Effects of cyclophosphamide treatment on the in vitro activity of mouse lymphoid cells after nonspecific and specific stimulation. A single injection of 300 mg cyclophosphamide (CY)/kg resulted in the mouse in a reduction of the numbers of lymphocytes. The minimal values for spleen and peripheral lymph nodes (PLN) were about 10% and were reached on day 3. The restoration of the spleen started on day 5, while the number of PLN cells was still low on day 21. CY treatment followed by immunization resulted in accelerated recovery and considerably enhanced cell numbers in both organs. Staining with fluorescent antisera revealed that, although the B cell is the main target cell of CY action, in the spleen the total percentage of B and T cells decreases during the period in which the total cell number increases. Immunization diminished the in vitro responsiveness of spleen and PLN cells on phytohemagglutinin (PHA) and concanavalin A (Con A), 7 and 11 days later. The response of PLN cells on lipopolysaccharide (LPS) was, however, strongly enhanced on day 7. CY pretreatment of normal and immunized mice yielded cell populations which behaved identically on nonspecific stimulation in vitro. Starting 5 days after CY, the PHA and Con A responses for spleen cells were impaired, while the initial low R values for PLN cells showed a steady increase. Stimulation of CY spleen cells with LPS resulted in R values of below 1. Specific in vitro stimulation of lymphocytes of immunized mice was performed with sheep red blood cells and a hapten-carrier complex. CY treatment before immunization resulted in an elimination of the in vitro activity on specific stimulation.


T lymphocytes in atopic children.

T lymphocytes in atopic children. The number of circulating T and B cells and the sensitivity of lymphocytes to stimulation with phytohemagglutinin (PHA), concanavalin A (Con A) and pokeweek mitogen (PWM) was studied in 233 atopic children. The number of T lymphocytes was found to be decreased in cases of rhinoconjunctivitis, asthma as well as atopic eczema. Levels of B lymphocytes were normal. Sensitivity to stimulation with PHA and to a lesser degree, Con A, was significantly decreased whereas stimulation with PWM was unaffected. The severity of the atopic eczema was inversely correlated to T cell numbers. Several lines of evidence indicated that the abnormalities observed were intrinsically associated with the atopic conditions and not evoked by corticosteroid treatment. The results are compatible with the hypothesis that atopy is associated with a defect of a subpopulation of T cells. The possibility that this subpopulation has a suppressor function on reagin formation is discussed.


Allergic contact dermatitis in the B-cell deficient chicken. Allergic contact dermatitis to oxazolone was induced in three chickens rendered B-cell deficient by combined chemical bursectomy with testosterone and cyclophosphamide.


Flash photolysis of enzymes.

Flash photolysis of enzymes. The photoionization of aromatic residues constitutes a major initial photochemical reaction in the flash photolysis of proteins at gamma greater than 250 nm. The ejected electrons have been observed as eaq- and the disulphide bridge electron adduct, and also must be trapped at unidentified sites. The number of tryptophyl (or tyrosyl) residues photo-ionized at 5 musec delay is approximately equal to the number of exposed residues. The flash photolysis data have been related to inactivation by considering how photolysis of these "photolabile" residues can affect enzymic activity, based on the microstructure and available information about permanent alterations and residue specificities. This analysis indicates that hen lysozyme and papain are inactivated by photolysis of an essential Trp residue, that bovine trypsin is inactivated by photolysis of a Trp residue adjacent to the key catalytic Ser and other pathways initiated by excitation of Tyr and Cys, that the efficient photoionization of Tyr and RNase A is not an important inactivating reaction, and that aromatic residues in subtilisn Carlsberg are photosensitive.


Epithelial regeneration of transposed intestine after high doses of X-irradiation. The regeneration capacities of normal and transposed small bowel epithelium were compared in rats after applying high doses of X-irradiation. It has been shown that the potency of the mucosa to regenerate is much higher than assumed and that the mucosa can regenerate after single doses varying from 2000-5000 R. Even in the villus epithelium and in flat epithelium covering infiltrates of the lamina propria cells survive, which are still able to resume proliferative activity several days after irradiation.


[Effect of gamma rays from 60Co on the in vitro incorporation of palmitic acid in lipids of rat liver microsomes]. The gamma-rays of 60Co cause a diminution in the incorporation of palmitic acid in the phosphatidic acids and the neutral fats of rat-liver microsomes. This diminution occurs without morphological change in the microsomal membranes, where the enzymes responsible for this incorporation are situated. Some evidence is offered that lipid peroxides may be involved in this inhibition.


Binding of radiation-induced phenylalanine radicals to DNA: influence on the biological activity of the DNA and on its sensitivity to the induction of breaks by gamma-rays. When an aqueous solution of double-stranded DNA of bacteriophage PM2 containing phenylalanine and saturated with N2O is irradiated with gamma-rays, radiation-induced phenylalanine radicals are bound covalently. Under the conditions used, about 25 phenylalanine molecules may be bound per lethal hit. For single-stranded PM2 DNA, most of the phenylalanine radicals bound are non-lethal. Evidence is presented that, in double-stranded DNA, an appreciable fraction of the single-strand breaks is induced by phenylalanine radicals. Radiation products of phenylalanine and the phenylalanine bound to the DNA decrease the sensitivity of the DNA to the induction of single-strand breaks. There are indications that the high efficiency of protection by radiation products of phenylalanine is due to their positive charge, which will result in a relatively high concentrations fo these compounds in the vicinity of the negatively-charged DNA molecules.


The r.b.e. of different energy neutrons as measured by the heamatopoietic spleen-colony technique. The spleen-colony technique has been used for determining the relative biological effectiveness (r.b.e.) for several energies of neutron radiation. Donor mice were exposed for fission and accelerator-generated neutrons at a variety of doses and energies. Immediately after exposure, donor bone-marrow was removed from the hind legs, and standard amounts were injected intravenously into lethally X-irradiated recipients. After 7 days the recipients spleens were evaluated for surface colonies. Dose-response curves were obtained for each type of radiation and the Do was determined. The neutron r.b.e. values from the Do compared with 250kVp X-rays were: reactor 1.58, 252Cf 1:59, and accelerator varied from 2.85 at 1.0 Mev to 0.85 at 13.4 MeV.


Enzymatic detection of x-ray-induced strand breaks in nuclear DNA on sections of mouse tissue. An enzyme-histochemical method was used to detect X-ray-induced strand breaks in the DNA of mammalian tissue cells. Paraffin sections of ethanol-fixed mouse brain and liver were incubated with three kinds of exogenous DNA polymerizing enzymes, and the amount of in situ incorporation of 3H-deoxyribonucleotides into nuclear DNA was examined by autoradiography. No increase in labelling intensity was observed over nuclei of neurons and astrocytes in cerebral cortex, or over hepatocytes in liver immediately after X-irradiation when compared with unirradiated specimens. In liver Kupffer cells, heavily-labelled nuclei appeared from 30 min to 6 hours after, but were not observed immediately after X-irradiation. This method cannot, therefore, be applied to detect the strand breaks directly induced by X-rays, but it is useful in detecting secondary DNA degradation occurring as a result of nuclear degradation.


Kinetics of nitroxyl radical reactions. A pulse-radiolysis conductivity study. Absolute rate-constants for the reaction of the nitroxyl free radicals TAN and TMPN with radiation-chemically-formed radicals and ions have been determined. k(TAN + X) (in M(-1) sec(-1)=4-0 X 10(9) (for X = OH-), 2-9 X 10(10) (eaq-), 8-0 X 10(9) (H-), 7-2 X 10(8) (-CH2OH), 4-0 X 10(8) (CH3CHOH), 4-3 X 10(8) ((CH3)2COH) 2-8 X 10(8) (-CH2(CH3)2COH), 5-9 X 10(7) (glucose radical), 4-0 X 10(8) (c-C5H9-), and k(TMPN + X)=3-4 X 10(9) (OH-), 7-8 X 10(9) (eq-), 4-9 X 10(9) (H-), 4-4 X 10(8) (-CH2OH), 4-9 X 10(8) (CH3CHOH), 3-6 X 10(8) ((CH3)2COH), 1-5 X 10(8) (-CH2(CH3)2COH), 4-9 X 10(7) (glucose radical), 4-3 X 10(8) (c-C5H9-). Direct measurements by means of a pulse-radiolysis conductivity technique were based on the formation and destruction of charged species in these reactions within certain pH ranges. It is indicated that the radiosensitizing nitroxyls undergo both redox and addition reactions.


The effect of repeated small doses of radiation on recovery from sub-lethal damage by Chinese hamster cells irradiated in the plateau phase of growth. Unfed plateau-phase cells have been irradiated with either single doses or up to ten fractions of X-rays 6 hours apart. The single-dose survival curve had an extrapolation number of 11-4, and the oxygen-enhancement ratio (o.e.r.) was 3-1. Cells were exposed to multiple fractions of 200 rad or 150 rad in air and 600 rad or 450 rad in hypoxia. The resulting survival curves did not fit a multi-target, single-hit model of cell survival, being much steeper than that would predict. The curves were exponential up to five fractions of X-rays, but tended to bend downwards with increasing number of fractions. Cells that had survived five fractions of 200 rad (or 600 rad in hypoxia) 6 hours apart, were less able to absorb damage as sub-lethal than those which had not previously been exposed to radiation. The ratio of the initial slopes of the fractionated survival curves for irradiation in air and hypoxia was 2-1, implying that the o.e.r. "on the shoulder" may be less than that in the exponential region of survival.


Effects of 60Co irradiation on unfed adults and engorged females of the tick Hyalomma anatolicum. Unfed and engorged adults of the cattle tick Hyalomma anatolicum, known to be biological vector of Theileria anmulata in India, were exposed to various levels of gamma-radiation from 0-60 000 R, and the effect on engorgement and redproduction was observed. The radiation tolerance limit for unfed adults is 1000 R for engorgement and reproduction. The sterilizing dose for most of the males is 2000 R but the females can tolerate a slightly higher dose. Engorged females have a tolerance limit of 10 000 R for oviposition and 1000 R for larval development. Higher dosages (20 000-60 000 R) have adverse effects on development, completely inhibiting egg-laying at 60 000 R, but these females (60 000 R) live longer (10 weeks) than those irradiated at lower levels.


Phosphate ester cleavage in ribose-5-phosphate induced by OH radicals in deoxygenated aqueous solution. The effect of Fe(II) and Fe(III) ions. The reaction of OH radicals and H atoms with ribose-5-phosphate (10(-2) M) in deoxygenated aqueous solution at room temperature (dose-rate 2-1 X 10(17) eV/ml-min, dose 5 X 10(18)-15 X 10(18) eV/ml) leads to the following dephosphorylation products (G-values): ribo-pentodialdose 1 (0-2), 2-hydroxy-4-oxoglutaraldehyde 2 (0-06), 5-deoxy-erythro-pentos-4-ulose 3 (0-1) and 3-oxoglutaraldehyde 4 (0-06). In addition, some minor phosphate free products (total G=0-09) are formed. G(inorganic phosphate) =1-3 and G(H2O2)=0-3. On the addition of 10(-3) M (Fe(III) ions, G (1) and G (3) increase to 0-6 and 0-4 respectively. In the presence of 10(-3) M Fe(II), G(1) and G(3) change to 0-4 and 0-8, respectively. The other dephosphorylation products are suppressed by the iron ions. G(1) also increases on the addition of increasing amounts of H2O2. Each product can be assigned a precursor radical formed by hydrogen abstraction from C-5, C-4 or C-3 of the ribose-5-phosphate molecule. Products 1 and 2 are formed by oxydative dephosphorylation of an alpha-phospho radical with preceeding H2O elimination for product 2. Elimination of H3PO4 from a beta-phospho radical leads to product 3; product 4 is formed by elimination of two molecules of H2O from its precursor radical and hydrolytic cleavage of an enol phosphate bond. Deuterium-labelling experiments and the effects of the iron ions and of H2O2 support the mechanisms proposed. The importance of the dephosphorylation mechanisms for the formation of strand breaks in DNA is discussed with special reference to the effects of the radiosensitizers.


Negative pion irradiation of mammalian cells. 1. Survival characteristics of monolayers and spheroids of Chinese hamster lung cells. Monolayers and spheroids of Chinese hamster cells (V79) were subjected to negative pion irradiation under aerobic conditions. R.b.e. values in the pion peak of 1-8 and 1-5 were obtained for monolayers and spheroids, respectively, whereas the r.b.e. for the plateau was found to be slightly higher than 1. In addition, it was observed that the higher resistance of the V79 spheroid cells than the monolayers to gamma-irradiation is not diminished in the pion peak, suggesting that the underlying phenomenon of intercellular communication influences cell survival even after high-LET irradiation.


Cirrhosis and heterozygous alpha1-antitrypsin deficiency in a 4-year-old girl. A 4-year-old girl with hepatic cirrhosis was found to have alpha1-antitrypsin deficiency. The method of FAGERHOL detected that she is a heterozygous carrier of the Pi SZ type. Investigations with the light and the electron microscope and immunofluorescence tests confirmed the diagnosis. The values of alpha1-antitrypsin and trypsin inhibitory capacity gave intermediate values. The fact that in the patient, the Z allele was combined with an S allele may have contributed to the severity of the illness.


[Facial paresis following stapedectomy (author's transl)]. An ususual complication after stapedectomy for otosclerosis is reported. After two vain attempts elsewhere, the tympanic cavity was opened for the third time. Since the incus was missing, a malleovestibuloplasty was undertaken, using the Schuknecht wire prosthesis. Hearing was very good. One day after the operation, a facial paralysis developed, and increased over the following days. After 18 days, the nerve  was found to be oedematous and swollen above the fenestra ovale upto the level of the promontory and had forced the prosthesis out of the fenestra ovale. Six weeks later, nerve function was normal.


Hormonal regulation of gap junction differentiation. Thin-section, tracer, and freeze-cleave experiments on hypophysectomized Rana pipiens larvae reveal that gap junctions form between differentiating ependymoglial cells in response to thyroid hormone. These junctions assemble in large particle-free areas of the plasma membrane known as formation plaques. Between 20 and 40 h after hormone application, formation plaque area increases approximately 26-fold while gap junction area rises about 20-fold. The differentiation of these junctions requires the synthesis of new protein and probably RNA as well. On the basis of inhibitor experiments, it can be reported that formation plaques develop at about 16-20 h after hormone treatment and stages in the construction of gap junctions appear 4-8 h later. These studies suggest that gap junction subunits are synthesized and inserted into formation plaque membrane during the differentiation of the anuran ependymoglial cells.


Topological analysis of the brain stem of the frogs Rana esculenta and Rana catesbeiana. The ventricular sulcal pattern and the cytoarchitectonic organization of the brain stem of the frogs Rana esculenta and Rana catesbeiana have been studied in transversely cut, Nissl stained serial sections. Four longitudinal sulci, the sulcus medianus inferior, the sulcus intermedius ventralis, the sulcus limitans and the sulcus medianus superior could be distinguished in both species. A fifth longitudinal groove, the sulcus intermedius dorsalis, was found only in Rana esculenta. With the aid of the usual cytoarchitectonic criteria 25 cell masses have been delineated in Rana esculenta and 27 in Rana catesbeiana. These cell masses can be distributed over the following categories (numbers added in brackets for Rana catesbeiana, if different from those in Rana esculenta): primary efferent or motor, 8; primary afferent or sensory, 4(6); "relay" centers, 7. Contrary to statements in the literature the reticular formation can be divided into six separate cell groups. The majority of the nuclei form part of the central gray, which constitutes a rather wide zone in anurans; three reticular nuclei lie partly within the stratum griseum and partly within the stratum album; six nuclei are entirely embedded in the stratum album. The morphological pattern of the cell masses and their relationship to the ventricular sulci were studied with the aid of a graphical reconstruction procedure termed topological analysis (cf. Nieuwenhuys, '74 and figs. 15, 16). This analysis yielded the following results: The sulcus limitans extends throughout the rhombencephalon, dividing this brain part into a basal plate and an alar plate. The cell masses in the basal plate fit into two longitudinal zones, a medial area ventralis and a lateral area intermedioventralis. The area ventralis contains three somatic motor nuclei (IV, VI and XII) and the rhombencephalic medial reticular zone. The latter may be primarily considered as a somatic motor coordinating center. The area intermedioventralis contains the visceral motor nuclei of V, VII, IX and X. However, the basal plate also contains a number of non-motor centers, for example the superior olive. The alar plate contains visceral sensory, general somatic sensory and special somatic sensory centers. Two cell masses, the nucleusfasciculi solitarii and the nucleus visceralis secundarius, represent together a discontinuous visceral sensory zone. Both of these nuclei are situated immediately dorsal to the sulcus limitans. The special somatic sensory area, i.e., the area of termination of the eighth nerve, occupies a considerable part of the alar plate. This area comprises, apart from a large zone of diffuse gray, three distinct cell masses...


alpha1-Antitrypsin deficiency and skin abnormalities. A 19-year-old Moroccan male was found to have total absence of serum alpha1-antitrypsin, a major inhibitor of elastase. This patient had chronic obstructive lung disease, hyperextensibility of the skin over the cheeks and wrists, and hyperlaxity of the hand joints. Microscopic sections of the skin revealed a thickened dermis with shortened and rarefied elastic fibers. Ultrastructural study showed collagen fibers with variable and irregular diameters. Elastic fibers were scarce and their relatively poor matrix was surrounded by numberous microfibrils. The outline of the fibers was irregular with deep recesses filled with microfibrils. The ergastoplasm of the fibroblasts was well developed. The differential diagnosis with other connective dystrophies showed the original characteristic of this case. Clinically and histopathologically, the skin abnormalities are probably related to the deficiency in elastase inhibitor.


Preliminary cutaneous pathology observations with the freeze-fracture technique. The freeze-fracture technique was used to study acantholytic cells of untreated pemphigus patients and the cells of histocytosis X. On the membranes of acanthocytosis cells, the particles are randomly distributed and lack the typical desmosome organization, while tight and gap junctions are still found. The study of the cells of histocytosis X provided a better understanding of the structure of the Langerhans granule. The paracrystalline organization of the inner face of the disc-shaped portion does not seem to involve the membranes limiting the granule, but rather to be confined to a periodically arranged, possibly proteic substance contained between them.


Immunobiology of aging.

Immunobiology of aging. Normal immune functions can begin to decline shortly after an individual reaches sexual maturity. Although changes in cellular environment are partially responsible, the decline is due primarily to changes within the cells, especially the T cells and to some extent the stem cells. This is reflected in their inability to proliferate and differentiate efficiently. What needs to be resolved is whether the altered properties of T cells and stem cells are permanent or reversible and, if permanent, whether they are due to a stochastic or genetically programmed event. The decline with age in certain normal immune functions is associated with an increase in the frequency of autoimmune and immune complex diseases, certain types of cancer, and viral and fungal infections. These diseases, compromise normal immune functions in short-lived strains of mice. In long-lived mice and in humans, however, the decline in immune functions to threshold levels seems to predispose in individuals to illness.


Autoimmune disease: the consequence of deficient T-cell function? The age-related decrease in T-lymphocyte function is discussed in relation to the onset of autoimmune disease. It is postulated that the decrease in T-cell "immune surveillance" permits: a) the ingress of viruses whose enzymes modify host glycoproteins and render them immunogenic, and b) the replication of viruses incorporated into the genome of cells during infections in early life. The existence of immunodeficiencies too subtle to be detected by our present laboratory methods is postulated and discussed in relation to diseases observed in geriatric patients. It would seem that the prevention of some geriatric diseases should begin in the pediatrician's office.


Tumor immunology, autoimmunity and aging. Cell-mediated immune function declines with aging, and may be associated with autoimmunity and malignancy. Humoral immune responses also decline with aging. The chief age-related effect on the immune system is a decrease in T-cell function. The "thymus clock" and immunogenesis are discussed in relation to aging. In animals, attempts at immunologic rejuvenation by cellular or hormonal means have not been successful as the results attained by genetic manipulation.


Results of open heart surgery in the septuagenarian. Open heart surgery has been extended to the septuagenarian. At the Long Island Jewish-Hillside Medical Center, 42 patients 70 to 79 years old underwent 43 open heart procedures in the past 7 years. Their average age was 73 years and 62% were male. Eighteen underwent aortic valve replacement; 7 had mitral valve replacement; 18 had coronary bypasses (3 with valve replacement); exploration for pulmonary embolism was done in 1; 1 patient had tricuspid replacement with mitral commissurotomy; 17% (7/42) died during hospitalization or within 30 days of surgery. Mean follow-up is 25 mo., and an additional 19% (8/42) have died at an average of 27 mo. after surgery. Open heart surgery is feasible for the elderly, but at an increased risk when compared to the younger patient submitted to the same procedure.


IgM complex receptors on subpopulations of murine lymphocytes. Murine lymphocytes from spleen, lymph node, and thymus were examined for IgM complex receptors. Lymphocytes from all three organs were found to bind SRBC sensitized with IgM from various sources including: primary anti-SRBC serum, murine and rabbit anti-Escherichia coli LPS sera, and a murine IgM myeloma (MOPC 104E). Rosette formation by lymphocytes with IgM-sensitized SRBC was inhibited by soluble antigen-IgM complexes but not by IgM or antigen alone. Rosette formation was also inhibited by human IgM (Fc)5mu but not by Fab mu. Antiserum and complement treatment of the cells and subsequent recovery of the viable cells by trypsinization, filtration, and washing revealed the IgM rosette-forming cell (RFC) in the thymus to be a T cell. Spleen on the other hand was found to contain both B and T cells capable of binding IgM sensitized SRBC. Removal of both B and T cells from spleen cell suspensions eliminated all IgM RFC. The IgM complex receptor was found to be trypsin insensitive. Anti-Ig column fractionation enriched IgM RFC in spleen and lymph node suspensions passed through the columns, whereas cells bearing surface Ig, IgG complex receptors, and C3 receptors were retained in the columns.


Splenic suppressing factor: purification and characterization of a factor suppressing thymidine incorporation into activated lymphocytes. Suppressing activity upon the mitogen-activated lymphocytes was found in the supernatant (SUP) from the culture of mouse spleen, high-density subpopulation of thymocytes, and peritoneal exudate cells. Suppressing factor was obtained from the non-stimulated lymphocytes cultured for 24 to 36 hr with or without serum. Suppressing activity in the SUP was observed in the incorporation of 3H-thymidine, 3H-uridine, and 3H-leucine into Con A-activated lymphocytes or in the proliferation of L cells. Suppressing factor partially purified by Sephadex G-25 column chromatography was a heat-stable and dialyzable substance(s). Further purification and isolation of this factor by two-dimensional thin layer chromatography revealed that this was thymidine and thymidine monophosphate. The suppression in 3H-thymidine incorporation was attributed to the dilution effect of cold thymidine released from cultured lymphocytes.


Reaginic antibody formation in the mouse. VII. Induction of suppressor T cells for IgE and IgG antibody responses. Intravenous injections of urea-denatured ovalbumin (UD-OA) into OA-primed high responder mice suppressed the antibody response not only to the priming antigen but also to subsequent immunization with dinitrophenyl derivatives of OA (DNP-OA). The transfer of normal spleen cells or OA-primed spleen cells into UD-OA-treated animals did not restore the capacity of responding to DNP-OA to form anti-DNP IgE and IgG antibodies. The transfer of splenic T cell fraction from the UD-OA-treated animals into normal syngeneic mice diminished both IgE and IgG antibody responses of the recipients to DNP-OA. The B cell-rich fraction from the same donors failed to affect the anti-hapten antibody response and enhanced anti-cancer (OA) IgG antibody response of the recipients. It was also found that the transfer of T cell-rich fraction of OA-primed spleen cells failed to suppress antibody response of the recipients to DNP-OA. The results indicated that spleen cells of UD-OA-treated mice contained suppressor T cells which are distinct from helper cells. Suppressive activity of T cells in the UD-OA treated animals was specific for OA. The transfer of the T cell-rich fraction failed to suppress anti-DNP antibody response of the recipients to DNP-KLH.


Enumeration and isolation of rabbit T and B lymphocytes by using antibody-coated erythrocytes. Rosette formation with antibody-coated erythrocytes (Ab-E) was employed for the enumeration and isolation of rabbit B cells (Ig+T-) and T cells (Ig-T+). The cells bearing surface Ig (Ig+ cells) were enumerated by a direct immunocytoadhesion technique utilizing anti-rabbit IgG antibody-coated erythrocytes (Ab-E). To enumerate cells bearing thymus cell antigen (T+ cells), an indirect rosette technique was used in which lymphocytes were first sensitized with guinea pig anti-rabbit thymus cell antiserum and then rosetted with anti-guinea pig IgG Ab-E. To demonstrate the specificity of the anti-thymus cell antiserum, a 51Cr radioimmunoassay for counting rosettes was employed along with visual counting to enumerate Ig+ and T+ cells in lymph node cell populations. When Ig+ and T+ lymph node cells were rosetted simultaneously with sheep and human erythrocytes, no mixed rosettes (less than 1%) were observed. Ficoll-Hypaque gradient centrifugation was used to obtain purified Ig+T- and Ig-T+ cells by removing rosetted T+ and Ig+ cells, respectively. The purity of isolated Ig-T+ cells was indicated by 94 to 95% indirect rosetting with anti-thymus cell antiserum and by 0 to 3% direct rosetting with anti-rabbit IgG Ab-E. The purity of isolated Ig+T- cells was indicated by 90 t0 94% direct rosetting with anti-rabbit IgG Ab-E and by 2 to 3% indirect rosetting with anti-thymus cell antiserum. The percentage of Ig+T- and Ig-T+ cells were determined in peripheral blood and in various lymphoid organs. The isolated Ig+T- and Ig-T+ cells were also characterized by their responses to mitogens. Thus, nearly pure Ig+T- and Ig-T+ cells were isolated by "negative selection,"  which should minimize functional changes of the cells, and thereby facilitate the study of their biologic properties, e.g., their response to mitogens.


The suppressive effect of immunization on the proliferative responses of rat T cells in vitro. II. Abrogation of antigen-induced suppression by selective cytotoxic agents. Rats given large i.v. doses of ovalbumin or sheep erythrocytes manifest suppressed spleen cell responses (3H-thymidine incorporation) to PHA within hours. Removal of glass wool-adherent cells totally restores responsiveness to that of normal nonadherent spleen cell cultures. Carrageenan, selectively toxic for macrophages, partially restores responses of antigen-suppressed spleen cells in culture, suggesting a supportive role for macrophages in the suppression phenomenon. Treatment of donors with low doses of cyclophosphamide (20 to 50 mg/kg) at the time of antigen injection abrogates the ability of their spleen cells to suppress the responses of normal cells to PHA. The low dose of cyclophosphamide required indicates a target other than the B cell or macrophage and suggests the possibility that cyclophosphamide eliminates the suppressor T cell component of the macrophage-T cell complex.


Differentiation of thymocytes: evidence that induction of the surface phenotype requires transcription and translation. Induction of thymocyte differentiation requires transcription of DNA and translation of RNA but not replication of DNA. This was determined in vitro by studying the effects of cytosine arabinoside, hydroxyurea, actinomycin D, camptothecin, cordycepin, cycloheximide, and puromycin on the induction by thymopoietin of thymocyte differentiation, measured by the acquisition of the differentiation allo-antigens TL and Thy-1 by and inducible fraction of mouse spleen cells. This requirement for transcription does not establish whether the new mRNA represents transcripts of structural genes for the thymocyte allo-antigen markers; an answer to this question awaits the results of further experiments.


The significance of varying SRBC/lymphocyte ratio in T cell rosette formation. The incubation ratio of sheep red blood cells (SRBC) to lymphocytes is a critical factor in rosette formation, whereas the length of time SRBC and lymphocytes are incubated together does not significantly affect the percentage of lymphocytes forming rosettes. The graph obtained by plotting percentage of rosette formation against the ratio of SRBC to lymphocytes is similar to that resulting from the formation of bimolecular complexes. If rosette formation is analogous to formation of bimolecular complexes, maximal rosette formation occurs when the system is saturated, i.e., with excess SRBC, and is a measure of the total capacity of a lymphocyte population to form rosettes. In addition, the percentage of rosette formation observed at a limiting SRBC/lymphocyte ratio gives an indication of the avidity of the lymphocytes for SRBC. This interpretation may provide an explanation for the difference between the "active" and "total" rosettes. When the log of the SRBC/lymphocyte ratio is plotted against percentage of rosette formation, a straight line is obtained, suggesting that within a given normal lymphocyte sample, T cell subsets with different avidities are not detected by rosette formation at different SRBC/lymphocyte ratios.


Immunoglobulin-positive mononuclear cells in human peripheral blood: detection by mixed anti-globulin and direct anti-globulin-rosetting reactions. Ig-bearing mononuclear cells were identified in Ficoll-Hypaque preparations of human peripheral blood by using mixed anti-globulin (MAG) and direct anti-globulin rosettes; indicator cells consisted of sheep erythrocytes coated with human F(ab')2 or anti-F(ab')2 antibody, respectively. Of the cell population isolated from 10 normal subjects, a mean of 68% was lymphocytes. However, fewer than 50% of the cells with detectable surface Ig were lymphocytes. On viable cell preparations using chromic chloride-treated sheep erythrocytes (CrCl3SRBC) coated with anti-F(ab')2 antibody, a mean of 20.1% of the lymphocytes formed rosettes, i.e., were B. Up to 6% of peripheral blood lymphocytes formed mixed Ig-rosettes and E-rosettes. On viable lymphocytes using F(ab')2-coated CrCl3SRBC, MAG rosettes were insensitive in detection of B lymphocytes. Formaldehyde treatment of lymphocytes increased the number of B cells detectable to 25.5% of the lymphocyte population. Study of T-enriched and B-enriched populations showed that the observed increase in B cell reactivity was real and not due to MAG-rosetting T cells. A one-stage procedure for T and B lymphocyte separation is described.


Macrophage requirements of CR- and CR+ B lymphocytes for antibody production in vitro. A Sephadex G-10 column coated with antigen-antibody complexes and complement retains complement receptor-bearing (CR+) mouse spleen cells. The effluent is rich in thymus-derived cells (T cells), and contains bone marrow-derived cells (B cells) which carry surface immunoglobulin (Ig), Ir-associated antigen (Ia), and Fc receptors, but no complement receptors (CR-). Although both unfractionated and CR- B cell populations are capable of producing antibody to red cell antigens, they differ in their requirements for the initiation of the response. Unfractionated B cells cooperate with primed as well as unprimed helper T cells; macrophages are required for this cooperation but can be replaced by 2-mercaptoethanol. CR- B cells cooperate with primed but not with unprimed T cells provided macrophages are added to cultures. After addition of culture supernatant from BCG-activated macrophages CR- B cells cooperate with both unprimed and primed T helper cells.


The function of macrophages in antigen recognition by guinea pig T lymphocytes. III. Genetic analysis of the antigens mediating macrophage-T lymphocyte interaction. Activation of immune T lymphocytes by antigen-pulsed macrophages is mediated by the Ia antigens of the guinea pig MHC or the products of closely linked genes. Studies using combinations of macrophages and T cells derived from outbred animals with different Ia antigens and/or Ir gene products have demonstrated that sharing of Ir gene products between macrophage and T cells is not sufficient for effective macrophage-T cell interaction. The role of the Ia antigens in the absence of the linked Ir gene products could not be directly examined because we were unable to identify an animal which bore the full complement of Ia antigens in the absence of the Ir gene that is normally associated with them. The results of these studies support the concept of the functional expression of the Ir gene product in the macrophage.


Evolution of the lymphoid system. I. Evidence for lymphocyte heterogeneity in rainbow trout revealed by the organ distribution of mitogenic responses. Leukocytes from the various lymphoid tissues of rainbow trout (RBT) were tested for their capacity to respond to the lymphocyte mitogens concanavalin A (Con A), lipopolysaccharide (LPS), and purified protein derivative of tuberculin (PPD). Thymocytes responded to Con A but not to LPS or PPD. In contrast, leukocytes from anterior kidney were stimulated with LPS but not with Con A or PPD. Cells from spleen and peripheral blood were stimulated by each mitogen. However, the degree of stimulation at optimally stimulatory concentrations of each mitogen was distinctive. The finding that the patterns of mitogenic responses of cells from each tissue were significantly different suggested that there is lymphoid heterogeneity in the RBT with a unique tissue distribution. The species source of serum utilized as a medium supplement appeared to be capable of markedly affecting mitogenesis. Thus, LPS and PPD stimulation occurred in medium supplemented with rainbow trout serum (RBTS). On the other hand, LPS and PPD stimulation was not observed in medium supplemented with fetal bovine serum (FBS), with the exception of peripheral blood leukocytes which were stimulated by LPS in culture medium supplemented with FBS. Con A stimulated leukocytes from each lymphoid tissue in medium supplemented with RBTS and, with the exception of cells from anterior kidney, also stimulated cells from each tissue in medium supplemented with FBS. The kinetic profiles of the responses of peripheral blood leukocytes to Con A, LPS, and PPD suggested that the extent as well as the time required for maximal stimulation was dependent on the dose of mitogen.


Differentiation of lymphoid cells: B cell as a direct target and T cell as a regulator in lipopolysaccharide-enhanced induction of immunoglobulin production. The cells involved in the stimulatory effect of bacterial lipopolysaccharide (LPS) on the induction of immunoglobulin (Ig) production by rabbit spleen cells cultured in the absence of antigen has been analyzed. Addition of LPS caused a several-fold enhancement of both DNA synthesis and Ig production. These enhanced activities were not significantly affected by depletion of adherent cells in the spleen cell population. Although inactivation of splenic T cells by anti-thymocyte serum (ATS) treatment did not affect the enhancement of DNA synthesis due to LPS, such treatment did adversely affect the enhancement of Ig production by LPS. Furthermore, the enhancement of Ig production of ATS-treated spleen cells by LPS was found to be dependent on the number of thymocytes added. In addition, the prior incubation of ATS-treated spleen cells with LPS resulted in effective enhancement of Ig production when such ATS-treated spleen cells and thymocytes were combined after removal of LPS. An identical experiment, except that thymocytes instead of ATS-treated spleen cells received the prior incubation with LPS, did not result in enhancement of Ig production. Finally, the enhanced Ig production due to LPS was inhibited by hydroxyurea, a known inhibitor of cellular DNA synthesis. The relationship between the mitogenic activity of LPS on B cells, the regulatory function of T cells, and the enhancement of Ig production by LPS is discussed in relation to the contrasting reports concerning the cellular target of LPS.


Structural requirements for arthritogenicity of peptidoglycans from Staphylococcus aureus and Lactobacillus plant arum and analogous synthetic compounds. The comparative studies on the arthritogenicity of chemically well defined peptidoglycans (PG)2 from the cell walls of Staphylococcus aureus (FDA 209P) and Lactobacillus plantarum (ATCC 8014) showed that 1) a polymer of disaccharide-peptide with or without the presence of N-acetylglucosaminyl-ribitol-teichoic acid produced severe arthritis; 2) oligosaccharide-peptides with or without the special structure (N-acetylglucosaminyl-ribitol-teichoic acid in S. aureus, a polymer of rhamnose and glucose in L. plantarum) produced severe disease; 3) disaccharide-heptapeptide-disaccharide with or without the presence of either glucose-ribitol-teichoic acid or a polymer of rhamnose and glucose appeared to be arthritogenic but much less effective for disease production; 4) N-acetylmuramyl-heptapeptide-N-acetylmuramic acid and disaccharide-hexapeptide were non-arthritogenic; 5) none of the synthetic N-acetylmuramyl-peptides, including tetrapeptide, produced the disease. Thus it is concluded that arthritis-inducing activity is related to the peptidoglycan moiety but not to the special structure, and the most important moiety responsible for disease production may be located in a chain length of two or more disaccharide units on PG subunits. However, it is discussed that non-arthritogenicity of peptidoglycans, including synthetic compounds, may be due to a lack of adjuvanticity in rats rather than a lack of antigenic determinant(s) responsible for production of arthritis.


B lymphocyte differentiation induced by lipopolysaccharide. III. Suppression of B cell maturation by anti-mouse immunoglobulin antibodies. Lipopolysaccharide-(LPS) induced differentiation of mouse B lymphocytes to cells synthesizing large amounts of cytoplasmic IgM and IgG2 could be suppressed by antibodies to mu-chains. Maximal inhibition of LPS-induced differentiation was associated with increased cellular proliferation as measured by incorporation of 3H-thymidine, whereas treatment with anti-mu alone over a wide dosage range did not stimulate cellular proliferation. Spleen cells from newborn mice were suppressed by concentrations of anti-mu several hundred-fold lower than required for adult spleen cells; the adult pattern of susceptibility to suppression was acquired by 1 week of age. No significant differences in susceptibility to anti-mu were found in comparisons of adult spleen, lymph node, bone marrow, and Peyer's patch lymphocytes.


The merrit alloantigenic system of human B lymphocytes: evidence for thirteen possible factors including one six-member segregant series. An analysis of the typing results of a 70-member chronic lymphatic leukemia B cell panel revealed evidence for 13 possible groups of the Merrit alloantigenic system. Six of these appeared possibly allelic and may represent a segregant series. The CLL panel was also fully typed for HLA and some degree of linkage dysequilibrium between Merrit and HLA seemed apparent from the data. Merrit antibodies can be absorbed out with selected surface membrane immunoglobulin (SMIG)-positive normal lymphocytes and less so or not at all with E rosette-forming T cells or Fc-positive SMIG-negative lymphocytes.


Bernard-Soulier syndrome: a new platelet glycoprotein abnormality. Its relationship with platelet adhesion to subendothelium and with the factor VIII von Willebrand protein. A decreased platelet adhesion to rabbit aorta subendothelium (Baumgartner technique) is confirmed in the Bernard Soulier (giant platelet) syndrome. Electron microscope techniques using a purified antibody against Factor VIII/von Willebrand protein, revealed an apparently normal presence of the Factor VIII/von Willebrand protein on the Bernard Soulier platelets. Electrophoretic characterization of the major protein and glycoprotein components of the Bernard Soulier platelets following sodium dodecyl sulfate solubilization indicated a relatively normal protein content but suggested a reduced content of the 155,000 molecular weight major platelet glycoprotein. This was confirmed by a reduced release of high molecular weight acidic glycopeptides following incubation of washed Bernard Soulier platelets with trypsin. It is proposed that this abnormality may be related to the previously reported reduced sialic acid content and the reduced electrophoretic mobility of the Bernard Soulier platelets and that a glycoprotein reduced or abnormal in the Bernard Soulier platelets is necessary for the normal adhesion of platelets to subendothelium.


Simplified alpha1-antitrypsin phenotyping by immunofixation of acid-starch gels. Immunofixation of acid-starch gels represents a simplified method for alpha1-antitrypsin phenotyping. This technique reduces the amount of work and time involved in phenotyping by eliminating the need for crossed-immunoelectrophoresis. Its major purpose is to directly enhance the staining quality of the minor bands, especially the Z bands on the acid-starch gel. It is hoped that this modified method will allow more wide-spread use of antitrypsin phenotyping, especially in the routine clinical chemistry laboratory.


Embryological evidence for a possible polyphyletic origin of the recent amphibians. The markedly different mode of mesoderm formation in anuran and urodelan amphibians (which is related to the early double-layered nature of the anuran blastula wall in contrast to its single-layered nature in the urodeles), but particularly the fundamentally different place and mode of origin of the primordial germ cells in the two groups of amphibians, strongly pleads in favour of a very ancient bifurcation in the phylogenetic history of the two groups, even suggesting a polyphyletic origin from different ancestral fishes.


The initiation of diving apnoea in the frog, Rana pipiens. 1. Diving apnoea in Rana pipiens was initiated by submerging the external nares. As the water level was raised above the frog, both buccal and lung pressure increased by an amount corresponding to the water head. During submergence the external nares remained closed, although the apnoeic period was punctuated by ventilation movements which moved gas between the lungs and buccal cavity. 2. Bilateral section of the ophthalmic nerves did not alter the normal pattern of ventilation in air, although it often resulted in the intake of water into the buccal cavity on submergence. Introduction of water into the buccal cavity, either naturally as in denervates or by injection through a catheter in intact frogs, triggered sustained electromyographical activity in some respiratory muscles. 3. Electroneurograms recorded from the cut peripheral end of an ophthalmic nerve showed that receptors in the external narial region were stimulated by movement of a water meniscus across them. Activity could also be recorded in the ophthalmic nerve in response to water flow past the submerged nares. Punctate stimulation of the narial region confirmed that these receptors were mechanosensitive. 4. Bilateral electrical stimulation of the central ends of cut ophthalmic nerves in lightly anaesthetized frogs caused apnoea with a latency of less than 200 ms. The external nares remained closed throughout the period of stimulation although buccal pressure events, resembling underwater ventilation movements, occurred when stimulation was prolonged.


Separation of helper T cells from suppressor T cells expressing different Ly components. I. Polyclonal activation: suppressor and helper activities are inherent properties of distinct T-cell subclasses. Concanavalin A, a nonspecific polyclonal activator of T lymphocytes, activates Lyl and Ly23 subclasses to the same degree. After activation, the Ly23 subclass, but not the Lyl subclass, has the following properties: (a) Suppression of the antibody response to sheep erythrocytes (SRBC) in vitro. (b) Production of a soluble factor that suppresses the anti-SRBC response in vitro. (c) Suppression of the generation of cell-mediated cytotoxicity to H-2 target cells in vitro. Con A-activated cells of the Lyl subclass, but not the Ly23 subclass, express helper function in the anti-SRBC response in vitro. Because the intact Con A-stimulated T-cell population contains both cell types, these cells do not exert detectable helper effects in an anti-SRBC system in vitro, because the helper effect of Lyl cells is masked by the suppressor effect of the Ly23 cells. Each function is revealed by eliminating one or the other population with the relevant Ly antiserum. The resting T-cell population, before activation by Con A, also contains already programmed Lyl and Ly23 cells with similar helper and suppressor potentials, respectively. This is revealed by experiments with Ly subclasses which have been separated from the resting T-cell population and then stimulated by Con A. Thus helper and suppressor functions, as expressed in these systems, are manifestations of separate T-cell-differentiative pathways and do not depend upon stimulation of the cells by antigen.


Separation of helper T cells from suppressor T cells expressing different Ly components. II. Activation by antigen: after immunization, antigen-specific suppressor and helper activities are mediated by distinct T-cell subclasses. Cells of the Lyl subclass generate helper activity in both primary and secondary responses to sheep erythrocytes (SRBC). In contrast, after priming with SRBC, cells of the Ly-2+ subclasses, in particular Ly23 cells, have suppressive activity. The degree of Ly23-mediated suppression is directly proportional to the amount of antigen (SRBC) used for priming. Suppression by Ly23 cells is specific, because Ly23 cells from SRBC-primed animals do not suppress the response to horse erythrocytes, and vice versa. Thus, both cytotoxic and specific suppressor functions are mediated by T cells of a subclass, provisionally designated TCS, which can be distinguished from helper T cells (TH), by their Ly phenotypes. It remains to be determined whether killing and suppression are functionally interrelated properties of a single Ly23 subclass, or whether the Ly23 population comprises two subclasses whose surface phenotypes are not yet distinguishable by immunogenetic criteria.


Ontogeny of B-lymphocyte function. II. Ability of endotoxin to increase the heterogeneity of affinity of the immune response of B lymphocytes from fetal mice. The ontogeny of the functional capacity of B lymphocytes to generate a heterogeneous response to a haptenic determinant was studied by cell transfer techniques in LAF1 mice. Fetal liver, as a source of B lymphocytes, was transferred into adult, syngeneic, irradiated animals. All recipients received excess adult thymus cells so that T-cell activity did not limit the response and were immunized with DNP-BGG. The heterogeneity of avidity of their anti-DNP PFC response was assayed by hapten inhibition of plaque formation. Animals reconstituted with B lymphocytes from fetal donors produced a response that is highly restricted with respect to heterogeneity of affinity. Transfer studies using multiple fetal donors or mixtures of adult and neonatal cells for reconstitution suggest that the restriction in heterogeneity is not the consequence of suppressor T-lymphocyte activity. With animals reconstituted with B cells from day 16 or older fetal donors, injection of LPS together with antigen converted the response to a heterogeneous "adult-type" response. With animals reconstituted with B lymphocytes from day 14 fetal liver DxSO4, but not LPS, could convert the response to a highly heterogeneous one. Animals reconstituted with day 14 or 16 fetal liver as source of B lymphocytes were capable of producing a heterogeneous secondary response despite the fact that their primary response was of restricted heterogeneity. This implies the selection of high affinity B-memory cells, in the absence of high affinity PFC during the primary response with fetal B lymphocytes. Animals reconstituted with day 14 or 16 fetal liver produce only direct PFC, while animals reconstituted with day 18 fetal liver produce both direct and indirect PFC. Three differentiation events have therefore been defined in the functional development of B lymphocytes: (a) between day 14 and day 16 of fetal life they acquire responsiveness to LPS; (B) BETWEEN DAY 16 AND 18 OF FETAL DEVELOPMENT THEY ACQUire the capacity to produce indirect PFC; (C) between day 7 and 10 after birth they acquire the capacity to give a heterogeneous response after normal immunization. In addition, it was shown that LAF1 mice already have all of the information required to produce an "adult-type" heterogeneous anti-DNP response at day 14 of fetal life.


Lysozyme synthesis by established human and murine histiocytic lymphoma cell lines. A human cell line established in culture from a histiocytic lymphoma patient synthesizes and secretes the monocyte-granulocyte specific enzyme lysozyme. 18 other human cell lines with characteristics of T-lymphocyte, B-lymphocyte, Burkitt's lymphoma, non-Burkitt's lymphoma, myeloma, and bone marrow epithelial cells were not associated with lysozyme. Among murine cell lines, lysozyme was produced by (a) three histiocytic lymphoma or macrophage lines, which mediate antibody-dependent phagocytosis and cytolysis; (b) myelomonocytic leukemia line which also secretes myeloid colony-stimulating factor; and (c) a spontaneous lymphoma and an Abelson leukemia virus-induced lymphoma. Lysozyme-negative lines include another Abelson lymphoma, myelomas, T lymphomas, and mastocytoma.


Cell-mediated immunity: delayed-type hypersensitivity and cytotoxic responses are mediated by different T-cell subclasses. Cell-mediated immunity includes both the generation of cytotoxic cells and initiation of delayed-type hypersensitivity (DTH). The resting T-cell population, before stimulation by antigen, already contains cells of the Lyl subclass that are programmed to initiate DTH (and helper function) but not cytotoxic responses, as well as Ly23 cells which can generate killer activity (and suppressive function) but not DTH. The central implication of these findings is that the broad division between humoral and cell-mediated immune responses does not precisely correspond to the division of labor among T-cell subclasses. The relative contribution of DTH-competent Lyl cells and cytotoxic Ly23 cells to the classical homograft response remains to be determined.


Cellular basis of neonatal induction of in vitro tolerance. The LD and SD antigens of the major histocompatibility complex subserve differential roles in the induction of the proliferative phase in mixed lymphocyte culture and in the cytotoxic reaction seen in cell-mediated lympholysis. The present study suggests that they also behave differently in the neonatal induction of tolerance. SD antigens appear to induce tolerance in the cytotoxic T lymphocytes very effectively, whereas LD antigens (or the cytotoxic targets coded by genes in the I and/or S regions) are relatively ineffective in this regard. LD antigens presented neonatally are effective at inducing tolerance in the proliferating helper cells.


Fetal antigens on the surface of human lymphoid cells. Human B-lymphoblastoid cells established in long-term culture from healthy adults carry surface components that are normally found in human fetal tissues at about 10 wk of age. These antigens are strongly expressed on neoplastic B lymphocytes but not on thymocytes or a cultured T-cell line. They are carried by a small subpopulation of normal adult peripheral blood lymphocytes as well.


An estimation of the frequency of precursor cells which generate cytotoxic lymphocytes. The cell-mediated immune response has been generated in vitro with a polyacrylamide culture system which allows the segregation of foci (clones?) of cytotoxic lymphocytes. Using the method of limiting dilutions, the frequency of precursor cells in CBA spleen cells able to generate a cytotoxic response against DBA mastocytoma is estimated at 1 per 1,700 cells.


Density and distribution of tetrodotoxin receptors in normal and detubulated frog sartorius muscle. Tetrodotoxin (TTX) binding was measured in muscles which were either in normal condition or which had been "detubulated" by glycerol-induced osmotic shock. In both cases the binding of TTX was found to saturate at high TTX concentrations. Maximum binding in normal fibers was 35 pmol/g wet weight, and that figure was reduced to 16 pmol/g after glycerol treatment. The dissociation constant for binding to the surface membrane was 3 nM, which is the range of values obtained by electrophysiological measurements of the effect of TTX on the maximum rate of rise of the action potential. The results suggest that the dissociation constant in the transverse tubules may be higher than that in the surface. Control experiments indicate that the effects of glycerol treatment are limited to the accessibility of the receptors to the toxin and result in no alteration of the affinity of the binding site. TTX receptors in the transverse tubules may be recovered after glycerol treatment by homogenization of the fibers. The measurements suggest that the density of sodium channels in surface membrane is about 175/muM2 and that in the transverse tubular membrane is 41-52/mum2.


Normal rate of cerebrospinal fluid formation five years after bilateral choroid plexectomy. Case report. A ventricular perfusion technique was used to determine the rate of cerebrospinal fluid (CSF) formation in a 5-year-old child who had undergone bilateral choroid plexectomy for communicating hydrocephalus during infancy. At the time of the study, the patient had a failed ventriculoperitoneal shunt and was suffering from progressive ventriculomegaly. The calculated rate of CSF formation, 0.35 ml/min +/- 0.02 standard deviation, was within normal limits.


The effects of serotonergic and antiserotonergic drugs on the flexor reflex of spinal rat: a proposed model to evaluate the action on the central serotonin receptor. The effects of serotonergic and antiserotonergic drugs on the hind limb flexor reflex (measured as a contraction of musculus tibialis anterior or as a flexion of the paw) in the spinal rat was studied. All serotonergic drugs used (L-5-hydroxytryptophan, L-tryptophan, LSD, fenfluramine, p-chloroamphetamine) stimulate the flexor reflex. Serotonin receptor blockers (cyproheptadine, WA-335, methergoline), which given alone are inactive, inhibit the stimulation induced by serotonergic drugs but do not influence that one caused by noradrenergic agents (amphetamine, clonidine). Both types of stimulation (serotonergic and noradrenergic) are antagonized by noradrenaline receptor blockers (phenoxybenzamine, clozapine). The antiserotonergic action of serotonin antagonists used was confirmed on the basis of the antagonism to the head twitches induced by L-5-hydroxytryptophan. The flexor reflex in the spinal rat may be a good model to evaluate the effect on the central (spinal cord) serotonin receptor and to differantiat this effect from that on the noradrenaline system.


A new concept in the control of acute gingival hemorrhage. The topical application of epsilon-aminocaproic acid may be life-saving therapy for control of acute gingival hemorrhage in patients who are debilitated because of systemic diseases. When low doses were used, there have been no reports of adverse reactions.


Continuous bilateral electrophrenic pacing in an infant with total diaphragmatic paralysis. An infant with bilateral diaphragmatic paralysis, etiology undetermined, is presented in whom bilateral intrathoracic phrenic nerve pacemakers were inserted. He was maintained with continuous bilateral electrophrenic pacing for 142 days. A review of the literature by the authors has not revealed any previously reported cases.


Safety aspects of mandibular lingual surgery. Mandibular lingual surgery is commonly used in periodontal practice. There are inherent risks in such surgery which are illustrated by a case report described here. This was a case in which an emergency tracheotomy was needed after a minor procedure which involved lingual tissues. The potential problems of any form of lingual surgery are discussed. Various other surgical procedures in this region are examined in order to establish some surgical principles which may be of use to periodontists.


Effects of ethanol on the permeability of frog skin. Ethanol (3%) decreases the potential difference and short-circuit current across the isolated frog skin in chloride Ringer's solution. Unidirectional fluxes of Na and Cl indicate that the drop in short-circuit current is due to an inhibition of the sodium influx. However, ethanol had no effect on the electrical parameters or sodium fluxes, when the frog skin was bathed in chloride-free solutions on both sides or the outside alone. The ethanol response is anion-dependent. In addition, chloride-free media in the inside bathing solution reduced the short-circuit current, indicating a sodium transport pathway which is dependent on chloride and confirming previous data in the literature. Other anions such as sulfate and nitrate could not substitute for chloride. The vasopressin-induced natriferic response and the ethanol effect were found to work independently of each other and different pathways of action are suggested for these agents. The intracellular sodium content of the isolated frog skin epithelium increased and potassium decreased in the presence of the Na-K adenosine triphosphatase inhibitor, ouabain, whereas ethanol or amiloride had no effect. The oxygen consumption of the isolated frog skin was unaffected by up to 10% ethanol. A general metabolic action is probably thus not mediating the response. Urea, in iso-osmotic concentrations to the ethanol, did not mimic its effect. Tritiated water fluxes (in the absence of an osmotic gradient) were reduced by 30% in the presence of 3% ethanol. It is suggested that ethanol may impede the flow of water across frog skin by a physicochemical interaction with membrane pores and the water molecules. The permeability coefficient (Ktrans) for ethanol was found to be 10 times smaller than the Ktrans for water.


Components of human split ejaculates. II. Enzymes and proteinase inhibitors. Lysozyme, alpha-amylase, neutral proteinase and plasminogen activator were most concentrated in the initial portion of the ejaculate that consists mostly of Cowper's gland and prostate gland fluids as well as spermatozoa. The concentration of the high molecular weight proteinase inhibitors, alpha1-antitrypsin and alpha1X-antichymotrypsin, was essentially unaltered throughout the ejaculate fractions, although their absolute amounts showed an increase towards the final fraction. By contrast, the total inhibitory activity towards pancreatic trypsin was highest both in concentration and amount in the last fraction, thus indicating that the seminal vesicles are its primary source. Plasminogen, prothrombin, Factor XIII, and the proteinase inhibitors antithrombin III, alpha2-macroglobulin, inter-alpha-trypsin inhibitor and C1S-inactivator could not be detected immunochemically in whole ejaculates, and indicates the dissimilarity between the coagulation/liquefaction processes of semen and blood.


'Durate variant with clinical signs' has alpha1 -antitrypsin genotype ZZ. A patient with neonatal jaundice and cirrhosis who was previously reported homozygous for the Durate variant of galactose-1-phosphate uridyl transferase has the ZZ genotype for alpha1-antitrypsin. A sister of the patient, also with ZZ genotype, is less severly affected with liver disease and is a heterozygote for the Durate variant. Since a number of patients with ZZ genotype of alpha1-antitrypsin have been previously reported to have liver disease, the latter genotype is the more probable explanation for the patients' clinical state. A question is raised, however, whether the Duarte variant may be specifically associated with the development of liver disease in ZZ individuals.


Effects of small organic molecules on the cholinergic receptors of the motor endplate. This paper deals with the effects on the endplate receptors of the electro-osmotic and bath application of compounds which, although lacking pharmacological affinity, mimic the actions caused at much lower concentrations by conventional cholinergic drugs. Small cyclic molecules containing carbonyl, ether, ester, amide, and carboxyl anhydride groups were tested. Lactones, carboxyl anhydrides, cyclic amides, and ketones depolarized the endplate membrane and, in addition, most of them potentiated the depolarizing action of acetylcholine. On the contrary, cyclic ethers behaved as neuromuscular blocking agents. Dioxane, which contains two ether oxygens, caused a marked prolongation of the time course of the potentials elicited by the iontophoretic application of acetylcholine pulses. In some instances the effects of adding the compounds to the bath were different from those observed following local electro-osmotic application.


Location in bacteriophage lamdba DNA of cleavage sites of the site-specific endonuclease from Bacillus amyloliquefaciens H. The sites in Escherichia coli bacteriophage lambda DNA cleaved by the site-specific endonuclease isolated from Bacillus amyloliquefaciens H (BamI) are found to be at 0.114, 0.466, 0.580, 0.713, and 0.861 lambda units. The sites were located by analysis of the products of digestion of lambda DNA and lambda-ara transducing phage DNA, and verified by double digestion with BamI and EcoRI.


Prophage induction and inactivation by UV light. Analysis of the induction curves for UV light-irradiated Haemophilus influenzae lysogens and the distribution of pyrimidine dimers in a repair-deficient lysogen suggests that one dimer per prophage-size segment of the host bacterial chromosome is necessary as a preinduction event. The close correlations obtained prompted a renewed consideration of the possibility that direct prophage induction occurs when one dimer is stabilized within the prophage genome. The host excision-repair system apparently functions to reduce the probability of "stabilizing" within the prophage those dimers that are necessary for induction and inactivation. The presence of the inducible defective prophage in strain Rd depresses the inducibility of prophage HP1c1.


Immune reactivity in renal cancer: a sequential study. We evaluated 18 patients with adenocarcinoma of the kidney immunologically in a linear fashion. Delayed cutaneous hypersensitivity was significantly altered in patients with metastatic disease. In vitro studies have shown a progressive decline in the absolute number of circulating lymphocytes and the T cell subpopulations that parallels the advance of the maligant process. Evaluation of lymphocyte function by the response to mitogens appears to be of little significance in a sequential study.


[Analysis of some parameters for determining myocardial contractility before and after aortocoronary shunting]. The condition of hemodynamics and the contractile function of the myocardium are considered before and after aorto-coronary shunting performed in patients with critically severe affection of the coronary arteries and the heart muscle. Of great prognostic importance is the final diastolic volume of the ventricles, its relation to the stroke ejection, as well as the ejection fraction. The greater the final diastolic volume of the ventricle and the smaller the stroke ejection, the higher is the risk of applying operative treatment to the patient.


Organ-specificity and diagnostic value of cell-mediated immunity against a liver-specific membrane protein: studies in hepatic and non-hepatic diseases. In chronic active hepatitis (CAH, n=58) 70% of the HBsAg negative and 48% of the HBsAg positive cases showed a CMI against human liver specific proteins (HLPI). Using HBsAg as antigen only 12% of the HBsAg negative and 24% of the HBsAg positive cases gave a CMI response. On the basis of HBsAg and autoantibodies in the serum CAH patients could be divided into 4 subgroups. A close correlation between CMI against HLPI, sex, ANA and HL-A-8 could be detected. In a follow-up study of patients with acute virus B hepatitis (n=62) CMI against HBsAg was detected in 60% of the cases in the acute phase of the disease but in 15% only 3-6 months after the onset of the illness (n=40). In patients who developed a chronic HBsAg carrier status 3 of 5 cases remained persistently positive with HLPI as antigen in the migration inhibition test. - In non-hepatic diseases in which immunological abnormalities may be present (malignant diseases n=46, diabetes mellitus n=27, active tuberculosis, n=18 and untreated systemic lupus erythematodes, n=5) only 26% of patients with malignant diseases showed a migration inhibition with HLPI. - Using different antigens such as human liver specific proteins (HLP), rabbit liver specific proteins (RLP), brucella suis antigen and tuberculin it was possible to demonstrate the validity of the two-step migration inhibition test to detect CMI. The results with different antigens in hepatic and non-hepatic diseases demonstrated that cell-mediated immunity of HLPI is an organ specific immune reaction which is associated with acute and chronic active liver diseases as a time limited or long-lasting phenomenon. Positive reactions in some tumor patients suggest that different mechanisms may elicit an autoimmune reaction against liver antigens.


[Use of specific anti-T-lymphocyte globulin (sATG) for the diagnosis of lymphoproliferative diseases (author's transl)]. Difficulties in the production of specific antisera against T-lymphocytes could be overcome by a stepwise absorption and purification procedure of anti-human thymocyte serum. Specific anti-T lymphocyte globulin (sATG) reacted with thymocytes, thymus-derived lymphocytes and a lymphoblastoid cell line of T-cell type whereas no activity was found against lymphoblastoid cell lines of B-cell type. Five chronic and three acute lymphatic leukemias were characterized using sATG in the cytotoxic test, electron microscopy, complement fixation test and quantitative immunoautoradiography, and compared with lymphocyte populations of normal individuals. Three chronic lymphatic leukemias with low numbers of spontaneous rosettes and high percentages of membrane-Ig-positive lymphocytes showed only few T-cell-antigen-positive lymphocytes and were therefore classified as B-cell leukemias. The cells of two chronic lymphatic leukemias with high numbers of spontaneous rosettes carried T-cell-antigen. The T-cell-antigen concentration, however, was lower than that of normal peripheral blood T-lymphocytes. The T-cell nature of two acute lymphatic leukemias with high numbers of spontaneous rosettes was confirmed by a positive reaction of the cells with sATG. In one case of acute lymphatic leukemia most leukemic cells carried T-cell-antigen although these cells did not form spontaneous rosettes. In the first two cases the T-cell-antigen concentration on the cell surface exceeded that of normal blood-T-lymphocytes, in the latter case it was slightly below that. The advantages of the characterization of leukemias with sATG in comparison with the spontaneous rosette formation and the relevance for prognosis are discussed.


Valvulotomy of valves in the saphenous vein graft before coronary artery bypass. Occasionally, repeat angiograms disclose turbulence, with or without clot formation, at the valve sites in saphenous veins used as coronary artery bypass grafts. A simple valvulotomy may be used to split these valves to allow a more even flow and to avoid the valve recess where air may be trapped or thrombus formed. A potential cause for perioperative infarction or graft failure may thus be avoided.


Upper gastrointestinal haemorrhage-a review of 100 cases. The case records of the last 100 consecutive patients with upper gastrointestinal tract bleeding admitted to the adult section of the Mater Misericordiae Hospital have been analysed with attention to presentation, investigations, aetiology, management and mortality. The period of these admissions was almost three years-from May, 1971, to June, 1974.


Arbovirus infection in a Murray Valley community. I. Prevalence of antibodies, December, 1974. A survey of antibodies to Group A and Group B arboviruses (also designated as alphaviruses and flaviviruses respectively) was performed in residents of a Murray Valley town eight months after an outbreak of arbovirus illnesses in other parts of the region. The sampling pattern was based upon a random selection of households. The prevalence of Group A antibodies rose steadily with relation to age. The prevalence of Group B antibodies was much lower in all age groups, and did not show such a consistent rise with age. No Group B antibodies were found in younger residents. This suggests either an unexpectedly rapid decay of the particular type of antibody in this age group, or regional variations in the prevalence of arbovirus which are not currently understood. Some individuals appear to be unusually exposed to arbovirus infection.


Topographical analysis of yeast ribosomal DNA by cleavage with restriction endonucleases. Yeast ribosomal DNA (rDNA) was digested with the restriction enzymes Hind III, Hind II and a mixture of Hind II and Hind III. The cleavage products were analyzed by electrophoresis on 1.5% agarose gels. Several distinct bands could be observed, which are derived from the redundant ribosomal transcription units. They are superimposed on a rather broad smear of background DNA, representing the heterogenous 'spacer' sequences. From the restriction maps, together with data obtained by partial digestion, a physical map for the ribosomal transcription unit in yeast could be constructed.


E rosettes in lymphoproliferative diseases. Results are given of investigations of E rosettes in patients with chronic lymphadenosis and malignant lymphoma. The quantity of E rosettes was expressed both as per cents and in absolute numbers calculated from the total amount of lymphocytes in peripheral blood. In the normal controls the percentage of E rosettes was 58% on average while in patients with chronic lymphadenosis it was found to have decreased significantly down to a mean of 6%. However, in absolute values the number of rosette forming lymphocytes was normal and even higher. In the group with malignant lymphoma the percentage decrease was less striking than in those with chronic lymphadenosis, however, the absolute number of rosette forming lymphocytes was always found to be lowered because of evident to considerable lymphopenia. A dynamic study of the observed changes might contribute to a knowledge of the pathophysiology of lymphoproliferative diseases.


The effect of apomorphine and ergocriptine on the release of MSH by the pars intermedia of Rana pipiens. In an attempt to further characterize the nature of the innervation controlling melanophore-stimulating hormone (MSH) release from the pars intermedia (PI), pellets containing either apomorphine or ergocriptine were implanted in contact with the PI of the frog, and the effect on the animal's coloration was observed. Frogs with apomorphine implants exhibited skin pallor for several hours and those with ergocriptine implants were pallid for a few days. Hypophysectomized (HX) frogs which had been darkened with MSH regained their pallor no sooner when given an injection of apomorphine than with the vehicle. However, intact dark frogs lightened when injected with apomorphine. Frogs darkened by autotransplantation of the PI to the dorsal lymph sac were lightened by an injection of apomorphine. Frogs lightened by an ergocriptine implant on the PI and then darkened by MSH did not lighten sooner than HX frogs given an injection of MSH. Ergocriptine-containing pellets implanted in the thigh took longer to produce the same effect than those implanted in contact with the PI. Since apomorphine is known to stimulate dopamine receptors in the central nervous system, and there is some evidence that ergocriptine does also, the above evidence supports the hypothesis that dopamine receptors are present in the PI.


Disseminated intravascular coagulation and fibrinolysis as a cause of postextraction hemorrhage. Report of a case. Presented is a case in which severe, prolonged hemorrhage, subsequent to simple dental extractions in an 85-year-old Negro man, was due to disseminated intravascular coagulation, with secondary fibrinolysis resulting from metastatic adenocarcinoma of the prostate gland. The difficulty and delay in diagnosis, selection of proper therapeutic regimen, and complications encountered during treatment are discussed.


Voltage clamp analysis in isolated cardiac fibres as performed with two different perfusion chambres for double sucrose gap. Voltage clamp experiments were performed on isolated frog atrial trabeculae disposed in 2 different perfusion chambres for double sucrose gap. In one of the perfusion chambres, a "liquid partition system" (L.P.S.) was used; in the other, a vaseline sealing (V.P.S.) method was used to separate the various fluid compartments. From the linear electrical properties, the elements of an apparent equivalent electrical circuit were calculated. The apparent nodal capacity was significantly larger in fibres disposed in the "liquid partition system" than those disposed in the "vaseline partition system". The apparent "error factor" resulting from the presence of the series resistance was larger in the L.P.S. than in the V.P.S. The apparent "error factor" becomes relatively large when the membrane conductance increases, such as during the flow of the inward current. A rapid desactivation of the peak inward current was found on bringing the command potential back to the resting potential in the V.P.S. This was not found in the L.P.S., indicating better voltage control with the V.P.S. than with the L.P.S. Transmembrane microelectrode recordings during voltage clamp experiment in the V.P.S. indicated satisfactory voltage control during the flow of the peak inward current. Adequate voltage control is lost when notches or irregularities appear on the current traces.


[Early functional disorders in 3 children exhibiting an obstructive syndrome and an alpha 1 antitrypsin deficiency]. The respiratory function is studied in 3 children, two of which 4 years in age, exhibiting a respiratory obstructive syndrome and an alpha1-antitrypsin deficiency. The study of the respiratory function is global and regional. The disorders noted include a ventilatory asynchronism, a distension, regional anomalies of ventilation and perfusion. The ventilatory asynchronism appears to be an early disorder in the course of alpha1-antitrypsin deficiency. These results have to be related with the development of the elastic fibers during the child's growth.


Separation of inner and outer membranes of Rhodopseudomonas spheroides. The separation of inner and outer membrane of Rhodopseudomonas spheroides has been achieved by means of sucrose density gradient (20%, 40%, 60%, w/w) centrifugation. The upper fraction of the gradient, with a specific density 1.181 (g/cm3), is high in cytochrome and succinate dehydrogenase activities, low in lipopolysaccharides and it is designated the inner membrane fraction. The bottom fraction of the gradient, with a specific density 1.240, is high in lipopolysaccharide and contains neither cytochrome nor succinate dehydrogenase activities. This fraction is the cell wall or outer membrane fraction. The intermediate band on the gradient is an unseparated fraction of inner and outer membrane fragments. This fraction has a specific denisty of 1.211 and represents less than 3% of total crude envelope. Thin sections of the vesicles of the inner membrane fraction and those of outer membrane provide morphological evidence for the identity of the individual membrane fractions. At least 22 protein bands are resolved by employing sodium dodecyl sulfate slab gel electrophoresis. Six bands are present only in the inner membrane and two bands are found exclusively in the outer membrane. Most of the remaining polypeptides are present in greater amounts in the inner membrane relative to the outer membrane fractions.


[Biosynthesis of 5- methoxy-N-acetyltryptamine (melatonin) in enterochromaffin cells]. Chromatographic analysis of the alkaline extract of the gastrointestinal mucosa of rabbits rich in enterochromaffin cells displayed the presence of melatonin (5-methoxy-N-acetyltryptamine) and its precursors (5-oxytryptophane, tryptamine, serotonine, mexamine) in it. Detection of precursors indicated that not a passive accumulation, but an active synthesis of melatonin occurred in the enterochromaffin cells. Epiphysis was not the only place of melatonin synthesis. Functional condition of enterochromaffin cells was largely determined by the extent of physiological and pathological reactions of the organism.


Antibody-producing cells: virus-induced alteration of response to antigen. Spleen antibody-forming cells of mice yield a 3- to 10-fold increase in their response to sheep erythrocyte antigen if they are acutely infected by lactate dehydrogenase-elevating virus. This early stimulation is replaced by a long-term inhibition of the antibody-forming cells as the viremia goes into its persisting chronic stage. These contrasting immunological phenomena are examined as contributing factors responsible for the enhancement by this virus of asparaginase (EC 3.5.1.1; L-asparagine amidolydrolase) therapy against leukemia in mice, and for the alteration of the susceptibility of mice to various neoplastic processes.


[Electrophoretic mobility of human lymphocytes purified by nylon columns and spontaneous rosettes]. Cell electrophoresis enables the separation of the lymphocytes in normal human blood into two principal groups, as a function of their migration speed in relation to 1 mum.sec -1..v-1.cm. In 42 healthy adults, 19,9 % of the lymphocytes have a slower migration, and 80,1 % a faster migration than the reference speed. Two known methods are used for the selection of the lymphocytic populations : spontaneous rosetting with sheep red blood cells, a property of the T lymphocytes, and the adherence to nylon wool columns, which is dominant in the case of B lymphocytes. The cells which do not form spontaneous rosettes, and the cells adhering to nylon wool columns have above all a slow migration. On the contrary, cells which do not adhere to nylon columns have a fast migration. These arguments are in favour of the T nature of the rapid migrating lymphocytes, and of the B nature of the slow migrating lymphocytes.


[Xeroradiography in rheumatology].

[Xeroradiography in rheumatology]. Xeroradiography is a new and simple radiodiagnostic technique that makes it possible to analyse on one positive film not only the bone tissue but also the neighbouring structures (ligaments, muscles, fatty tissue, the vessels, and the skin). On the basis of about 100 examinations, the authors indicate the value and the limitations of the technique in rheumatological practice. Because of the technical aspects of the technique, the dorsolumbar spine and the facial massif are poorly visualized. The swamping of the contrast also limits its use for cases of demineralization (osteoporosis). On the other hand, the information provided by the technique in cases of Paget's disease, articular or para-articular calcification, pathological conditions of the tendons (rupture, calcification), and bone tumours appears to be of great value. The early diagnosis of cases of inflammatory rheumatism and the surveillance of Silastic prostheses used in surgery on rheumatic hands are also facilitated.


[Osteoid osteoma. 5 cases].

[Osteoid osteoma. 5 cases]. With reference to five cases of osteoid osteoma, including two para-articular cases with articular reaction, two cases with neurological manifestations, and one case with a rare costal lesion, the authors demonstrate the difficulty of diagnosis in this disease, which is, in principle, well known. In some cases the observations extended over seven years. The authors also review the literature on this subject.


[New research on synovial enzymology].

[New research on synovial enzymology]. After reviewing previous work on the subjects, the authors show that the synovial fluid in subjects with inflammatory rheumatism contained higher levels of lysozyme and of beta-glucuronidase comparable with those of the acid phosphatases and of lacto-dehydrogenase that they were interested in previously.


[Rheumatic manifestations in 80 cases of Crohn's disease]. Out of a series of 80 patients suffering from Crohn's disease, 31 presented rheumatic manifestations. In 16 subjects this took the form of synovitis closely dependent on the enteritic evolution, which developed after the alimentary symptoms, and which worsened as they did and sometimes regressed as they did following medical or surgical treatment. In combination with erythema nodosum, aphtosis, and conjunctivitis, synovitis appears to be the expression of an immune response to the enteritic lesion. Three cases of chronic polyarthitis and 6 cases of asymptomatic sacro-ileitis were also observed, and 6 cases of spondylarthritis of a minor radiological type were observed that evolved independently of the Crohn's disease. Typing according to the HLA system using 26 antigens was carried out in 44 subjects; no difference in phenotype frequency was found between a control group (blood donors) and the group of subjects with Crohn's disease alone; however, the antigen W 17 was found significantly more frequently in those subjects with peripheral arthritis and the antigen W 27 was found more frequently in those with spondylarthritis. These findings suggest, although it is not certain, the existence of genetic susceptibility to rheumatic manifestations in certain sites in patients with Crohn's disease.


[Pigmented villonodular synovitits of the hip: ultrastructure and aspects on scanning electron microscopy]. The authors studied one case of pigmented, villonodular synovitis (PVNS) of the hip by means of optical microscopy and by transmission and scanning electron microscopy. Scanning electron microscopy showed that the surface of the PVNS is completely different from that of rheumatic synovitis, in particular that of rheumatoid synovitis. The composition and the cellular morphology of the outer layer of the PVNS appear, however, to be similar to those of normal synovial membrane of arthrosic synovial membrane. Clumps of red corpuscles, enclosed in a fibrin network, were visible on the surface of the PVNS in a way that the authors has never seen previously in the 19 other human synovial membranes, normal and pathological, they had studied in this way. The totality of the microscopic findings confirms the importance of the role played by the intra-articular and intra-synovial haemorrhages, and by the macrophage reactions that follow, in the development of the lesions that characterize PVNS.


Evidence that carrier-specific B cells play a role in cellular collaboration. Evidence is presented showing that carrier-specific B cells play a significant role in inducing an anti-hapten antibody response to the thymus-dependent antigen, dinitrophenylated monomeric flagellin (DNP-MON), both in vivo and in vitro. Inactivation of carrier-specific B cells by (125I) polymeric flagellin (POL) suicide decreased the anti-DNP IgM response by 40%-50% and reduced the anti-DNP IgG response by 85%-90%. Several lines of evidence established that the suicide procedure eliminated carrier-specific B cells rather than carrier-specific T (helper) cells.


The effect of anti-immunoglobulin antibodies of Fc rosettes. This study demonstrated that Fc rosettes detected by antibody-labeled bovine erythrocytes can be inhibited by immune complexes of rabbit IgG (b4 allotype) and anti-b4 IgG, thus showing that these complexes effectively compete for the receptor for erythrocyte-antibody rosettes. Incubation of mouse lymphocytes with goat F(ab')2 anti-mouse IgG did not inhibit Fc rosettes. When immunoglobulin on murine and human B lymphocytes was redistributed in a polar cap on the membrane by interaction with anti-immunoglobulin antibodies, the number of Fc rosettes after 'capping' was unchanged; most of the indicator erythrocytes, however, were distributed in a cap as well. This erythrocyte cap was superimposed on the immunoglobulin cap.


The role of B-cell memory in secondary IgG and IgM responses. Mice were primed with the hapten 3-nitro-4-hydroxy-5 iodophenacetic acid (NIP) conjugated to chicken globulin (cg) and were boosted 2, 6, or 12 months later with CG conjugates of the related haptens 3,5-diiodo-4-hydroxyphenacetic acid (DIP) or 3-nitro-4-hydroxphenacetic acid (NP). Accelerated secondary responses were demonstrated both in the 7S and 19S class. Fine-specificities of secondary-response antibodies were studied by the hapten inhibition method of haptenated bacteriophage inactivation. 7S antibodies were found to have the fine-specificity of anti-NIP antibodies regardless of whether DIP or NP was the booster hapten ('original antigenic sin'). 19S antibodies had the fine-specificity of anti-DIP when DIP was the booster hapten. NP as the booster hapten resulted in 19S antibodies whose fine-specificity was intermediate between anti-NIP and anti-NP. A strong B-cell memory could thus be demonstrated in the 7S antibody response and a weak B-cell memory in the 19S antibody response.


Distribution of cells binding erythrocyte-antibody (EA) complexes in human lymphoid populations. Cells bearing Fc receptors were studied using a rosette technique with chicken erythrocytes coated with rabbit antibodies (EA). By comparison with other markers and by selective depletion of T and B cells it was demonstrated that EA rosettes were formed by about 10% of the monocytes, 7% of the T lymphocytes, and 45% of the B cells. In addition, 26% of EA-rosette-forming cells (EA-RFC) in the peripheral blood carried on other markers than C3 receptors. Marked differences were found in the percentages of EA-RFC in cell suspensions from various lymphoid organs: peripheral blood, 15.2%; spleen, 26.6%; bone marrow, 30.9%; lymph nodes, 2.8%; thymus, 0.3%; and tonsils, 0.5%. It was concluded that Fc receptors were present on subsets of different lymphocyte subpopulations; their presence could not be regarded as a specific marker of one of these subpopulations.


Antibody-dependent cell cytoxicity (ADCC). Characterization of 'killer' cells in human lymphoid organs. The effector cell(s) in human antibody-dependent cell cytotoxicity (ADCC), with antibody-coated chickens erythrocytes as targets, was studied by comparison of cell suspensions from various lymphoid organs and by means of various cell fractionation methods. Effector cells (K) were found mostly in peripheral blood, spleen, and bone marrow but not in tonsils, lymph nodes, and thymus. Effector cells bear Fc receptors and can form EA rosettes with the antibody-coated target cells. About 1,5% peripheral blood lymphocytes can form 'high-avidity' EA rosettes with targets coated at low antiserum concentration. Most of the effector cells belong to this small subset, as shown by experiments of selective depletion. Removal of most monocytes, T cells, or B cells from, or addition of T-cell-specific antiserum to, the effector cell suspensions did not affect ADCC. Effector cells in this model of ADCC therefore lack the conventional B- or T-cell markers but at least some of them are likely to bear C3 receptors.


Does nonspecific T-lymphocyte stimulation of B lymphocytes occur during reversal reaction in borderline leprosy? Serum immunoglobulins G, A, and M were estimated in 14 patients with border-line cases of leprosy at commencement of treatment and subsequently when they developed 'reversal reaction'. There was a significant increase in all immunoglobulin levels during the reaction, with a subsequent fall; the postreaction values for IgG and IgA were below the base-line figures. Additonal investigations in six patients indicated that the rise was a nonspecific one, not brought about by an increase in antimycobacterial antibodies. It seems likely that the rise in immunoglobulins during reaction is due to nonspecific T-lymphocyte stimulation of B lymphocytes.


Endoscopic removal of gastric and duodenal polyps. As endoscopic surgery comes into its own, the applications of this modality are increasing. This is well illustrated by the use of endoscopic polypectomy in the colon, and more recently by endoscopic polypectomy in the stomach and duodenum. We can anticipate increased applicability of these technics as the experience widens and the margins of safety increase. The present series of six polypectomies from the stomach and duodenum confirms the applicability of endoscopic polypectomy for this portion of the gastrointestinal tract, attests to its safety, and indicates that these procedures can contribute materially to the care of patients with gastric or duodenal polyps. The real and potential problems are discussed.


Coagulation studies in leptospirosis.

Coagulation studies in leptospirosis. Coagulation and serum F.D.P. studies in 10 leptospirosis patients (5, L. bataviae, 2, L. autumnalis, 2, L. australis, and 1, L. akiyami A.)showed prolongation of prothrombin time, partial thromboplastin time and thrombin clotting time, slight low platelet count, depletion of factor V and slightly high serum F.D.P. in only four cases. Four cases had bleeding and one case died because of severe haemorrhage but with only slight changes in coagulation factors and slightly high serum F.D.P. These findings suggested that the damage of capillary endothelium was more pronounced than the coagulopathy which may be due to liver cell damage.


A bleeding syndrome in infants: acquired prothrombin complex deficiency of unknown aetiology. A total of 240 cases of a bleeding syndrome in infants due to prothrombin complex deficiency of unknown aetiology were reviewed. The majority of patients were breast fed, aged 1-2 months and the syndrome was more prevalent in males. Clinical manifestations consisted of bleeding, pallor and mild hepatomegaly in the majority of cases. Mild fever, diarrhoea, jaundice, and upper respiratory tract infection were associated in a few patients. Acute onset, short course and a high rate of intracranial bleeding (65%), particularly subdural and subarachnoid, were observed. The haemostatic defects appeared to be a marked reduction in factor II, VII, IX, and X. Complete blood counts remained relatively normal, with the exception of some changes (anaemia, leukocytosis), in response to the acute bleeding. Liver chemistry was normal or slightly impaired. No specific pathological changes were noted at autopsy, there were mild changes of liver cells, such as rare focal necrosis of liver cells, the proliferation of Kupffer cells, extramedullary haemopoeisis and mild cholestasis. Clinical improvement and correction of hemostatic defects were noted after vitamin K therapy alone or with fresh blood transfusion. Mortality rates were high in infants with intracranial bleeding (40-55%), while the overall mortality rate was 25%. The pathogenesis and the possibility of prevention of the syndrome were discussed.


Rupture of the left kidney following renosplenic shunt. Kidney rupture following division of the left renal vein during portal decompression is presented. A review of the renal venous anatomy is described and a possible explanation of this ususual occurrence is offered.


The late results of terminal esophagoproximal gastrectomy (TEPG) with intensive devascularization and splenectomy for bleeding esophageal varices in cirrhosis. The operative procedure of terminal esophagoproximal gastrectomy (TEPG) with extensive esophagogastric devascularization and splenectomy is described in detail and its rationale is discussed. Late results of this operation performed therapeutically on 64 patients with posthepatitic cirrhosis revealed that (1) operative mortality rate (death within a month) was 11%, mostly being contributed by poor-risk patients; (2) over-all survival was 65% at 5 years and 59% at 10 years; (3) gastrointestinal bleeding recurred in nine patients, five of whom had major and four minor hemorrhages; (4) portasystemic encephalopathy was seen frequently after shunts had never occurred, and the hepatic encephalopathy was only a sequel of major gastrointestinal bleeding or a manifestation of terminal-stage liver insufficiency; and (5) ascites was not a major clinical problem after this operation; mild esophagitis was an occasional complication.


[The tourniquet-syndrome--a severe complication after embolectomy of saddle embolism (author's transl)]. The symptoms of the tourniquet syndrome (hypotonia, tachycardia, postischemic edema, hypercaliemia, metabolic acidosis, myoglobinuria, renal insufficiency) could be observed after embolectomy of saddle embolism in 37 patients. 19 patients died postoperatively; heart insufficiency was demonstrated by autopsy in 14 patients. The pathophysiology of heart failure in tourniquet syndrome was studied in dog experiments after unilateral and bilateral hind limb ischemia. The development of shock turned out to be more severe after bilateral ischemia--comparable to saddle embolism--than after unilateral ischemia. The course of heart failure after recirculation could be referred to hypercaliemia, hypermagnesiemia, metabolic acidosis and hemoconcentration. The prophylaxis of the tourniquet syndrome can be practised by knowing the pathogenesis.


[Early postoperative diagnostic management by arterial-reobstruction after vascular operations using the ultrasonic flowmeter (author's transl)]. The Doppler ultrasonic method can confirm the success of the operation in the early post-operative phase after reconstructive vascular operations as an essential non-invasive technique or clear a doubtful improvement of the blood flow. Thus an arteriography can be indicated in good time. On the basis of reconstructive cases the usefulness and the advantageous application of the Doppler ultrasonic flowmeter in the operating theater is demonstrated.


[Paraplegia following acute infrarenal aortic occlusion (author's transl)]. The clinical syndrome of paraplegia following acute occlusion of the infrarenal aorta may be caused by either ischemic spinal cord damage or ischemia of the cauda equina and sacral nerve roots and ganglia. The neurologic manifestations are similar and therefore specific anatomic diagnosis is difficult. From October 1972 to February 1975 a total of 31 patients with infrarenal aortic occlusion were treated at the Medizinische Hochschule in Hannover. In nine cases the occlusion up to the renal arteries was acute. Three of these patients presented beside acute ischemic manifestations on both legs and the lower abdominal wall, neurologic symptoms of paraplegia. The anatomic and hemodynamic aspects of ischemic spinal cord damage and those of ischemic lesions of the cauda equina and peripheral nerves are discussed. There appear to be three main mechanisms responsible for vascular paraplegia following acute infrarenal occlusion of the aorta: 1. it may be caused by thrombotic occlusion of a major radicular artery which arises below the level of occlusion. 2. it may be produced by thrombosis of a lumbar collateral acting as major supply to the cord when arteriosclerotic narrowing of the major radicular artery is present. Especially in states of severe hypotension critical interference of blood supply to the spinal cord will result. 3. Paraplegia by ischemia of the cauda and peripheral nervous tissue may also follow prolonged interruption of circulation to this area supplying spinal vessels from low lumbar and sacral arteries.