical reactivity of 14 rabbit anti-FPA sera with six different FPA containing antigens was studied in detail. Antigens tested included: fibrinogen; fragment E of fibrinogen; the amino-terminal disulfide knot of fibrinogen; Aalpha 1(Ala)-51(Met); Aalpha 1(Ala)-23(Arg); and, FPA. Synthetic partial sequences of FPA were also tested. The 14 FPA-specific antisera were divided into 3 distinct categories with: I, FPA immunoreactivity of larger polypeptides containing FPA approximately 1/100 of FPA on a molar basis, II, FPA immunoreactivity of the larger polypeptides intermediate between I and III; and III, FPA immunoreactivity of the larger polypeptides approximately equal to that of FPA on a molar basis. The antigenic determinants of a category I antiserum (R 2) are included in Aalpha 7(Asp)-16(Arg) with Asp(7), Phe(8) and Arg(16) being essential. When attached to FPA, the sequence Gly(17)-Arg(23) decreases the immunoreactivity of FPA with category I antisera 100-fold. The practical consequence of these findings is that, when category III antisera are employed, both FPA and larger FPA-containing polypeptides are equally immunoreactive. Since thrombin treatment of the larger polypeptides does not alter their immunoreactivity, category III antisera cannot discriminate between FPA and the larger polypeptides. On the other hand, with category I antisera, although the immunoreactivity of FPA itself is unaltered by thrombin treatment, larger polypeptides [e.g., Aalpha 1(Ala)-23tArg)] show a 100-fold increase in immunoreactivity following thrombin treatment and thus can readily be identified and separately quantitated. It is concluded that antisera with the specificity of category I are essential for the specific and accurate measurement of FPA, and for its distinction from larger FPA-containing polypeptides, in clinical plasma samples.


Evidence for an ester bond between thrombin and heparin cofactor. Heparin cofactor, a thrombin inhibitor, is purified from human plasma by affinity chromatography on heparin-agarose. The nature of the binding between thrombin and the inhibitor is studied by treatment of the complex with 6 M guanidinium chloride, hydroxylamine, and dilute alkali. The complex is not dissociated during gel chromatography in 6 M guanidinium chloride. This result supports an earlier proposal that formation of the complex includes the formation of a covalent bond. Treatment of dodecyl sulfate-denatured complex with hydroxylamine results in dissociation of the complex to yield free thrombin and heparin cofactor. Hydroxylamine does not dissociate the complex unless it is denatured. The complex is also dissociated in dilute sodium hydroxide (pH 12) solutions. These results indicate that the covalent bond between thrombin and the inhibitor is a carboxylic ester.


HLA-D antigen of Japanese origin (LD-Wa) and its association with Vogt-Koyanagi-Harada syndrome. One homozygous cell "Wa" for HLA-D alleles, associated with HLA-Bw22J (Japanese specific variant of HLA-Bw22), was found in a Japanese family. The HLA-D antigenic specificity defined by this cell has been tentatively named LD-Wa. Phenotype frequency of LD-Wa in the Japanese population was 0.16 (13/81). HLA-D typing for LD-Wa was performed in patients with the Vogt-Koyanagi-Harada syndrome (Harada's disease) with which Bw22J is strongly associated (15/35, 42.9%; corrected P less than 0.02). LD-Wa was presented in 22 of 33 patients (33.7%; P less than 0.0000003). The association of LD-Wa with Harada's disease was stronger than that of Bw22J, and this implies that there might be one of the disease susceptibility gene loci affecting the development of this disease more closely linked to an HLA-D gene locus than the HLA-B locus.


HLA and juvenile chronic polyarthritis.

HLA and juvenile chronic polyarthritis. HLA antigen have been identified in patients with juvenile chronic polyarthritis (J.C.P.) (n = 35). In J.C.P. the incidence of antigen B27 (57%) was found to be higher than in a normal population (n = 1,000). On recent evaluation of the clinical status, serology and x-rays, the patients with juvenile chronic polyarthritis who has been followed up many years, could be subdivided into four groups: Group 1: J.C.P. evolving to ankylosing spondylitis (n = 3); Group 2: J.C.P. with sacroiliitis (n = 17); Group 3: J.C.P. without sacroiliitis (n = 9); Group 4: juvenile rheumatoid arthritis characterised by a positive serology (n = 6). Groups 1 and 2 were characterized by a high incidence of antigen B27 (19/20). Only one subject of groups 3 and 4 had antigen B27. Sex distribution in groups 1 and 2 was found to be similar to that found in ankylosing spondylitis in adults and sex distribution in groups 3 and 4 was found to be similar to that found in rheumatoid arthritis in adults. It is concluded that if B27 positive develop juvenile chronic polyarthritis they have a high risk of developing sacroiliitis and acute uveitis. The authors propose to give the name Still's disease to the total group of children presenting initial symptoms corresponding to the criteria of Ansell & Bywaters (1959). Follow-up of the cases permitted their further classification as juvenile ankylosing spondylitis, as juvenile chronic polyarthritis with or without sacroiliitis or as juvenile rheumatoid arthritis.


The major histocompatibility complex of rhesus monkeys. VI. Serology and genetics of Ia-like antigens. The serology and genetics of11 new cell surface alloantigens of rhesus monkeys are described: They are controlled by the mamor histocompatibility complex but are distinct from the conventional serologically defined (SD) antigens ofRhL-A. The new specifications are termed "Ia-like" because ofserological, immunocytological and other characteristics reminiscent of Ia-antigens of the mouse. Population and family analyses led to the postulation of two segregant series controlling eight of the 11 Ia-like specificities of the monkey. Strong linkage disequilibria with SD2 antigens and genetic mapping on the basis of segregation studies in recombinant offspring in the monkey families, places at least one of the two loci in the vicinity of the SD2 locus of RhL-A, not in the region of the major MLC or LD1 locus. For this and other reasons, the new B-cell alloantigens of rhesus monkeys are not believed to be similar to or associated with the stimulator antigens of LD1. The biological function(s) of the Ia-like antigens of primates are as yet unknown.


Specific MLC stimulation by cultured B cells. Pairs of cultured T and B lymphoid cells, derived from the same individual, were used as stimulating cells in MLC experiments. Blastogenic response to cultured B cells were much greater than those to the corresponding T cell line. These cells will provide a useful reagent for isolation and characterization of B cell specific antigens.


Antigenicity of methylated deoxyribonucleic acid. Methylated salmon sperm deoxyrinbonucleic acid (Me-ssDNA) prepared by using dimethyl sulfate was used to elicit antibodies in rabbits. The antisera obtained reacted specifically with Me-ssDNA but not with thermally denatured Me-DNA, native DNA, thermally denatured DNA or ribonucleic acids (RNAs). Antibody activity was demonstrable in both IgG and IgM fractions which were obtained by Sephadex G-200 column chromatography. A single precipitating line was formed by either of IgG and IgM fractions in the double diffusion test with Me-ssDNA.


Iron gallein elastic method---a substitute for Verhoeff's elastic tissue stain. A method for staining elastic fibers in formalin fixed, paraffin embedded sections is described. After deparaffinizing and dehydration, sections are stained for 30 minutes in a solution prepared by mixing equal parts of 1% gallein dissolved in ethylene glycol and absolute alcohol (1:4), and 1.16% aqueous ferric chloride in 1% hydrochloric acid. The sections are washed in water and then differentiated in 2% ferric chloride for 2 minutes. After washing in water, the sections are counterstained with a variant of Van Gieson's picric acid-acid fuchsin for 1 minute. The results are similar to Verhoeff's elastic stain with elastic fibers staining black. An advantage to this staining procedure is that visually controlled differentiation is not necessary.


Staining of keratin and keratohyalin with the reactive dye levafix red violet E-2BL. Demonstration of keratin in Zenker-fixed skin and in tissues stored in formalin can be difficult because such material is unsuitable for histochemical studies. A reactive dye, Levafix red violet E-2BL, proved useful for demonstration of keratohyalin and some types of keratin. Formalin-, Zenker- and methacarn-fixed sections were pretreated with alkaline alcohol, stained one hour at 60 C in an aqueous solution containing 0.25% Levafix red violet E-2BL plus 0.25% NaCl, rinsed in buffer solution pH 9, dehydrated and mounted. Keratohyalin granules and stratum corneum were colored red violet; hair and tonofibrils remained unstained. In sections prestained with Mayer's acid hemalum, keratohyalin was dark blue. Sulfonated monoazo dyes without reactive groups colored no tissue structures under the conditions of this technic; apparently, Levafix red violet E-2BL is bound via its reactive group. Polarization microscopic studies suggest binding of Levafix red violet E-2BL by an amorphous matrix of keratin. Correlations with chemical data indicate that the staining patterns parallel the distribution of proteins formed in the stratum granulosum.


A consistent phosphotungstic acid hematoxylin stain for glial fibers. After deceration, celloidinization and hydration, oxidize 10 micron paraffin sections for 15 min in a solution containing 0.3 g KMnO4 and 0.1 ml conc. H2SO4 per 100 ml distilled water. Wash in water and reduce in 5% oxalic acid until the sections are colorless. Wash thoroughly in water and place in 4% iron alum solution for two hours. Wash briefly in water and stain for two hours in phosphotungstic acid hematoxylin. Rinse briefly in 95% ethanol and dehydrate in n-butyl alcohol or absolute ethanol for 4 min with two changes, clear and mount. Glial fibers, myofibrils, red blood cells, etc. are stained blue while astrocyte cell bodies, collagen, etc. are stained red. This stain has proven highly consistent in a wide variety of astrocytic derangements. Despite the intensity of this PTAH modification, false positive staining was not observed.


A new method for easy demonstration of argyrophil cells. A new method for silver impregnation of endocrine cells of the gastrointestinal mucosa is described. It offers great reliability, eveness of impregnation, and, since it can be used on batches of slides, is also suitable for histology class and investigation material. The procedure for paraffin sections of formalin-fixed material is as follows: dewax and transfer to distilled water, leave in 0.5% silver nitrate solution for 2 hours at 60 C. Rinse in distilled water, then treat in Bodian developer (hydroquinone, 1 g; sodium sulphite, 5 g; distilled water, 100 ml) previously heated to 60 C. Rinse in running tap water, distilled water, and then re-impregnate for 10 minutes at 60 C in the same silver solution and reduce in Bodian's solution. Since the background is not impregnated by this method, sections may be counterstained by any basic anilin dye to bring out nuclei. A 0.1% kernechtrot solution was found very satisfactory in this respect. The granulations of argyrophil cells stand out sharply black against a red background.


Differential staining of tannin in sections of epoxy-embedded plant cells. A staining procedure is described for the light microscopic localization of ergastic tannins in epoxy sections of plant cells embedded for study by transmission electron microscopy. Callus and cell suspensions of Pseudotsuga menziesii and Pinus taeda fixed in glutaraldehyde:acrolein and then OsO4, followed by epoxy embedding, were sectioned 0.5 mum thick, stained on a glass slide with ethanolic Sudan black B at 60 C as described by Bronner, and then mounted in Karo syrup. Tannin deposits stained brownish-orange and were easily distinguished from lipid bodies of similar size, which stained dark blue to black, and from starch grains, which were unstained. The significance of this differential polychromasia was confirmed by transmission electron microscopy. This staining procedure should prove valuable in the cytoplasmic evaluation of the plant cell ergastics (especially tannins) via light microscopy whether or not electroc microscopic examination is intended.


[The use of nuclear medicine methods for the control of regional cytostatic therapy of neoplasms in the orofacial region]. Complementary to the well-known clinical, radiographic, histologic and biochemical methods for the control of the regional cytostatic chemotherapy of malignant tumours in the orofacial region, the authors describe by help of a case report the possible uses of nuclear-medical methods of examination. The following nuclear-medical methods of examination were employed: 1. bone scintigraphy, 2. perfusion scintigraphy, 3. tumour scintigraphy. The advantages of nuclear-medical methods consist in the relatively low load to which the patient is subjected and in the possibility of assessing qualitatively and quantitatively (in combination with current control methods) the distribution of the perfused chemotherapeutic agent and of evaluating the therapeutical success.


Antigenic relationships among Nigerian strains of West Nile virus by complement fixation and agar gel precipitation techniques. Using two serological techniques, eight Nigerian West Nile virus isolates were investigated to determine antigenic relationships among them, and to find out if these virus isolates were related to West Nile virus strains from the different zoogeographic areas of the world. One virus differed significantly from the seven other strains and was later found to be a strain of Usutu virus. The remaining strains were differentiated into two serological intratypic groups depending on their cross reactions with two strains which served as prototypes for each group. Five virus isolates which constitute one of the antigenic groups were found to be related to the Egypt 101 strain of West Nile virus originating from general Palearctic zone (European and Middle East). The other two virus isolates did not show any relationship to the strains from any of the different zoogeographic zones.


Blood coagulation and plasma fibrinolytic enzyme system pathophysiology in stroke. Plasma fibrinogen chromatography is a method for quantification of high molecular weight fibrinogen complexes (HMWFC), native fibrinogen and other fibrinogen derivatives in plasma. Enchanced formation of fibrin, intravascular coagulation, thrombus formation, etc., are reflected by elevation of plasma HMWFC, and the method distinguishes between subjects with normal and pathological rates of fibrin formation. Serial standard blood coagulation assays, including plasma fibrinogen chromatography, and neurological studies were performed on 220 patients admitted to a stroke unit. Findings from patients with cerebral infarction were compared against those of three control groups: (1)normals, (2)a stroke control group and(3)a stroke risk factor group. Plasma HMWFC findings were significantly (p less than 0.001) higher in the stroke risk factor group than in the normals. Plasma HMWFC values were significantly higher (p less than 0.001) in the cerebral infarction patients than in any of the control groups, and plasma fibrinogen, plasminogen, alpha1-antitrypsin and alpha2-macroglobulin also were significiantly higher (p less than 0.001) in the patients. The greater the degree of initial neurological deficit, the greater were plasma HMWFC values (p less than 0.001), and high HMWFC values were associated with poor clinical outcome. Plasma HMWFC values were significantly higher (p less than 0.001) in patients with intracerebral hemorrhage, subarachnoid hemorrhage and cerebral embolism. These findings docunment the fact that a high proportion of stroke patients have coagulopathy, characterized by pathological enhancement of fibrin formation.


Long-term anticoagulant therapy for TIAs and minor strokes with minimum residuum. One hundred seventy-eight patients with transient ischemic attacks (TIAs) or small strokes with slight symptoms persisting for more than 24 hours (incomplete recovery = IR) (TIA-IR) from both the carotid and the vertebrobasilar systems were treated with anticoagulants. Ten patients stopped the treatment because of severe side effects. Only one patient had a lethal cerebral infarction when the thrombotest values were above the therapeutic level; no other infarction happened during the treatment period. Moreover, the frequency of TIA decreased during the treatment, compared with descriptions of the natural course of TIA. One hundred four patients were observed for a mean of 21 months after the anticoagulant treatment ended. During the observation period, six patients had cerebral infarctions. This was a sixfold increase compared with the stroke incidence during treatment, and was almost identical with the incidence of strokes seen during the natural course of TIA. All the cerebral infarctions were in patients who had their initial TIA/TIA-IR from the carotid territory (within the same carotid artery which earlier had given symptoms). The investigation shows that long-term anticoagulant treatment is useful, especially in patients with carotid TIA/TIA-IR, and that this treatment should continue as long as the patients can manage it. In patients with vertebrobasilar symptoms of malignant character, it seems feasible to terminate the treatment after about one year. The mechanism of the anticoagulant treatment is obscure, but it does not appear to influence the progress of the atherosclerotic process.


[Serum acid phosphatase levels following prostatic massage (author's transl)]. We find an elevation of serum acid phosphatase levels 5 min after prostatic massage in only 10% of patients with prostatic adenoma. This increase is caused by stored prostatic secretion being pressed into the blood vessels. 60 min later these serum levels decrease. With some other patients a slower increase of phosphatase levels occurs; this increase, however, lasts for hours. This kind of increase is caused by prostatic fluid being forced into the interstitium where it is slowly absorbed. A combination of both kinds leads to a curve with two peaks; this could be demonstrated in two cases. No difference was seen in reaction of total acid phosphatase and prostatic phosphatase levels. The increase of phosphatase levels following prostatic massage was no sign of prostatic carcinoma.


Scanning electron microscopy of hyperplastic and neoplastic human prostate. Acinar cells of neoplastic prostate tissues are more heterogeneous in size and shape than benign hyperplastic cells when observed by scanning electron microscopy. Three types of acinar cells are recognizable by surface structure, cells with microvilli, cells without microvilli, and cells with membrane ruffles. The pitted cells previously seen in BPH tissues are probably artifactual. The identity of the crater cells is sitll in question.


Inhibition by estramustine phosphate on estradiol and androgen binding in benign and malignant prostate in humans. Specific cytoplasmic androgen and estrogen binding has been measured in human benign prostatic hypertrophy and carcinomatous tissue. In vitro support for the binding of estramustine phosphate (Estracyt) to both estradiol- and dihydrotestosterone-binding sites is presented, which in part could explain the clinical effect of estramustine phosphate when pure estrogenic compunds are not effective.


[Effect of temperature and proteolytic enzymes on the immunological properties of 2 acid neurospecific antigens]. The immunological properties of water-soluble acid neurospecific antigens A and D of the bull brain were studied as affected by heating and proteolytical enzymes (trypsin, mixture of trypsin and alpha-chymotrypsin, protease from Streptomyces griseus). The immunological activity of antigen A is not changed essentially after its heating for 20 min at a temperature of 70-80 and 24-hour proteolysis with trypsin. Heating at 90-100 C as well as hydrolysis with protease from Streptomyces griseus and mixture of trypsin with alpha-chymotrypsin for the same time decrease the immunological activity of antigen A. The immunological activity of antigen A lowers after heating for 20 minutes at a temperature of 60 C and the action of all the studied proteolytical enzymes for 24 hours. The data obtained evidence for protein nature of antigens A and D.


[Antigenic analysis and immunological studies of the uterotropic bacterial strains SH6 and 01 isolated from aborted cows]. The uterotropic strains SH6 and O1 were studied immunoelectrophoretically to establish their antigenic relationship. The more readily diffusing component of the O-antigenic fraction of the strains proved immunologically identical with LPA  of the R Brucella organisms, while the slower component showed common determinants with the O-antigenic fraction of Yersinia pseudotuberculosis. Immunodiffusion revealed that the O-antigenic fraction of the O1 strain and LPA of the S Brucella organisms contained common antigenic determinants having a weak immunologic activity, and this could explain the agglutination reaction after Wright and Huddleson of the sera from cows with infections caused by the uterotropic strains described. It was established through electrophoresis that in contrast to Yersinia enterocolitica 373 the O-antigenic fraction of the uterotropic strains is immunologically identical with LPA of the R Brucellae. An analogy is shown to exist between the immunologic activity of cow sera from infected animals and the A substance as well as at higher agglutination titers after wright of substance B as described by Amarasinghe.


[Potential sources of errors in the gas chromatographic determination of organochlorine pesticides]. It has been found that the reliability of the results obtained in the determination of chlororganic pesticides depends on the frequence of using reagents and adjuncts. When these means are not sufficiently pure peaks suggesting impurities appear in the blenk, which, during the retention time overlap those of some of the chlororganic pesticides (alfa-BHC, gamma-BHC, aldrin) and could mislead the analyst. The present work deals with the methods of purifying the reagents and adjunct material in order to obtain blenk containing no impurities.


[The effect of decomplementation on model experimental inflammatory responses (rat paw oedema) (author's transl)]. An investigation was carried out of the effect of decomplementation by either aggregated human gamma globulin (aggHGG) -- activating the classical complement cascade -- or cobra-venom factor (CVF) -- acting through the alternative pathway -- on rat paw oedema induced by various agents. The direct, passive Arthus reaction (DPA) and dextran-and carrageenin-induced oedema were inhibited by both decomplementing measures, whilst serotonin-and formalin-induced oedema remained unaffected. The paw swelling induced by synthetic bradykinin was significantly reduced only by aggHGG, not by CVF. These observations strongly support -- but do not yet prove -- the assumption that complement is involved in the development not only of the DPA but also of the responses to the polysaccharides, dextran and carrageenin in the rat. The partial inhibition of bradykinin-induced paw oedema by aggHGG and not by CVF might be explained by a densensitization or blocking of bradykinin receptors by a peptide derived from the early-reacting complement components (C 1, 4, 2). Investigations of the effect of aggHGG and CVF on blood pressure, haematocrit, partial thromboplastin time, plasmakininogen and kinase activity, leucocytes and thrombocytes indicate that non-complement-dependent factors are unlikely to be involved in the observed inhibition of experimental rat paw oedema.


Passive immunization against exposure to hepatitis B virus in the military: potential and possibilities. The value of standard γ-globulin with a low titer of antibody to hepatitis B surface antigen (anti-HB(s)) vs hepatitis B immune globulin (HBIG) in prevention of icteric hepatitis B in the military is unclear. Although recent studies have shown a decrease in icteric hepatitis after administration of both types of γ-globulin in populations where acquisition of hepatitis B virus (HBV) is most likely the result of nonparenteral transmission, the data pertaining to parenteral exposure suggest that HBIG delays the incubation period of HBV and decreases the development of passive-active immunity. Since no studies have demonstrated efficacy of standard γ-globulin or HBIG in a drug-using population where multiple HBV exposures are likely, the results observed in most trials are not comparable to hepatitis B associated with drug abuse in the military. Therefore, before a recommendation for use of routine γ-globulin or HBIG can be made for drug-related hepatitis in the military, efficacy of standard γ-globulin and/or HBIG should be demonstrated in this population.


[Immunocytogenesis and formation of antibodies in experimentally aged animals (author's transl)]. The authors use white rats with experimentally induced Progeria-like syndrome of Selye. The immunologic response following inoculation with the extracellular microbe-parasite D, pneumoniae and the intracellular parasite Br. abortus 19 is studied serologically, cellularly and biochemically. The experimentally aged rats show diminished and retarded plasmocyte reaction and weaker and slower agglutinine synthesis as compared with the controls. The immunochemical analysis of the serums of the experimentally aged rats reveals changes in the serum fractions and an affected ratio of albumin/globulin. The data suggest a disturbed adaptation and a reversal to an older ontogenic form of the aged organism.


[Silicosis and scleroderma--attempt at synopsis]. Progressive systemic sclerosis obviously more frequently appears in men working under silica dust exposure than in professionally not selected groups. Moreover, silicosis and sclerosis frequently occur combined. Pathogenetic association with regard to immunologic patterns may give an explanation for this phenomenon. Sclerosis occuring in the course of silicosis ought to be included when impairment for compensation purposes is taken into consideration.


[Radiotherapy of hematologic diseases].

[Radiotherapy of hematologic diseases]. Possibilities and limits of radiotherapy are described in a survey in the following diseases: Undifferentiated cell leucoses: The radiotherapy of the central nervous system brings an increase of the 5-year survival rates, since the localisations in this region existing in 45-70% are only insufficiently reached by the cytostatic substances because of the defective blood-liquor passage. On the other hand, other indications to radiotherapy recede into the background. Chronic leucoses: In the foreground of the application is the ray-therapy of the spleen, of which, apart from the local effect on the splenic tumour, also an improvement of the remission rates is expected. At adequate indication also the ray-therapy of infiltrates in the lymph nodes and other localisations achieves good palliative results. The extra-corporeal irradiation of blood is a method, the usability of which must still be proved. With the introduction of the modern cytostatic drugs the exposure of the whole body has lost significance. Lymphogranulomatosis: In this disease the radiotherapy has caused a decisive change: In these cases the recognition of the local development of the disease and its continuous spreading was decisive, and issuing from this also the simultaneous irradiation of the defluxion areas and the application of an oncolytic dose. According to the stage with or without combination of cytostatic drugs an exact plan of therapy is made. Here healing rates of 70-80% in stage I are to be expected. Myeloma: Here the radiotherpy has palliative tasks, with correct indication good effects are to be expected. Polycythaemia vera: In this disease radiotherapy in form of incorporation of radioactive phosphorus is the remedy of choice.


[Therapeutic angiography].

[Therapeutic angiography]. In arterial or venous hemorrhages of the upper gastrointestinal tract, pharmacotherapy and catheter embolization are alternative methods if conservative treatment proves ineffective and surgery offers no prospect of success. A stop in the acute arterial or venous hemorrhage from the upper gastrointestinal tract -- even though only temporary in most instances - may thus be achieved. The choice of one of the two methods and the result depends on the location of the hemorrhage, the age of the patient and the coagulation qualities of the blood. Experience in this respect is quite new, experience which - in spite of all criticism - justifies a modest amount of optimism particularly in view of the desparate situations one is frequently confronted with in acute gastrointestinal hemorrhage.


Cell-mediated immunity in lumphocytic choriomeningitis. I. The specificity of the cytotoxic T lymphocytes. The specificity of cytotoxic T lymphocytes (CTL) generated during murine lymphocytic choriomeningitis (LCM) has been investigated. CTL were obtained from the spleens of mice injected i.p. with LCM virus. The cytotoxic activity of the CTL was tested in an in vitro 51Cr cytotoxicity assay using infected macrophages or fibroblasts as target cells. At the peak of the cytotoxic T cell response (7-8 days after infection) the cytotoxic action was restricted to syngeneic virus-infected target cells. Using H-2 recombinant mice the target antigen of the CTL generated could be identified as products coded for by either the H-2 K or H-2 D region of the major histocompatibility complex. I region identity between CTL and infected target cells was insufficient for optimal lysis to occur. During the early phase of LCM virus infection there was a transient phase during which non-infected H-2 histocompatible targets were lysed as efficiently as virus-infected target cells. This finding may suggest, that during the early phase of LCM disease self-reactive cytotoxic T lymphocytes are temporarily present in LCM virus-infected mice.


[Studies on the mechanism of L-agglutination (author's transl)]. The mechanism of L-agglutination (LA) was studied by fractionating human sera by means of Sephadex G 200 gel filtration. LA was found to be mediated by 2 factors: the rheumatoid factor (RF) as a macroglobulin, occuring only in sera from patients with rhematoid arthritis, and an IgG globulin, found also in normal sera, probably a highly prevalent incomplete streptococcal antibody. RF-containing IgM fractions, by themselves devoid of LA activity, can be rendered positive not only by IgG from the same serum, but also by IgG from normal sera or by commercial human IgG. After absorption of sera containing the RF with streptococci LA disappears; by addition of normal serum, IgG from normal serum or  human IgG LA titers were attained similar to those of the respective native serum. A treatment of RF-containing sera with mercaptoethanol leads to a largely parallel disapperance of LA and latex fixation. Three substrains of Streptococcus pyogenes SF 130 (type) 1), Varying in their endowment with type-specific antigens, reacted similarly in LA.


Effects of maternal immunization on the survival and postnatal development of the offspring. Immunization of females prior to mating altered the size of their litters and the incidence of postnatal death and runting, and the effect varied with the antigen used. Litter size was decreased after immunization with an aggregated synthetic polypeptide or with DNP-BGG, but not with aggregated insulin. Postnatal mortality was increased after immunization with an aggregated polypeptide, aggregated insulin or DNP-BGG. Postnatal runting was increased after immunization with the aggregated polypeptide or with aggregated insulin.


Cell-mediated cytotoxicity against Sendai-virus-infected cells. After injection of Sendai virus, a parainfluenza virus type I, mice generate cytotoxic lymphocytes which lyse specifically Sendai-virus-infected target cells in vitro. Their action is not inhibited by specific antibody in vitro. Killer cell activity appears 4 days after infection, reaches a maximum on the 7th day and disappears on the 14th to 16th day. Decrease of cytotoxic cell activity is correlated with an increase of haemagglutinating antibodies. The cytotoxic effector cell could be characterized as a thymus-derived cell, there is no specific activity in antibody-dependent cell-mediated cytolysis (ADCC). The degree of cytotoxic effector cell activity is only slightly influenced by the dose of injected infective virus. Using different syngeneic Sendai-virus-infected cells as targets for cell-mediated cytotoxicity, a tumor line was not lysed by cytotoxic lymphocytes in spite of viral surface antigens. Preliminary experiments were performed to demonstrate the H-2 gene restriction of the cytotoxic interaction. Using macrophages and tumor cells as targets only syngeneic infected target cells were lysed.


Mixed leukocyte reaction in rats. Effect of immunization with bovine encephalitogenic protein and Freund's complete adjuvant. Rats of certain strains immunized with bovine encephalitogenic protein (EP) in Freund's complete adjuvant develop an impairment of the mixed leukocyte reaction (MLR) similar to that seen in rabbits with experimental autoimmune encephalomyelitis and in humans with certain diseases, including multiple sclerosis. The rats mount a cell-bound response to EP, but encephalomyelitis does not develop. The component causing the impairment was analysed in a culture system using inbred rat strains and F1 hybrids, thoracic duct cells as a source of lymphocytes, and blood as a supplement to the cultures. In normal rats, it was shown that the effects of responding and of stimulating lymphocytes could be separated and that the supportive action of the added blood was probably due to macrophages (monocytes); also that the added blood could in many experiments be replaced with 2-mercaptoethanol (2-ME). The impairment present in immunized rats is at least largely due to a defective supportive activity of the blood (monocytes) and can be restored with 2-ME. The results argue that the MLR impairment seen in immunized rats is due to a faulty macrophage function.


[The cervical cycle as shown on the example of morphological examinations in pregnancy with special reference to subcylindrical vacuolization]. Cyclic alterations of the endometrium are known for certain, but similar processes of the mucous membrane of the uterine cervix are vague. In pregancy there is a considerable increase in secretion. The epithelium of surface and glands becomes stratified and the nuclei of cylindrical epithelial cells withdraw from the base of the cells and often originate retronuclear vacuols. From these vacuols one has to distinguish so-called subcylindrical vacuols which only appear during pregnancy. They are seen between epithelium and basal membrane. Electron microscopical investigations show the vacuols lined partly by atrophical partly bysecretory cylindrical cells. Histochemically the contents of the vacuols are similar to the cytoplasm of cylindrical cells and the contents of glands. These alterations of the cervical mucous membrane are dependent on hormonal stimulation and in conclusion it must be possible to demonstrate the hormonal situation andits disturbances by investigations of the mucous membrane of the cervix uteri.


Inhibition of electron transport on the oxygen-evolving side of photosystem II by an antiserum to a polypeptide isolated from the thylakoid membrane. A polypeptide fraction with the apparent molecular weight 11 000 was isolated from stroma-freed chloroplasts from Anthirrhinum majus. An antiserum to this polypeptide fraction inhibits photosynthetic electron transport in chloroplasts from Nicotiana tabacum. The relative degree of inhibition is pH dependent and has its maximum at pH 7.4. The maximal inhibition observed was 93%. The dependence of the inhibition on the amount of antiserum yields a sigmoidal curve which hints at a cooperative effect. A calculation of the Hill interaction coefficient gave the value of 10. The inhibition occurs on the water splitting side of photosystem II between the sites of electron donation of tetramethyl benzidine and diphenylcarbazide. Tetramethyl benzidine donates its electrons before the site where diphenylcarbazide feeds in its electrons. Analysis of the steady state level of the variable fluorescence also indicates that the inhibition site is on the water splitting side of photosystem II. Tris-washed chloroplasts are equally inhibited by the antiserum and the inhibition is also observed in the presence of an inhibitor of photophosphorylation like dicyclohexyl carbodiimide and in the presence of the uncoupler carbonylcyanide m-chlorophenyl hydrazone (CCCP) which means that the inhibitory action is directed towards the electron transport chain. Valinomycin which is supposed to affect the cation permeability of the thylakoid membrane has no influence on the inhibitory action of the antiserum. The same is valid for gramicidin. Methylamine on the other hand can induce a state in the thylakoids in which the antiserum is not effective. If the antibodies are already adsorbed prior to the methylamine addition then the high inhibitory effect by the antiserum remains unchanged upon addition of methylamine. From the experiments it follows that a component from the vicinity of photosystem II is accessible to antibodies that is, the component is located in the outer surface of the thylakoid membrane. It appears that the inhibitory effect is produced in the course of the light reaction.


[Live enteric typhoid vaccine consisting of a Vi-negative double-dependent mutant of S. typhi and a Vi-positive strain of Citrobacter]. The authors present experimental data on the study of the living enteral vaccine against the typhoid infection from the Vi-negative strain of salmonella with a double-dependence by streptomycin and purine, and from the Vi-positive strain -- citrobacter 5396/38. The method of immunoelectrophoretic analysis showed an indenticity of the O- and H-antigens of the doubledependent mutant with the O- and H-antigens of the typhoid strains of bacteria (Ty2-4446 and 5501). A sufficiently marked immunological reaction was revealed in the tests of antibody formation and in the study of the preventive activity of the sera of the immunized rabbits. The efficacy of the enteral immunization with the associated vaccine consisting of a doubledependent mutant of typhoid bacilli and the citrobacter strain in the doses tested (a 6-fold immunization) was demonstrated in experiments on albino mice. Association in one preparation of the cultures under study did not lead to any changes in the immunogenic properties of these strains. The cells of the mutant strain administered per os gave a positive culture (from the mouse organism) only in the course of the first 24 hours, in difference from the citrobacter strain which gave a positive culture in the course of 14 days.


[Genetic control of the synthesis of Newcastle shigella type-specific VI antigen]. The authors studied the localization of a determinant of the VI type antigen, Sh. newcastle being used in the capacity of donors and E. coli K12 as a recipient. The determinant of the VI type-specific antigen was apparently controlled by the chromosomal gene localized in the region of the histidine operon (on the left from it; 45% of linkage with the his marker).


[Antigenic composition of different forms of Sh. sonnei]. Data are presented on the study, with the aid of immunophoresis, of the antigenic composition of the I, II phases and R-forms of Sh. sonnei. From 5 to 7 antigens differing by thermoresistance, electrophoretic and diffuse mobility were found in the composition of microbial cells of various forms Sh. sonnei. Differences between the I phase on the one hand and cells of the II phase and R-form, on the other hand, consisted both in the quantitative composition of components of their thermostable O-antigens and in the structure of specific lipopolysaccharides of the forms.


[Protective properties of IgM in experiments on animals]. On a model of intraperitoneal infection of albino mice the authors demonstrated a protective action of the fraction enriched with IgM and of gamma-globulin isolated from the normal human blood serum, against E. coli O111. The intensity of the protective action depended on the method, duration of administration of the preparation and also on the infective dose. Protective properties of the fraction enriched with IgM were more pronounced in comparison with the gamma-globulin preparation.


[Effectiveness of specific treatment of rabies. II. Analysis of cases of human death from rabies following a complete course of treatment with specific agents]. Of 79318 persons treated of rabies in the Uzbek SSR in 1968--1974 there were recorded 16 cases of death of clinically manifest rabies. The course of treatment included injection of rabies gamma-globulin in a dose of 0.5 ml per 1 kg of body weight and the maximal dose of rabies vaccine. The incubation period lasted from 44 to 833 days (229.6 on the average). The authors considered that irrational use of high doses of rabies preparations was the cause of unsuccessful treatment of rabies.


[Epidemiologic and clinical features of infectious hepatitis in children against a background of massive prevention of the disease with microdoses of gamma-globulin]. The authors analyzed epidemiological and clinical peculiarities of infectious hepatitis in children aged under 11 years in conditions of mass-gamma-globulin prophylaxis in microdoses the last 5 years in Voronezh. It appeared that the incidence of the disease fell and that its clinical course became milder. In connection with reduction of the incidence of infectious hepatitis in the age group of under 10 years, the incidence of the disease was relatively higher in children aged from 10 to 14 years, with a tendency to levelling-out the periodic autumno-winter elevations among preschool children. Introduction of decreased gamma-globulin dose for hepatitis prophylaxis led to a lesser expenditure of the preparation and thus permitted to vaccinate more children.


[Isoelectric spectra of hepatitis B antigen antibodies to it]. Isoelectric focussing in a stable pH-gradient of ampholines, serum and fraction 4-4, containing hepatitis B antigen demonstrated that the antigen occupied the pI 4.31--6.03 zone with the maximum activity at pI 4.31--5.56. The activity of antibodies to the hepatitis B antigen was revealed in a donor serum and in the gamma-globulin fraction in the pI 7.76--8.4 zone with the activity maximum at pI 8.03--8.23. It was demonstrated immunochemically that antibodies against hepatitis B antigen belonged to immunoglobulin G.


[Neurotropic activity of the gamma-globulin fraction in the serum of schizophrenic patients]. The report is concerned with the influence of the blood sera of schizophrenic patients on the electric activity of neurons in snails and its 3 fractions containing respectively nonprotein compounds, gamma-globulins and the remaining proteins. It was established that the amount of neurotropic activity only of gamma-globulin fractions do not differ significantly from the respective figures of the blood sera of schizophrenic patients. In this connection the authors come to the conclusion of a possible immune nature of the neurotropic factor in the blood sera of schizophrenic patients.


Cytophotometry of feulgen-naphtol yellow S stained liver cells--a computerized method for the calculation of nuclear protein. A computerized method was developed to calculate the nuclear protein content of Feulgen-Naphtol Yellow S stained liver cells from cytophotometric data. After two consecutive scannings at 570 nm (DNA) and 435 nm (protein), the nuclear localization and shape are defined and nuclear protein is calculated by a method which corrects for the nuclear-cytoplasmic overlap at the nuclear periphery. The results obtained by the procedure are highly reproducible and are in accordance with the results of biochemical determinations of nuclear and total cell protein reported by others.


Endocrine function in patients with suprasellar and hypothalamic tumours. The pituitary function in 35 consecutive patients with a non-pituitary suprasellar tumour was studied. In craniopharyngiomas and hypothalmic gliomas severe hormonal disturbances were the rule. In suprasellar cysts and meningiomas and in optic nerve gliomas the hormonal disturbances were on the whole less marked and related to the size of the tumour as judged from the pneumoencephalography, larger tumours leading to more extensive alterations than smaller neoplasms.


Cytology of serous effusions: a comparative study of two slightly different preparative methods. A prospective study of 100 serous effusions was made by two slightly different technics of smearing cells from the centrifuged sediment onto glass slides. Those made with the cotton swab had greater cell concentration and better cell morphology than matching slides prepared by the pipette-"hematologic" two-slide technic. Exclusive use of the cotton swab, therefore, is recommended in making these preparations. On follow-up, in this series of 72 patients, there was a false positive rate of zero per cent, and a false negative rate of 29%.


Calcium and stimulus secretion coupling in the neurohypophysis. V. The effects of the Ca2+ ionophores A23187 and X537 A on vasopressin release and 45Ca2+ efflux; interactions with sodium and a verapamil analogue (D600). Slices from ox neurohypophyses were incubated in a calcium-free medium with the ionophores A23187 or X537A. X537A (5 X 10(-5) mol/l) caused a marked release of vasopressin, neurophysin and protein to the medium. A23187 (2 X 10(-5) mol/l) did not cause any release by itself, but when Ca2+ was added to the medium in the presence of the ionophore, an increase in the release of vasopressin, neuorphysin and protein occurred. Release of lactate dehydrogenase and peptidase were not affected by the ionophores. The secretion caused by A23187 was abolished by D600 (a verapamil analogue) (2 X 10(-5) mol/l) whereas the effect of X537A was unchanged. The effects of X537A were strongly inhibited by removal of sodium from the medium. Re-addition of sodium to the medium caused a marked release. Gramicidin (10(-6) or 5 X 10(-5) mol/l) had no effect on secretion. Efflux of 45Ca2+ from pre-loaded slices was drastically reduced in a sodium-free medium. X537A caused an increase in the efflux rate of 45Ca2+ both in medium with a normal concentration of sodium and when slices had been incubated in a sodium-free medium. A23187 and X537A both released 45Ca2+ from a neurohypophyseal mitochondrial fraction. When sodium in a concentration of 20 mmol/l was added to this fraction, the Ca2+ accumulation was inhibited. This effect was reduced by inorganic phosphate up to a concentration of 2 mmol/l.


Sputum fixatives: how safe is 50% alcohol? It is believed that the following conclusions can be drawn from this study. 1) Fifty per cent ethyl alcohol fixative is bactericidal to 90 to 95% of the bacterial organisms present in the specimens submitted to cytology. 2) The fixative definitely appears to be selective against gram-negative organisms, both cocci and rods. 3) The fixative does not alter the ability of the organisms to take a characteristic stain. 4) The fixative does appear to alter the ability of the Histoplasma and the Cryptococcus to grow after a two-hour minimum fixation. No growth was noted after six weeks.


Interrelations between ABO blood group, plasminogen, alpha1-antitrypsin, alpha2-macroglobulin and the platelet count in blood donors. The level of plasminogen alpha1-antitrypsin, alpha2-macroglobulin and the platelet count were measured in 511 blood donors. The mean level of alpha1-antitrypsin was significantly lower in men of group B compared with that of group O. No other differences between the blood groups reached statistical significance. Woman had higher mean levels of alpha1-antitrypsin and alpha2-macroglobulin and a higher platelet count than men. The levels of plasminogen and alpha1-antitrypsin were significantly higher in women using oral contraceptive compared with those who were not. The level of alpha2-marcoglobulin fell with age until the 60-64 year age-group in men and the 40-49 year age-group in women. A positive correlation existed between the alpha1-antitrypsin and the alpha2-macroglobulin level and between the platelet count and the plasminogen level.


[A combined calcium--fat staining with determination of vessel wall permeability]. A simple combined method for staining the arteries of fetuses and newborns with following demonstration of the vessel wall permeability for macroscopic and microscopic investigations is described.


Calcitonin cells and unusual follicles of the thyroid of the indian grey mongoose, Herpestes edwardsi (Geoffroy). Histological preparations of thyroid, parathyroid and thymus glands of Herpestes edwardsi were examined for calcitonin cells. They reveal that (1) the thyroid calcitonin cells are oval, rounded and rarely elongated in shape; these cells and their nuclei are distinctly larger than those of the follicular cells and their nuclei; (2) calcitonin cells are unevenly distributed in the thyroid, with the result that certain portions of the thyroid are completely devoid of these cells; (3) on an average, calcitonin cells are in a ratio of 10-15 cells/100 follicular cells; (4) the parathyroid and thymus glands do not display calcitonin cells, and (5) the thyroid gland displays unusual follicles of two categories, (a) follicles with ciliated epithelial cells and (b) follicles with squamous epithelium.


Effect of sympathetic denervation on chronic cold injuries in rabbit brain. A microangiographical and histological study. Sympathetic denervation of the brain vessels was performed uni- or bilaterally in 13 adult rabbits, and cortical cold lesions were induced bilaterally 10-18 days afterwards. After a further 4-35 days the vascular and morphological changes in the lesion areas were studied histologically and with angiography. No difference was observed between the denervated and the non-denervated lesion areas. The results are discussed with regard to a proposed functional plasticity of the chronically denervated brain vessels and/or to possible changes in metabolic demands of the brain tissue.


Insecticides in human breast milk.

Insecticides in human breast milk. Fifty samples of human breast milk were analysed by gas chromatography and thin layer chromatography for DDT (dichlordiphenyltrichloraethan), hexachlorobenzol, benezene-hexachlorides, dieldrin, aldrin and heptachlorepoxide. The three first substances were found in all samples in amounts varying from quite small up to eleven times the WHO recommended maximum for cow milk. The other substances were found in fewer milk samples, however, in some of these samples they were found in relatively high amounts. In Norway, only DDT has been investigated earlier (4), and compared to that study, there has been no significant change in the mean concentration of DDT in human breast milk during the last five years. The insecticide content was highest in colostrum and decreased with increasing duration of lactation. Considerable fluctuations in the content of insecticides were recorded in repeated milk samples collected from the same woman a few days apart. The percentage of samples with insecticide content higher than that permitted for cow's milk was greater in May/June (79%) than in early April (54%). The significance of these findings is discussed.


Serum alpha-fetoprotein levels in extrahepatic biliary atresia, idiopathic neonatal hepatitis and alpha-1-antitrypsin deficiency (PiZ). Serum alpha-fetoprotein levels were measured using a sensitive radioimmunoassay in 77 infants presenting with persistent conjugated hyperbilirubinaemia. A breed range of alpha-fetoprotein concentrations occurred in both the 23 infants with extrahepatic biliary atresia and the 35 with idiopathic neonatal hepatitis but the 13 with alpha-1-antitrypsin deficiency had uniformly low levels. High alpha-fetoprotein concentrations (above 10 000 mug/1) favoured the diagnosis of neonatal hepatitis especially in the first ten weeks of life, but the overlap between neonatal hepatitis and extrahepatic biliary atresia was large and alpha-fetoprotein determination cannot be recommended as a reliable method for distinguishing the two conditions. Serial alpha-fetoprotein values showed no consistent relationship with standard liver function tests and gave no guide to prognosis. There was an association between alpha-fetoprotein production and needle biopsy evidence of hepatic giant cell transformation. The uniformly low alpha-fetoprotein levels in alpha-1-antitrypsin deficient infants with neonatal hepatitis is a new observation and possible mechanisms for disordered glycoprotein release are discussed.


The vaccinia virus haemagglutinin: enzyme sensitivity and some physiocochemical properties. Purified vaccina virus haemagglutinin (VHA) was found to be sensitive to trypsin both with regard to haemagglutinating capacity and antigenic property. Phospholipase C and lipase had no effect on the haemagglutination (HA). Both the HA and the antigenic property thus depend on the integrity of the protein part. The loss of HA after treatment with trypsin may be due to breakdown of an essential protein or to secondary rearrangement of the whole structure, including the lipid part. Cells infected with an HA negative mutant of vaccinia virus also lacked the antigenic part of VHA. The sedimentation constant of VHA from HeLa cells was about 50.


The fine structure of cells of Trichomonas vaginalis Donné obtained from the exponential phase of growth and from stationary cultures. In cells of Trichomonas vaginalis in logarithmic phase of growth (log. T. vaginalis) the number of free cytoplasmic ribosomes was higher than that in cells from stationary cultures (stat. T. vaginalis). Polyribosomes closely surrounding the chromatic granules were present in equal amounts in both categories fo cells, which indicates that they have specific functions and are not directly involved in the protein synthesis of the growing cell. The volume of the Golgi region and of the food vacules were approximately twice as large in log. T. vaginalis cells as in stat. T. vaginalis cells. The round and slightly ovoid chromatic granules were smaller in log. T. vaginalis cells than in stat. T. vaginalis cells, whereas elongated granules were more numerous in the former than in the latter cell type. The elongated chromatic granules presented a coarser texture than ordinary round granules. The chromatic granules probably replicate by binary fission a process which seem to follow an internal reorganization of the granules. Polysaccharides were demonstrated on the cell surface and on the membranes of the Golgi and endocytotic vesicles if sectioned cells were stained with phosphotungstic acid after inert dehydration and embedding in a water-missible methacrylate. Ruthenium red and colloidal iron only showed affinity for the filamentous cell coat.


Studies of the antigenic structure of Trichophyton rubrum, Trichophyton mentagrophytes, Microsporon canis, and Epidermophyton floccosum by crossed immuno-electrophoresis. By means of crossed immuno-electrophoresis, 35 antigens of T. rubrum, 26 of T. mentagrophytes, 35 of M. canis, and 25 of E. floccosum have been demonstrated. Tandem crossed immuno-electrophoresis has revealed two antigens common to T. rubrum and the other species, which, however, differ within the species. No common antigens have been demonstrated between T. mentagrophytes, M. canis, and E. floccosum. The technique for preparation of extracts and production of immune serum is described, and the procedure for the electrophoresis is outlined.


Statistical analysis of the different factors which could affect postextrasystolic potentiation in the human heart. The effect of different coupling indices and intervals that could theoretically affect postextrasystolic potentiation has been investigated. A total of 150 ventricular premature beats corresponding to 20 patients submitted to routine cardiac catheterization were studied. Only single ventricular premature contractions following at least four regular sinus beats were considered. Percentage changes in left ventricular systolic pressure, end-diastolic pressure, and max dp/dt were correlated against seven indices and intervals. Index 2 (coupling interval/coupling interval + postextrasystolic pause) gave the better correlations. Besides, this Index includes two intervals that were demonstrated to have statistical significance when individually considered. It has been proved that in the first postextrasystolic beat the highest values of max dt/dt, or left ventricular systolic pressure occurred in early ventricular premature beats, giving a negative regression with Index 2, while in the second postextrasystolic beat the highest values of max dp/dt and left ventricular systolic pressure corresponded to late prematuring beats, giving therefore positive regressions with Index 2 (slope inversion phenomenon). The third and fourth postextrasystolic beats had similar positive regressions but with progressively smaller slopes. Correlations between left ventricular end-diastolic pressure and Index 2 were very poor. It is suggested that variations in baroreceptor activity could account for the different forms of potentiation observed in early and late extrasystoles. In five cases, there were no consistent differences in potentiation when premature beats were elicited from either right or left ventricles.


Minimal heparin cofactor activity in disseminated intravascular coagulation and cirrhosis. An assay technic for measuring heparin cofactor activity in which antithrombin activity can be assessed without plasma attenuation even in the presence of therapeutic levels of heparin is presented. Heparin-activated anti-thrombin activity was markedly depressed in plasmas of four patients with disseminated intravascular coagulation and in ten patients with cirrhosis. Residual activity in those plasmas appeared qualitatively normal, and no inhibitor (platelet factor IV activity) was observed. Plasmas from patients with disseminated intravascular coagulation and cirrhosis required more heparin to obtain in vitro clotting time prolongation equivalent to normal.


Lupus erythematosus cell preparation-antinuclear factor incongruity. A review of diagnostic tests for systemic lupus erythematosus. The cases of 20 patients, each of whom has a positive lupus erythematosus cell preparation and a negative antinuclear factor test, are presented. The concept of a false-positive lupus erythematosus preparation is suggested. Five common mechanisms causing a false-negative antinuclear factor test are discussed and evaluated. Clinical material from the 20 patients is described and pitfalls in diagnosing systemic lupus erythematosus are reviewed.


Positive controls for fungal detection in tissue sections. Lyophilized Histoplasma capsulatum, Coccidioides immitis, Blastomyces dermatitidis and Aspergillus fumigatus are rehydrated, evenly dispersed, and made into smears using a bacteriological loop and alcohol-cleaned slides. Smears are air dried for 15 minutes, fixed in concentrated formaldehyde fumes for 30 minutes, postfixed for one hour in ten per cent buffered formaldehyde, and stored at room temperature. These preparations are used as positive controls for the Grocott stain as recommended for confirmation of fungi detected in tissue sections.


Automated hematoxylin and eosin staining for large volumes of tissue. In order to facilitate the large volume of hematoxylin and eosin (H & E) slides generated by 18 experiments from various programs at the National Center for Toxicological Research, a rapid staining method became imperative. Automated staining with the Gam Rad Stainomatic was decided upon using Gill's no. 1 hematoxylin with a 30-minute staining schedule. Presently, 1,080 slides per instrument per day are stained with a good dependable stain. The cost of this H & E staining method is 0.15 cents per slide.


Immune complex receptors on cell surfaces. II. Cytochemical evaluation of their abundance on different immune cells: distribution, uptake, and regeneration. A recently developed method for ultrastructural demonstration of cell surface receptors for immune complexes is applied to evaluation of these receptors on various cell types. The method entailing incubation with a complex of horesradish peroxidase (HRP) and antibody to HRP (anti-HRP) disclosed dense foci indicative of immune complex receptors distributed at 30- to 120-mmu intervals over macrophage surfaces. Invaginations, loop-like evaginations, and pinocytotic vasicles stained prominently. The number of stained immune complex receptors averaged 200,000 per oil-induced macrophage and 120,000 per noninduced macrophage, as determined from counts of focal deposits in electron micrographs. Receptor periodicity on giant cells present in oil-induced exudates resembled that on macrophages, but the larger giant cells contained an estimated 1.5 million sites. Although receptor periodicity on eosinophils and neutrophils equaled that on macrophages, the staining was lighter and was interrupted by intervals of unstained membrane. Neutrophils averaged 28,000 and eosinophils 35,000 receptors per cell, whereas those lymphocytes with receptors averaged 3,500 per cell. Viable cells incubated with anti-HRP sequentially exhibited about half as many reactive sites as did cells incubated with immune complex. When warmed to 37 C, viable macrophages and eosinophils pinocytosed soluble immune complexes almost completely within 30 minutes and phagocytosed insoluble complexes more slowly. The endocytosed soluble immune complexes were sequestered within tubulovesicular structures in addition to the expected phagocytic vacuoles. Receptors appeared fully active on macrophages that were restained with soluble, cold immune complex after they had endocytosed immune complex in the course of a 30-minute warming interval.


The rapid identification of "normal" parathyroid glands by the presence of intracellular fat. Staining of frozen sections of small parathyroid biopsies from patients with primary hyperparathyroidism with Sudan II or IV revealed the almost uniform presence of numerous prominent intracellular sudanophilic bodies in the chief cells of suppressed "normal" parathyroid glands. These sudanophilic bodies were generally absent from the abnormal chief cells of parathyroid adenomas and chief cell hyperplasias. This difference in intracellular lipid provides a rapid, reliable, and easy method for distinguishing at the time of parathyroid exploration between an adenoma which is always accompanied by "normal" parathyroid glands and a chief cell hyperplasia in which all of the glands are abnormal.


Pathophysiology of enlargement of the small bowel fold. Enlargement of the valvulae conniventes is an integral part of the pattern diagnosis of primary small bowel disease causing malabsorption. The pathophysiology underlying enlargement of the fold and the most typical diseases with prominent folds leading to malabsorption are discussed. Differential diagnosis is based on enlargement of the fold and on secondary signs.


Evaluation of hepatoma chemotherapy by alpha-fetoprotein determination. Effects of chemotherapy on twenty-nine histologically proved and alpha-fetoprotein (AFP) -positive hepatomas were evaluated by changes in serum AFP levels according to the criteria of our previous report. By transfemoral local infusion of mitomycin C, six of eight patients showed more than 50 per cent decrease of AFP levels and regression of the tumor, and the others showed fair effects. By single shot infusion, 50 per cent decrease of serum AFP levels and tumor regression were noted only in three of thirteen patients, and chemotherapy by systemic infusion in eight patients was much less effective. As evaluated by AFP levels, histologically well or moderately differentiated hepatocellular carcinomas responded better to chemotherapy than poorly differentiated ones.


Reaction to radiographic contrast agents: new developments in etiology. Although a significant amount of histamine is released into plasma during contrast media injections, there still is no evidence to support an immunologic basis underlying contrast media reactions. This study supports previous observations that suggest in vitro leucocyte challenge may be of predictive value in contrast media reactions. Lassar's study in dogs and this study in humans clearly shows that complement activation by alternative pathway occurs during contrast agent administration. However, pathophysiological significance of this observation is not clear.


Evaluation of bovine spermatozoal morphologic features after staining or fixation. Two experiments were conducted to evaluate effects of 3 stains and 2 fixatives on morphologic features of bovine spermatozoa. In experiment 1, the morphologic features of acrosomes of raw and incubated, extended spermatozoa were evaluated after staining with Hancock's Blom's or Wells-Awa's stains or after fixation with buffered glutaraldehyde. Evaluations were done of stained smears by bright field microscopy and of fixed, unstained preparations, by differential interference contrast microscopy, using wet mounts. Raw semen samples from 1st ejaculates of 80 bulls were evaluated. The percentage of spermatozoa with intact acrosomes averaged 83.5% in unstained preparations fixed in glutaraldehyde, compared with averages of 68.1, 74.5, and 67.4% for smears stained with Hancock's, Blom's, or Wells-Awa's procedures (P less than 0.01). From these results, it appeared that procedures for preparing stained smears were detrimental to acrosomes. Although counts for other acrosomal abnormalities differed (P less than 0.01) in each treatment, patterns were inconsistent. With incubated, extended spermatozoa from 57 bulls, glutaraldehyde-fixed, unstained samples had more (55%) intact acrosomes (P less than 0.01) than did samples stained with Hancock's or Blom's procedures (24.0 and 34.7%, respectively, but the former were not significantly different from Wells-Awa-stained smears (49.3% intact acrosomes). In experiment 2, several morphologic characteristics of spermatozoa from 15 1st ejaculates of 7 bulls were evaluated after staining with Hancock's or Blom's stains or after fixation in buffered glutaraldehyde or buffered formal saline fixatives. Higher counts (P less than 0.01) of head abnormalities were found in wet, unstained fixed preparations (4.83, 4.47, 7.87, and 7.93% respectively, for Hancock's, Blom's, glutaraldehyde, and formol saline methods). There were more (P less than 0.05) separated heads on stained, dry smears (1.43, 1.23, 0.47, and 0.47%, respectively, for Hancock's, Blom's, glutaraldehyde, and formol saline procedures). Fixation with buffered glutaraldehyde resulted in higher counts (P less than 0.01) of proximal protoplasmic droplets (2.47, 1.03, 0.67, and 1.43%, respectively, for glutaraldehyde, Hancock's, Blom's, and formol saline procedures). Although not significant, the same trend was observed for distal protoplasmic droplets...