Tonic influence of the efferent vestibular system on the spontaneous afferent activity from semicircular canals in the frog (Rana esculenta L.). In the frog we have recorded the spontaneous activity of single afferent fibres of the ampullary nerves of the left horizontal semicircular canal (HC) and vertical anterior canal (VAC) in isolated head preparations. The recordings have been made in 4 experimental situations: intact preparations; preparations whose brain was destroyed; preparations whose contralateral vestibular nerve had been cut between Scarpa's ganglion and the brain; preparations where either an ampullary nerve (that of the HC or of the VAC) or the utricular nerve had been cut contralaterally. From our results, it appears that the efferent vestibular system has a tonic influence on the afferent activity from HC and VAC; the influence of the part of the efferent vestibular system (EVS) activity depending on contralateral vestibular inputs is inhibitory, while the influence of the part of the EVS activity depending on ipsilateral vestibular inputs might be facilitatory.


Levels of alpha1-antitrypsin in the spring and autumn seasons. In a group of 84 pairs of 11-year-old children of both sexes, the level of the alpha1-antitrypsin (alpha1-AT) were ascertained in the autumn and spring. Although the mean levels of alpha1-AT in the two seasons hardly differed, the highly significant seasonal changes in the distribution curves of alpha1-AT values were noted in boys, whereas the levels showed higher stability in girls.


The loss of biological activity of 5-hydroxytryptamine creatinine sulphate. 5-Hydroxytryptamine creatinine sulphate loses its biological activity when maintained at room temperature. The loss of 5-HT activity (in stimulating sodium transport across frog skin) is greater than the loss of creatinine sulphate activity (inhibition of sodium transport).


Depletion of synaptic vesicles at the frog (Rana pipiens) neuromuscular junctions by tetraphenylboron. At the frog neuromuscular junction tetraphenylboron produced a decrease in miniature endplate potential amplitude, accompanied by a decrease in the number but not the size of synaptic vesicles.


Does an initial phasic response exist in the receptor potential of taste cells? The depolarizing receptor potentials to 0.5 M NaCl recorded from frog taste cells did not exhibit any phasic response, even when the rectangular waveform of stimulus onset was employed. The quickest depolarizations recorded reached the peak in 50 msec. On the other hand, the gustatory neural response showed initial overshoot of the impulse discharge even when 0.5 M NaCl was delivered at the slower rate of 0.06 ml/sec. It is concluded that the initial neural response may be associated with the rate of rise of the receptor potential before its plateau level is reached.


[Oriented rat movement in a rotating system]. In experiments with rotating of rats, their avoidance responce was studied: rats moved into the central portion. The 7 degrees or 15 degrees pitch of the central portion's walls caused significant moving of rats towards a square peripheral portion. When experimenting in darkness, the moving--towards--center response was restored, while under illumination the preference of the peripheral portion was again revealed in the rats obtained are are regarded as confirming the hypothesis of taht the moving--to center response during rotation is due to discrepancy between the vestibular and the visual systems of coordinates in peripheral portions of centrifuge.


[Inhibitory influence on the heart of antidromal stimulation of afferent fibers of the vagus nerve]. In partially or completely deparasympathized frogs, inhibition was not potentiated by the specific inhibitor of "true" cholinesterase acetylcholinesterase: the 1,5-bis(4-tri-methylammoniumphenylpeptan-3-on-diiodide). The localization of cardiac "true cholinesterase" being determined only in the region of the postsynaptic membrane of post-ganglionic neurons, this enzyme lacking on their extrasynaptic surface and in the myocardium, it was concluded that excitatory effect of antidromic afferent activation on the intracardiac neurons was due to action of acetylcholine (released from the nerve endings) on the extrasynaptic region of neurons' membrane. These regions acquire cholino-receptive features after preganglionic parasympathetic degeneration.


[Relationship between the strength of myocardial fiber contraction of frog heart ventricle and processes of intracellular energy transport]. At 10--15 mM of creatine in the perfusate the force of the myocardium contraction was increased. At concentrations over 40 mM, creatine causes a decrease of contractile force which can be significantly increased after washing-out of the excess of creatine. Prolonged perfusion (over 10 hrs) with Ringer's solution leads to a decrease in intracellular creatine concentration and to a parallel decrease of contractile force. Under these conditions the intracellular concentration of creatine phosphate is significantly decreased with small changes in ATP content. The addition of creatine to perfusate leads to the complete restoration of contractile force and creatine phosphate content in heart strips' cells. These results indicate that the contractile force of heart strips depends upon high energy phosphate content (mainly creatine-phosphate content) in the cells and are consistent with the idea of the existence of the creatinephosphate pathway of energy transport in heart cells.


Electrokinetic properties of splenic lymphocytes from the low-lipopolysaccharide responder C3H/Hej mice. Spleen cells from C3H/Hej mice (H-2k) respond poorly to the B-cell mitogen lipopolysaccharide in vitro as compared to the related strains C3H/Tif (H-2k) and CBA/Orl (H-2k). The electrokinetic properties of splenic lymphocytes from these 3 strains were investigated in parallel, in order to both quantitate low-mobility B cell and high-mobility T cell populations and measure their mean electrophoretic mobilities. C3H/Hej mice were found to possess the same proportion (55%) of LM cells as C3H/Tif and CBA/Orl mice. Therefore, the low LPS-responsiveness of C3H/Hej is not due to a numerical deficiency in B cells. Whereas the mean EPM of HM cells was identical in the 3 strains, that of LM cells was slightly (6%) but significantly (Student's t test, P less than 0.01) lower in C3H/Hej than in the high LPS-responder controls. This suggests that the membrane-structure required for activating interaction with LPS might contribute to B-cell electronegative surface-charge.


Changes in agglutination of cells with H-2 antibodies or lectins and in stimulation of lymphocytes as induced by certain polymers (chondroitin sulphate/PVP/PHA/Con A/capping of H-2 antigens). The effect of certain polymers on the course of immune interaction has been well known although its mechanism at the cellular level remains obscure. We thus affected cells in vitro with two of such substances, CS and PVP, and followed their behaviour in various assays. It was shown that in this way the agglutinability of mouse erythrocytes with H-2 antibodies or of mouse lymphocytes with threshold doses of Con A could be enhanced, antibody-induced capping of H-2 antigens on lymphocytes accelerated and PHA-induced stimulation of mouse lymphocytes inhibited. The possible mechanisms of these changes putatively mediated through the effect on the cell membrane or cell coat are discussed.


Aspects of lymphocyte function in Behçet's disease. Aspects of lymphocyte function were investigated, in vitro, in nine patients with active Behçet's disease. The rosette formation technique revealed a slight decrease in number of T lymphocytes and more so of B lymphocytes. In spite of their low number, the transformation of the T lymphocytes in response to mitogens did not show a significant difference from that of normal controls. This may indicate a hyperreactivity, and in two instances a spontaneous blastogenesis prior to addition of mitogens, was also noted. The immunoglobulins and complement levels in serum were within normal limits.


Hemangiomas of the cecum. Colonoscopic diagnosis and therapy. Five cases of hemangiomas of the cecum and gastrointestinal bleeding are presented. All were demonstrated by colonscopy after more conventional diagnostic methods failed. The hemangiomas were bright red, flat lesions clearly seen through the colonscope. They were successfully treated by electrocoagulation. All patients in this series had some type of associated cardiac or vascular disease. These patients may represent a syndrome of gastrointestinal blood loss of obscure cause, hemangiomas of the cecum, and cardiovascular disease.


[Evaluation of the agglutinating action of lectins on Trichomonas vaginalis by means of different methods]. Trichomonas vaginalis (strain FC) are specifically agglutinated by Con A and PHA-P but not by WGA. The techniques used gave basically the same results however the agglutination in plates has the advantage of a good sensitivity and a greater simplicity of execution.


[Capacity of activation of so-called deficient factors in hereditary blood coagulation disorders]. By factor-specific exchange-experiments it was proved that the lack of factor VIII activity in hemophilia A is not caused by an incapacity of activation or even lack of the factor, but by a defect of the pertaining activator resp. activator system. The same goes for factor IX in hemophilia B as well as factors VII, IX and X in family defect of the prothrombin-complex. In contrast, inactivity in the von Willebrand-Jünrgens-disease is obviously caused by a genuine lack of factor VIII.


[Hemorrhagic enteropathy].

[Hemorrhagic enteropathy]. Intestinal infarction in the absence of organic vascular occlusion received increasing attention in recent years. The clinical picture is discussed based on results in 9 cases, an attempt to suggest a possible pathophysiological mechanism is made. Prophylactic digitalisation especially in the elderly patient in the absence of severe heart failure and in cases with already low mesenteric perfusion may lead to a further vasoconstriction and to hemorrhagic enteropathy. Therapeutic possibilities are discussed.


[Clinical importance of Meckel's diverticulum]. Between 1966 und 1975 42 children and 46 adults were operated on Meckel's diverticulum. The diverticulum is explained as one of the possible disturbances during regression of ductus omphaloentericus. The appendicitis-like symptomatology correlates to the involvement of gastric and colonic mucosa as well as heterotopic exo- and endocrine tissue of the pancreas in the wall of the diverticulum. In 34% complications were due to inflammation, perforation, bleeding, intussusception, volvulus, gut-strangulation with ileus, ulcer, neoplasia or lesion by foreign bodies. 4 out of 88 patients died. Preoperative diagnostic fails in 75%, therefore in every case the distal gut should be inspected and every diverticulum should be resected.


[Diagnostic and therapeutic measures in acute catastrophic bleeding esophageal varices]. Principles of management of bleeding esophageal varices are 1. fluid therapy of bleeding shock, 2. prevention of hepatic coma, 3. emergency endoscopy, 4. balloon tubes (Senkstaken-Blakemore, Linton-Nachlas), and 5. with some restriction, selective infusion of vasopressin into the a. mesenterica superior. If these procedures fail, sclerosing of esophageal varices stops bleeding in more than 90% of the cases. Bleeding from varices of the gastric fundus may be stopped by gastro-esophageal disconnection (Pettinari-Hassab). Both procedures have with 15% and 25% respectively, the lowest mortality. Patients for surgical shunt are carefully selected within the interval after bleeding. Shunts are the distal splenal-renal and the mesenteric-caval anastomosis with dacron prothesis (H-shunt). The shunt is the favorable therapy for prehepatic block in patients older than 14 to 16 years. The endoscopic sclerosing of esophageal varices and the gastro-esophageal disconnection are chosen in younger patients or when shunt procedures are not possible. The posthepatic block is treated successfully by conservative means. In most cases, surgical therapy is contraindicated because of poor prognosis. When conservative measures fail, in few cases emergency endoscopic sclerosing of esophageal varices or latero-lateral porto-caval anastomosis can be tried.


Elastases from human and canine granulocytes, I. Some proteolytic and esterolytic properties. The lysosome-like granules of human and canine granulocytes contain an enzyme with elastinolytic activity. The enzymatic behaviour of these elastases was further characterized using the protein substrates elastin-orcein and azocasein and the synthetic substrates tert.-butyloxycarbonyl-alanine p-nitrophenylester (Boc-Ala-ONp) and 3-carboxypropionyl-L-alanyl-L-alanyl-L-alanine p-nitroanilide (Suc-Ala3-NHNp) in photometric assays. The affinities of the granulocyte elastases and of porcine pancreatic elastase to these substrates are very similar, e.g. human granulocyte elastase: KM (Boc-Ala-ONp) = 0.35mM, KM (Suc-Ala3-NHNp) = 1.25mM, porcine pancreatic elastase: KM (Boc-Ala-ONp) = 0.3mM, KM (Suc-Ala3-NHNp) - 1.15mM. The most convenient substrate for the assay of human and dog granulocyte elastases and for kinetic measurements with these enzymes is Suc-Ala3-NHNp. Using this substrate, the dissociation constant of the complex of human granulocyte elastase with human alpha1-antitrypsin could be determined (Ki = 3.5 x 10(-10)M).


The function of T cells carrying receptors for complexes of Ig and antigen. Thymocytes exposed briefly in vitro to a variety of particulate substances (such as mycobacteria, erythrocytes of allogeneic cells) or to substances known to act in vivo as adjuvants (LPS or poly A:U), generate supernates which are able to induce cytophilic Ig in normal mouse serum in the presence of a foreign protein (antigen). This cytophilic Ig is taken up by 20-25% of splenic T cells. Hydrocortisone resistant thymocytes show the same property, while bone marrow cells are inactive. This activity is similar to that reported previously as being present in the 4S fraction of mouse serum, collected 6 hours after injection of complete Freund's adjuvant. It is proposed that this factor is responsible for the formation of complexes of Ig and antigen which have been detected in the serum 6 hours after immunization. Thymocytes collected 6 hours after priming in vivo with SRBC (when a subpopulation among them carries easily demonstrable surface Ig) are able to amplify markedly the antibody response particularly the 7S. It is postulated that the factor by generating the cytophilic Ig (complexes?) which is taken up by T cells, sets up a mechanism which markedly amplifies their helper cell function.


Competitive inhibition of passive sensitization of mouse mast cells by IgE. A bioassay for mouse and rat IgE. Possibility of inhibition of an efficient in vitro IgE-sensitization system was studied. The sentization of mouse peritoneal mast cells with an anti-ovalbumin IgE-rich fraction of serum, as tested by ovalbumin-induced degranulation, was inhibited by previous incubation with antisera of another or of no specificity. Fractionation and other experiments showed that the inhibiting activity correlated with IgE content. IgGl did not seem to have an effect. Sensitization was also inhibited by rat myeloma IgE, 50 ng giving a 50 per cent inhibition. Plots of the logarithms of rat and mouse IgE concentration vs their inhibitory effect on sensitization gave two parallel linear curves, indicating that mouse and rat IgE compete for the same receptor sites. It was thus possible to use this system as a sensitive bioassay for both mouse and rat IgE levels and, by comparing inhibition by mouse IgE to that by a known rat IgE standard, to obtain not only relative data but absolute mouse IgE levels. This, and also a better discrimination of IgE doses, was the major advantage of this bioassay in relation to the equally sensitive anti-IgE degranulation tests.


Study of human T and B lymphocytes with heterologous antisera. I Preparation, specificity and properties of antisera. Heterologous antisera have been raised in the horse and rabbit against human lymphocytes. Appropriate absorptions on either B or T cells were performed to make antisera specific for human T (anti-HTLA) or B (serum 789) lymphocytes respectively. In addition serum 789 was found to react with circulating monocytes. The percentages of T and B cells detected by anti-HTLA and 789 sera in the different lymphoid organs averaged respectively: 78-7 per cent and 14-7 per cent in peripheral blood, 91-4 per cent and 4-0 per cent in thymus, 73-0 per cent and 14-5 per cent in lymph nodes, 53-6 per cent and 30-0 per cent in spleen, 47-1 per cent and 47-6 per cent in tonsils and 17-5 per cent and 13-5 per cent in bone marrow. Anti-HTLA serum appeared to supress E-rosette formation but did not affect binding of C3-coated erythrocytes. Serum 789 did not prevent E-rosette formation and reduced the number of EAC rosettes by 50 per cent. Anti-HTLA serum was found able to suti-lymphocyte serum, and PWM in the presence of complement; it was found highly mitogenic by itself. Serum 789 decreased the proliferative response to phytomitogens in about the proportion of cells killed by the antiserum. These results indicate that the presence of T cells is necessary for the mitogen-induced proliferation to occur, and that B lymphocytes are induced to proliferate in the presence of T cells and phytomitogens. Anti-HTLA serum was found not to inhibit K-cell activity of lymphocytes against antibody-coated chicken erythrocytes. These antisera appear very useful tools for the study of the role of human B and T lymphocytes involved in in vitro immune reactions.


Activation and suppression of T cells in termination of immunological tolerance. Immunological tolerance was induced in adult C3H mice by five injections of ultra-centrifuged, soluble HSA at weekly intervals. The tolerant state was readily terminated by the challenge immunization with slightly sulphanilated HSA administered as Freund's complete adjuvant emulsion. In this case, it was shown that T cells specific for HSA were activated which could serve as helpers for the production of anti-hapten antibodies. The tolerance termination and activation of T cells specific for HSA was not observed in the profound tolerant state, induced by nine injections of ultra-centrifuged, soluble HSA at weekly intervals, where suppressor T cells were detected. Implications of these results were discussed in reference to the development of autoimmunity.


Rapid adsorption of a foetal calf serum component by mammalian cells in culture. A potential source of artifacts in studies of antisera to cell-specific antigens. Injection of CBA mice the either mitogen-stimulated (LPS or con A) CBA lymphocytes which had been cultured for 3 days in the presence of foetal calf serum (FCS) led to the production of antisera which reacted strongly with virtually all types of mammalian cells, including human, whether normal or malignant, provided they had been cultured in FCS-containing media. Reactivity was detected by sensitive immunological assays such as complement-dependent cytoxocity using rabbit complement (but not using guinea-pig complement), or EA-rosette inhibition of Fc receptor-bearing cells. The antisera did not react with fresh normal lymphoid cells or ascites tumour cells; however, these same cell populations became fully susceptible to the cytotoxic effects of the antisera after as little as 4 h incubation at 37 degrees in the presence of FCS. Cells incubated without FCS or with FCS at 0 degrees were not affected. The antisera reacted with FCS to form a single band on Ouchterlony double-diffusion plates. On immunoelectrophoresis the reactive antigen appeared to migrate in the alpha-globulin region of serum proteins. These observations suggest that FCS may be a source of potentially serious misinterpretations in immunological studies of cell-associated antigens using antisera produced by the injection of cells grown in FCS-containing cultures. Examples of artifcats arising from the use of FCS in certain systems, e.g. the preparation of alloantisera using cultured tumour cells vs fresh non-cultured lymphoid cells, are described.


An IgG thymolytic autoantibody in rats which has specificity for a subpopulation of T cells. A cytotoxic anti-thymocyte IgG auto-antibody is present in Lewis rats which, in the presence of autologous complement, destroys (in vitro) 12-28 per cent of isologous or autologous thymocytes, a smaller number of lymph node cells and splenocytes, but not bone marrow or circulating lymphocytes. The labile cells in the thymus represent a finite subpopulation which is autologous antithymocyte antibody (ATS) sensitive and steroid resistant. The presence of the autoantibody is randomly distributed in outbred animals whereas in inbred Lewis rats, a strain in which the induction of some autoimmune reactions is under genetic control, the antibody is always present. In this strain, the susceptible T cells and the quantity of circulating autoantibody is significantly depressed during the productive phase of a T-cell mediated disease (adjuvant polyarthritis) and returns to normal after the disease becomes stabilized. There is a direct relationship between the amount of susceptible cells in the thymus and the amount of antibody in circulation, suggesting that the antibody could serve as a marker for a specific subpopulation of thymocytes which may have a regulatory influence on T-cell reactivity.


Experimental endogenous endophthalmitis caused by Haemophilus influenzae type b. Experimental endogenous endophthalmitis was produced in infant rats by either intranasal or intraperitoneal inoculation with Haemophilus influenzae type b and 5 days of age. The ocular disease occurred in about 50% of bacteremic animals who survived to age 12 days and probably represents metastatic bacterial infection secondary to hematogenous seeding. The lesion was a highly destructive suppurative endophthalmitis that ultimately progressed to panophthalmitis and was followed by organization of the exudate and phthisis bulbi.


Participation of complement in the nonimmune host defense against experimental Haemophilus influenzae type b septicemia and meningitis. This study was undertaken to determine whether the terminal complement components (C3-9) are involved in the nonimmune host defense against Haemophilus influenzae type b septicemia and meningitis. Using cobra venom factor, infant rats were depleted of C3 and C5. After intranasal challenge with H. influenzae type b, the complement-depleted rats developed a greater incidence and magnitude of bacteremia and a higher mortality rate. In contrast to the effects on bacteremia, complement depletion did not directly influence either the occurrence of meningitis or bacterial multiplication within the cerebrospinal fluid. These experiments provide evidence that the complement system may be an important mechanism of natural immunity to H. influenzae type b.


A comparison of three health status indicators. Interest in health status indicators has produced measures of widely varying applicability. We were interested in the use of such indices to establish mean recovery curves for groups of similar patients undergoing acute hospital treatment. A longitudinal study relating resource usage to these recovery curves had been intended but there were difficulties in finding suitable indicators. Firstly, two published indicators relying on patient interviews were tested for consistency. Poor correlations were found among those scorers unfamiliar with the patients and it seems unlikely that these indicators could be used in a routine system. Different parts of the indices presented difficulties to the different professions involved in scoring, and a multidisciplinary approach may be needed in assessing full health. The indicators tested included no assessment of prognosis. Those parts of the indices which had produced significant correlations were retained in subsequent work and were supplemented by further measures designed to overcome the earlier difficulties. A new trial of this indicator was undertaken where staff, familiar with the patients, scored data recorded by the Problem Oriented Medical Record system. This produced improved correlations but some problems remain.


The Tokelau Island migrant study: prevalence of various conditions before migration. The Tokelau Island Migrant Study has shown no important differences between those who subsequently left their home islands to migrate to New Zealand and those who remained, in key anthropometric and biochemical variables already reported. This comparison is now extended to various common diseases and conditions, and again no major difference emerges. The Tokelauans are compared with other Polynesians and shown to have less diabetes, hypertension, effort pain, chronic bronchitis and varicose veins than New Zealand Maoris, while resembling some Cook Island groups. Changes in prevalences of some conditions following migration are postulated.


Regeneration and dose-response characteristics of irradiated mouse dorsal epidermal cells. A microcolony technique is described for measuring epidermal cell survival 3 days after whole-body X-irradiation. This assay provides a cell D0 value of 233 +/- 11 rad and a zero-dose extrapolate of 1-23 x 10(4) cells/cm2 for mice irradiated in oxygen 20 hours after hair plucking. The microcolony cellularity had an apparent doubling-time of 25 hours which may be an upper limit at least for some clones. The clones appeared to fragment continually and form new daughter clones, suggesting that few would form macroscopic nodules. Many of the clones were also apparently associated with hair follicles.


Effect of 2 MHz ultrasound on DNA, RNA and protein synthesis in Pisum sativum root meristem cells. The amounts of DNA, RNA and protein synthesis were determined in Pisum sativum root meristem cells at various times after a 1 min exposure to 1 MHz ultrasound at a power density of 30 W/cm2. Immediate depressions in all three macromolecular syntheses occurred after sonication, followed by an apparent recovery several hours later. These events appear to correlate in time with the subsequent reduction and recovery in mitotic index in Pisum sativum root meristem cells exposed to 2 MHz ultrasound.


Biological relevance of gamma-ray-induced alkali-labile sites in single-stranded Dna in aqueous solutions. Gamma-irradiation in single-stranded phiX174 DNA in aqueous solution in the presence of oxygen produces at least two types of alkali-labile site. One is lethal and gives rise to single-strand breaks by treatment with alkali. The other is non-lethal, but is converted to lethal damage by alkali. The effect of alkali is dependent on temperature. This dependence is different for both types of alkali-labile site.


X-irradiation of equine peripheral blood lymphocytes stimulated with phytohaemagglutinin in vitro. Small lymphocytes were isolated from the peripheral blood of horses and incubated at 37 degrees C in Eagle's medium supplemented with 20 per cent foetal calf serum. The addition of phytohaemagglutinin (PHA) to the cultures resulted in: increased RNA and protein synthesis; the enlargement of the small lymphocyte into a lymphoblast-like cell; the initiation of DNA synthesis, and cell division. When survival was measured 24 hours after X-irradiation by means of phase-contrast microscopy, the lymphoblast-like cell was much more radio-resistant (D0 = 250 rad) than the small lymphocyte (D0 = 20 rad). This increase in radioresistance, however, was not observed until 12-24 hours after PHA treatment. To investigate which of the changes occurring during the transformation of the small lymphocyte was responsible for the increased resistance to irradiation, the percentage of cells surviving irradiation was compared with the percentage of cells incorporating significant amounts of 3HTdR, 3H-UR, or 3H-leucine at the time of irradiation. For this comparison, a dose of 100 rad was used because 100 rad killed essentially all of the small lymphocytes, but less than 35 percent of the cells which had become radioresistant from the PHA treatment. The results indicated that the increase in radioresistance was not associated with DNA synthesis, but instead correlated with the increase in RNA and protein synthesis which the cells had attained at the time of irradiation.


The effect of nitroimidazole and nitroxyl radiosensitizers on the post-irradiation synthesis of DNA. The modification of DNA damage by three radiosensitizing drugs, present during gamma-irradiation of hypoxic Chinese hamster cells, was investigated. Both 2-methyl-5-nitroimidazole-1-ethanol (metronidazole) and 1-(2-nitro-1-imidazole)-3-methoxy-2-propranol (Ro-07-0582) were found to cause large increases in the yield of DNA single-strand breaks (SSB); triacetoneamine-N-oxyl (TAN) was found to have only a small effect on SSB production. The three drugs tested did not inhibit the rejoining of SSB. A pulse label and chase procedure was used to examine post-irradiation DNA synthesis. TAN present during irradiation under hypoxia was found to cause interruptions in subsequent DNA synthesis. Metronidazole and Ro-07-0582 had no effect on post-irradiation DNA synthesis. In addition, the effects of pre- and post-irradiation exposure to TAN were investigated, since these treatments have shown increased cell-killing in survival studies. TAN pre- and post-treatments were found to have no significant effect on subsequent DNA synthesis.


Viable hybrids between lethally X-irradiated hamster cells and unirradiated mouse cells. Sendaï-virus-induced fusion between heavily X-irradiated hamster cells, BHK21 or RS2-3 (BHK21 cells transformed by Rous Sarcoma Virus), and unirradiated mouse cells, A9 or c11D, give rise to hybrids. These hybrids possess mouse and hamster surface antigens. However, RS2-3 x mouse hybrids do not form heterokaryons with chick-embryo fibroblasts producing infectious Rous sarcoma virus.


R.b.e. and o.e.r. of extended-Bragg-peak helium ions: survival and development of rat embryos. Rats were exposed under aerobic or hypoxic conditions to 200-1200 rads of 60Co gamma-rays or extended-Bragg-peak helium ions on the eighth day of gestation. Uterine contents were examined on the twentieth day of gestation. At the 50 per cent embryonic survival level, helium ion r.b.e. was 1(.0) (aerobic) and 1(.2) (hypoxic). Maximum attainable gamma-ray and helium-ion o.e.r.s. were 2(.2) and 1(.7) respectively, indicating an oxygen-effect gain (o.e.g.) of 1(.2). At the 10 per cent survival level helium ion r.b.e. was 1(.1) (aerobic) and 1(.4) (hypoxic). Gamma-ray and helium-ion 0.e.r.s. were 2(.0) and 1(.5) respectively, indicating a helium ion o.e.g. of 1(.3). These data demonstrate that the small fraction of high-LET radiation present in this helium ion beam has a neglible effect on the aerobic r.b.e., but lowers the effective o.e.r. of the beam approximately 25 per cent relative to that of gamma-rays. Helium ions were significantly more effective than gamma-rays in killing embryos under hypoxic conditions, in producing congenital abnormalities under aerobic conditions, and in stunting foetal growth under both conditions.


Effects of electrons and neutrons on the synthesis of RNA in resistant and sensitive strains of the slime-mould Dictyostelium discoideum and the modifying effect of caffeine. RNA synthesis was investigated after irradiation in resistant and sensitive lines of the slime-mould Dictyostelium discoideum. When 3H adenine was used as a precursor to RNA, incorporation increased after irradiation in the resistant WT line but not in the sensitive line (gammas-13). The extent of RNA synthesis after irradiation was correlated with the shoulder width on the survival curve of the resistant line. When this was reduced by irradiating with neutrons, or treatment with caffeine RNA synthesis was also reduced. No preferential synthesis of one RNA species occurred; there was increased labelling in all RNA species after irradiation. Sucrose gradient analysis of ribosomal RNA extracted from irradiated cells and free of messenger RNA revealed no apparent difference in composition from that extracted from unirradiated cells. Increased RNA synthesis after irradiation may form part of the recovery processin the resistant cells.


Analysis of the time relationship for the interaction of X-ray induced primary breaks in the formation of dicentric chromosomes. In split-dose experiments, the time relationship for the interaction of primary breaks in the formation of dicentric chromosomes was analysed. Human peripheral lymphocytes were irradiated with a dose of 340 R of 220 kV X-rays split into two equal fractions separated by intervals up to 360 min. Assuming an exponential decline of the time-dependent quadratic component of the dose realtion of dicentrics a theoretical formula was deduced. Fitting this formula to the empirical data a mean time of 110 min could be calculated in which primary breaks induced by both dose-fractions can interact to form dicentric chromosomes.


Abscopal effect of irradiation on haemopoietic stem cells of shielded bone marrow--role of migration. Immediately after partial-body irradiation, the number of colony-forming units (CFU) of the protected marrow decreases. This decrease seems to be dose-dependent and cannot be completely explained by migration for the following reasons: there is no evidence of CFU entry to irradiated territories and the number of circulating CFU does not increase. A more plausible explanation for the phenomena is an accelerated differentiation during the first 15 min after irradiation. The migration of CFU to the spleen is observed between 15 min and 3 hours after irradiation, after which it ceases. The absence of migration 3 hours after irradiation may be due to one of the two possibilities: (1) a depletion of the mobile pool of CFU if one admits that such a pool is responsible for migration in the organism; (2) a depletion of a temporarily increased stimulus due to irradiation.


Vasoformative non-osteogenic (angio) sarcomas of bone-marrow stroma due to strontium-90. In a series of experiments, mainly CBA/H, but also C2H/H, mice aged 3 months were injected intraperitoneally with solutions of 90Sr Cl2, the dose per mouse varying from 7 to 20 muCi, and compared with similar mice treated with 226Ra or 239Pu, discussed elsewhere. In male mice, the commonest tumour resulting at each dose of 90Sr was non-osteogenic (angio) sarcoma, a tumour not seen after 226Ra. In females, this tumour occurred far less frequently than osteosarcoma. In CBA mice of both sexes converted to radiation chimaeras (which are sterile) and similarly treated with 90Sr, the only skeletal tumours were osteosarcomas. When only half the body of CBA mice was X-irradiated with 1000 rad and the mice given 90Sr, non-osteogenic sarcoma occurred predominantly in those mice X-irradiated in the cephalic half. The results suggest that intact testes may provide co-factors for this type of neoplasm, whereas others have shown that oestrogens facilitate murine osteosarcoma. The non-osteogenic osteosarcomas arise from damaged stromal elements in bone-marrow of selected bones. The risk to this component of bone-marrow, as well as to haematopoietic tissue, should be considered in radiation protection.


Inhibition of prostaglandin-mediated ocular inflammatory responses by 4-biphenylacetic acid. 4-Biphenylacetic acid (BPAA),a prostaglandin-synthesis inhibitor, was tested for its effects on prostaglandin-related, laboratory models of ocular inflammation. Topically applied, BPAA inhibited arachidonic acid, but not prostaglandin E-induced increases in rabbit intraocular pressure (IOP). BPAA inhibited the IOP response to alkali burn and altered IOP changes following paracentesis. In vitro, BPAA inhibited prostaglandin production from arachidonic acid in cell-free preparations of rabbit uvea. It is suggested that BPAA may be useful for the therapy of ocular inflammatory disease.


[Chronic vegetating pyodermia in cellular immune deficiency]. The simultaneous occurrence of pyoderma gangrenosum together with other diseases, most frequently ulcerative colitis, and with disorders of the humoral immune system is known. The etiology of the individual case remains nevertheless unclear in most instances. We report on an otherwise normal patient with normal immunoglobulin levels, who showed, by use of lymphocyte differentiation and functional tests, a nearly complete lack of T-cell reactivity. A substitution with transfusions of fresh blood lead to rapid healing of the clinical picture, which so far had been resistant to long term application of antibiotics, azathioprine and topical steroids. Therefore it is recommended to look for functional defects of the T-cell system as a cause of pyoderma gangrenosum.


Behavior in the Lesch--Nyhan syndrome.

Behavior in the Lesch--Nyhan syndrome. The Lesch--Nyhan syndrome is a heritable disorder of the metabolism of uric acid in which behavioral manifestations are prominent and among the most provocative. The mutated or variant gene that determines this disorder is carried on the X chromosome. The disease is expressed exclusively in males. The molecular expression of the abnormal gene is in the completely defective activity of the enzyme hypoxanthine guanine phosphoribosyl transferase. As a result these patients overproduce uric acid and may develop early in life many of the clinical findings we associate with gout. They have in addition a variety of neurological abnormalities including mental retardation, spastic cerebral palsy, and involuntary, choreoathetoid movements. Involved patients have unusual, compulsive, aggressive behavior. Its most prominent but by no means exclusive feature is self-mutilation. The central feature in the management of this behavior is physical restraint. A number of practical procedures have been learned which facilitate the care and feeding of these patients. Promising new findings suggest that behavioral modification using extinction techniques and pharmacologic methods utilizing agents designed to increase the effective cerebral content of serotonin may each have a place in the management of behavior in this syndrome.


Anatomic variants of the cerebello-pontine cisterns and their significance in positive contrast cisternography. Far lateral extension of the cerebello-pontine cistern is a common variant which is found in greater than 50% of people. When present it may cause the internal auditory canal to be obscured during positive contrast cisternography. A technique of cisternography is described which facilitates the diagnosis of intracanalicular lesions as well as better delineating small lesions in the cerebello-pontine angle.


Immunologic considerations of large volume radiotherapy. There is clinical evidence that the immune response can increase or decrease cancer growth. 1) Cancer is increased in congenital immune deficiency states and in those given immuno-suppressants after renal transplantation; 2) there is a correlation between mononuclear cell infiltration and prognosis; 3) spontaneous regression occurs occasionally on an immunologic basis; 4) cancer specific antigens and antibodies can be demonstrated, and 5) immunotherapy can cause tumor regression. The cells that are responsible for the immune response are 1) families of thymic-derived lymphocytes, or T-cells, ("memory," "effector", "helper" or "suppressor" T-cells), and 2) bursal-derived lymphocytes, or B-cells, which on specific sensitization become either plasma cells that produce cytotoxic or blocking antibody or macrophages (A-cells) that process antigen, and may become "armed" with cytophilic antibody. The main effect of local radiotherapy is to destroy certain families of radiosensitive lymphocytes in the afferent, central and efferent components of the immune response. In Hodgkin's disease, extended field radiotherapy causes a prolonged decrease in the number of lymphocytes in the peripheral blood and a decrease in function of the cells, as determined by RNA and DNA synthesis after culture with mitogen, phytohemagglutinin. There is no correlation, however, with the values after treatment and recurrence of the disease. The effect of local radiotherapy to large volumes of tissues, on the incidence of new primary cancers, is uncertain but the majority of reports show that it is no effect.


"Cleavage" and cortical granule breakdown in Rana pipiens oocytes induced by direct microinjection of calcium. Microinjection of approximately 0.3 mug of calcium into maturing oocytes of Rana pipiens after nuclear dissolution resulted in cleavage-like constrictions, cortical granule breakdown, and formation of a structure resembling a two-cell embryo. Mg2+, Na+, or K+ did not induce any of these reactions. Larger amounts of Ca2+-induced contraction over the entire surface of oocytes or eggs, but did not induce cleavage-like constrictions; smaller amounts of Ca2+ produced either a local cortical granule reaction of the formation of one large and one small "blastomere." Furrow formation was not observed during normally induced maturation until after germinal vesicle breakdown. The location of microinjected Ca2+ determined the orientation of the resulting furrow. Ca2+-induced cortical granule breakdown occurred in full-grown nonmaturing oocytes near the site of injection. Cortical granule breakdown also occurred in maturing oocytes (after germinal vesicle breakdown but before second meiotic metaphase), but only in the blastomere containing the infected Ca2+. As expected, in mature oocytes (at second meiotic metaphase) cortical granule breakdown occurred over the entire oocyte surface, including both blastomeres. The results indicate that furrow formation and cleavage-like constrictions may be directly influenced by Ca2+, and that functional contractile elements are present near all areas of the oocyte surface. Furthermore, Ca2+ injection initiates localized cortical granule breakdown in full-grown immature and maturing oocytes.


Factors affecting pharyngeal Haemophilus influenzae type b colonization rates in children. Over 1,300 children were studied in an analysis of factors that might affect pharyngeal colonization with Haemophilus influenzae type b. Our semiquantitative methods for the culture of H. influenzae type b, consisting of inoculation of 0.001 ml of throat swab fluid on antiserum agar plates and division of the results into three grades of intensity, showed agreement as to intensity of colonization in over 80% of repeat throat cultures. Our data also suggest that throat swabs are more efficient than nasopharyngeal swabs for detecting colonization, particularly for older children. All 17 H. influenzae type b carriers found with either method were detected with throat swabs, but six had negative nasopharyngeal cultures; four of these six were lightly colonized older children. Furthermore, colony counts were apt to be higher on plates inoculated with throat swab fluids. The frequency of pharyngeal H. influenzae type b colonization in children visiting health department clinics and pediatricians' offices was low during the first 6 months of life (0.7%) but averaged 3 to 5% throughout the rest of childhood. Approximately two-thirds of the carriers were colonized at an intensity too low to be detected by standard laboratory techniques. No influence on colonization rates was found for sex, race, season, economic status, or common childhood infectious diseases such as coryza or otitis media.


Pulmonary abnormalities in intermediate alpha-1-antitrypsin deficiency. Pulmonary function studies were carried out in a group of asymptomatic nonsmoking adults with intermediate alpha-1-antitrypsin deficiency who were attending an early disease detection unit in Rochester, N. Y. All subjects were identified by specific protease inhibitor (Pi) typing. Fifteen MZ and 14 MS subjects who had never smoked cigarettes were matched by sex and age to MM controls. Spirometry, static lung volumes, and single breath-diffusing capacity were identical in all Pi type groups with no statistically significant differences noted. Maximal expiratory flow volume curves were obtained in all subjects. MZ subjects demonstrated statistically impaired maximal flow rates at 75%, 50%, and 25% of vital capacity compared to their MM controls. Total pulmonary resistance by the oscillometric method was measured at 3, 5, 7, and 9 cycle/s in the same subjects. Increased frequency dependence of resistance (defined as the difference between total pulmonary resistance at 3 cycle/s and 9 cycle/s) was observed in MZ subjects compared to MM controls. No differences were noted by this method in MS-MM pairs. The data suggest that detectable mechanical abnormalities are present in subjects with the MZ phenotype, even in the absence of established risk factors such as cigarette smoking and high air pollution.


Radioimmunoassay of the binding protein for vitamin D and its metabolites in human serum: concentrations in normal subjects and patients with disorders of mineral homeostasis. A radioimmunoassay for the binding protein for vitamin D and its metabolites (DBP) has been developed. Suitable rabbit anti-DBP antiserum was elicited after primary and one booster injection. Anti-DBP antisera, as well as antigroup-specific component antisera, produced a single, monospecific line of percipitation when reacted against purified DBP and human serum. DBP was iodinated with 125I and 125I-DBP was purified by gel filtration on Sephadex G-200. Binding of 125I-DBP by 20 nl of rabbit anti-DBP antisera was approximately 50% and was sharply competed for by 0.4-4.0 ng of DBP standard. Displacement of 125I-DBP by human serum dilutions or standard DBP gave identical curves, and only weak competition was observed with old and new world primate sera. Apo- and holo-DBP possessed indistinguishable immunoreactivity. The assay detects DBP in 1-10 nl of human serum with reasonable accuracy and with reasonable intra- and interassay precision. The mean serum concentration (+/- SEM) for a group of 40 normal adults was 525 +/- 24 mug/ml and no sex difference was observed. Higher levels were found in sera from pregnant women and women receiving oral contraceptives, and decreased concentrations were observed in premature cord and hypoproteinemic sera. No significant correlation between serum DBP levels and serum 25-hydroxycalciferol levels was found, and the DBP content of sera from vitamin D-deprived and vitamin D-treated subjects was indistinguishable from that of normal adults. DBP accounts for 6- of the alpha globulin in normal human serum. Considering the normal serum content of the parent vitamin and its metabolites to be approximately 0.1-0.2 mum, these immunoassay data confirm previous saturation analyses of human serum antiricketic sterol-binding capacity and suggest that greater than 95% of DBP circulates as the apoprotein under normal conditions.


Videometric analysis of regional left ventricular function before and after aortocoronary artery bypass surgery: correlation of peak rate of myocardial wall thickening with late postoperative graft flows. The peak rate of systolic wall thickening (pdTw/dt) in regions of the left ventricle was determined by biplane roentgen videometry in 60 patients before and a median of 14 mo after aorto-coronary bypass graft surgery. The left ventricular ejection fraction, stroke volume, and end-diastolic volume and pressure did not change significantly after surgery in the presence of patent or occluded grafts (P greater than 0.05). Statistically significant increases occurred in the peak rate of systolic wall thickening regions supplied by patent bypass grafts, and significant decreases occurred in regions with occluded grafts (P less than 0.01). Of 42 preoperatively hypokinetic regions (pdTw/dt greater than 0 less than 5.0 cm/s) supplied by a patent graft, 30 improved by an average of 2.6 cm/s after operation; 18 returned to normal. Failure of 24 hypokinetic regions to improve to normal was associated with myocardial infarction in 11 or with late postoperative graft blood flows of less than 60 ml/min measured by videodensitometry, in 10. All seven preoperatively akinetic (pdTw/dt=0) or dyskinetic (pdTw/dt less than 0) regions did not improve after the operation despite the fact that, in five of the seven, coronary bypass flows were over 60 ml/min. All eight preoperatively hypokinetic regions supplied by coronary artery graft flows of less than or equal 40 ml/min failed to improve to normal after operation. All nine preoperatively hypokinetic regions supplied by coronary artery graft flows of over 60 ml/min improved to normal after surgery. Late postoperative coronary artery bypass graft flows, the functional status of the myocardium, the status and distribution of the native coronary circulation, and decreased regional function elsewhere in the ventricle must all be considered when regional left ventricular function is interpreted.


Milk fever in dairy cows. VII. Effect of continuous vitamin D feeding on incidence of milk fever. Feeding of 20 to 30 million units of vitamin D for 3 to 8 days prepartum previously prevented about 80% of expected milk fever cases in mature Jersey cows with histories of milk fever. In this experiment vitamin D was fed continuously through the year via 32,000 units of vitamin D added to each .455 kg of concentrate fed (approximately 100,000 to 580,000 units/day). Milk fever incidence was measured after 5 yr of age (third calf) at 43 parturitions in mature Jersey cows with histories of milk fever and at 139 parturitions in mature Jersey cows with no previous milk fever. Continuous feeding of vitamin D reduced incidence of milk fever in cows with previous milk fever from 60% in the controls to 26.1% in the group fed vitamin D. In the cows with no milk fever previously, feeding vitamin D did not reduce incidence of milk fever (controls 23.7%, vitamin-D-fed 28.3%).


Massive postoperative hemorrhage from hepatic artery erosion. A 66-year-old male patient who had undergone repeated operations for peptic ulcer disease involving the right upper abdominal quadrant, developed cholecystitis with calculous obstruction of the common bile duct. The gallbladder was removed. Later, an operation was performed for removal of a residual stone from the common duct. At this time an anomalous arterial structure was noted about the duct. Hemorrhage occurred ten days postoperatively, and the anomalous hepatic artery was found to be eroded. The bleeding was controlled. During the succeeding two weeks there were four episodes of bleeding (involving erosion of the hepatic artery and adjacent tissues), three of which were controlled. The fourth episode ended in the death of the patient from exsanguination secondary to bleeding from stress ulcers in the gastric remnant. At no time did the laboratory data unequivocally indicate an abnormality of blood coagulation. Erosion of the anomalous cystic artery apparently precipitated the fatal chain of events.


Evidence for separate subpopulations of B cells responding to T-independent and T-dependent immunogens. BALB/c spleen cell cultures were double-challenged with optimal doses of TNP-T independent and TNP-T-dependent immunogens. The anti-hapten response was additive or synergistic in double challenged cultures compared to the response in cultures challenged with either immunogen alone. A high, suppressive dose of either immunogen did not prevent the response to an optimal dose of the second immunogen in the same culture. Spleen cell cultures were also double challenged with combinations of two T-dependent or two T-independent immunogens. Addition or synergy in the anti-hapten response only occurred when the double challenge consisted of a mixutre of T-dependent and T-independent immunogens. These results are consistent with the proposed B1B2 classification of B cell subpopulations.


Membrane expression of Thy-1,2 and GM1 ganglioside on differentiating T lymphocytes. C3H mouse bone marrow cells were separated by discontinuous BSA gradient centrifugation. Marrow cells from the 17 to 19%, 19 to 21% to 23%, 23 to 25%, 25 to 27% interfaces and the cell pellet were treated with thymic factor (TF) or with Vibrio cholerae neuraminidase (VCN), followed by anti-Thy-1,2 and anti-GM1 ganglioside antisera. Antigens determined by anti-Thy-1.2 or anti-GM1 were expressed either with TF or VCN within a 30-min incubation. Cells expressing these antigens after VCN or TF treatment were concentrated in the 19 to 21% layer and the 21 to 23% layer whereas there was little or no antiserum cytotoxicity in the other layers. A small amount of Thy-1.2 of GM1 could be detected by cytotoxicity on 19 to 21% layer marrow cells within 15 min of either TF or VCN treatment. Treatment of 19 to 21% layer cells with TF or VCN had no effect on anti-H-2 cytotoxicity. Pretreatment of TF-treated marrow cells with cholera toxin or choleragenoid (which bind cell surface GM1) abrogated the cytotoxicity of anti-Thy-1.2 or anti-GM1 without affecting anti-H-2 cytotoxicity. Pretreatment of Thy-1.2 positive C3H thymocytes with cholera toxin or choleragenoid greatly reduced the cytotoxicity of anti-Thy-1.2 antisera without affecting the cytotoxicity of anti-H-2 antisera. Nude mouse splenocytes, after treatment with VCN or TF, were susceptible to the lytic action of anti-Thy-1.2 and anti-GM1. The possibility that nonspecific autologous antibodies were responsible for anti-Thy-1.2 cytotoxicity toward VCN-treated marrow cells was eliminated because anti-Thy-1.2 was not cytotoxic for VCN treated AKR (Thy-1.1) marrow cells.


Studies of surface immunoglobulins on human B lymphocytes. II. Characterization of a population of lymphocytes lacking surface immunoglobulins but carrying Fc receptor (SIg-Fc+ cell). Surface markers and organ distributions of B lymphocytes bearing surface immunoglobulins (SIg B) and lymphocytes lacking SIg but carrying Fc receptors (SIg-Fc+), and the variations of both the cell populations in the autoimmune diseases were studied. Peripheral blood lymphocytes contained approximately 9% SIg B and 22% SIg-Fc+ cells. Both the populations of SIg B and SIg-Fc+ cells were included in the cells that formed no E-rosettes. When previously treated with neuraminidase, however, the SIg-Fc+ cells formed E-rosettes, whereas SIg B showed no ability to form rosettes. C receptors as detected by human complement were found in the large portions of SIg B but in a minority of the SIg-Fc+ cell population. The tissues that contained the highest concentrations of SIg B and SIg-Fc+ cells were the spleen and peripheral blood, respectively. In the patients with an autoimmune disease in active stages SIg B cells in the peripheral blood markedly increased and, reversed, SIg-Fc+ cells decreased; in contrast, in remission of the diseases SIg B cells dropped down and SIg-Fc+ increased up to their normal levels. A possible relationship between the SIg B and SIg-Fc+ cells in regard to the differentiation of B lymphocytes was presemed.


Complement receptor lymphocytes in the rabbit. II. The effect of immunization on the complement receptor lymphocyte population in the craining lymph node. In an attempt to determine the effect of immunization on the size and nature of the complement receptor lymphocyte (CRL) and surface immunoglobulin (SIg)-bearing lymphocyte (B cell) populations, rabbits were injected with 1 mg alum-precipitated keyhole limpet hemocyanin and the cells of the draining popliteal lymph nodes were examined at various intervals for these populations. Comparison to unimmunized control lymph nodes revealed that immunization resulted in an approximate doubling of the percentages of both CRL and B cell populations by the peak of the response, while the cell number increased about 7-to 8-fold. Kinetic analysis of the response indicated that the percentages of CRL and B cells increased essentially in parallel beginning within 40 h after immunization. Simultaneous assay of complement receptor (CR) and SIg disclosed that CRL remained a B cell subpopulation throughout the response. To determine whether cell division could account for the increases in total cell number and per cent CRL and B cells, immunized rabbits were given 3H-thymidine in vivo, and the fraction of labeled (dividing) cells was determined by autoradiography. Results of such experiments were consistent with the possibility that the early increase in cell number and the increase in CRL and B cell percentages were a result of trapping and/or homing of lymphocytes migrating from the peripheral blood. A significant amount of lymphocyte division occurred during later stages of the response.


Induction or suppression of B cell proliferation and differentiation by phytohemagglutinin or concanavalin A in mouse spleen cell cultures. Cultures of mouse spleen cells or various mixtures of mouse T and B cells were stimulated with PHA or Con A, and the T, B, or plasmablast nature of the transformed cells, was determined by immunofluorescence 2 to 4 days later. The lectins enhanced B cell proliferation and plasmablast differentiation ("helper" effect) provided one of the following conditions was fultilled: a) suboptimal doses of lectin were used, b) cultures were performed at low cell concentration, c) cultures were made of spleen cells containing a small percentage of T cells, d) the cultures contained a mixture of T-depleted spleen cells and T cells rendered unable to proliferate by irradiation. In contrast, cultures performed with 1.5 10(6) or more spleen cells/ml and optimal doses of lectin contained almost exclusively T blasts, as did cultures stimulated in the same conditions with both PHA and LPS. This last observation idicates the existence of a lectin-induced "suppressor" effect, since LPS, a B cell mitogen, induces, in the absence of PHA, a marked B cell proliferation and differentiation into plasmablasts. These helper and suppressor effects were entirely mediated by T cells, since they were not observed in spleen cell cultures depleted in T cells by anti-thets + C. Analysis of the cultures by immunofluorescence and radioautography after pulses of 3H-thymidine showed that these antagonistic effects could be related to the number of T blasts present in the culture and to their proliferative behavior. Heoper effect is observed in cultures containing a relatively low number of T blasts (or none in cultures made with irradiated T cells), whereas suppressive effect is observed in cultures containing a high number of T blasts, a large proportion of them having left the proliferative cell cycle. It is proposed that when a critical concentration of T blasts is reached ("saturation density"), further proliferation and differentiation is prevented, resulting in a suppressive effect on the generation of plasmablasts. The helper effect of lectin-activated T cells seems to be exerted on a subpopulation of B cells which was, at least in part, already proliferating in vivo, and to result in a polyclonal IgM plasmablast differentiation.


Role of T lymphocytes in adjuvant arthritis. I. Evidence for the regulatory function of thymus-derived cells in the induction of the disease. Rats of Wistar King Aptekman (WKA) were thymectomized at 4 weeks of age and injected with Mycobacterium tuberculosis wax D 6 weeks after the operation to induce adjuvant arthritis. The development of this disease was strikingly enhanced by this treatment. Further experiments showed that a 4- to 6-week interval between thymectomy and wax D injection was necessary to show the enhancing effect. Such enhancement by thymectomy was also shown in rats of another strain, Sprague-Dawley (SD). The enhancing effect of thymectomy was abolished when thymocytes of normal syngeneic rats were transferred to thymectomized rats 7 days before the wax D inoculation. Furthermore, severe arthritis was also produced in WKA rats that were pretreated with low dose (200 R)  whole body irradiation, but not in those treated with higher doses (400 to 700 R). These results seem to indicate that the enhancing effect is brought about by selective depletion of a certain population of T lymphocytes. The population depleted may be thymus dependent, short-lived and radiosensitive, the properties of which agree whith those known for suppressor T lymphocytes. Thus, it appears that thymus-derived cells could normally exert a regulatory effect on the development of adjuvant arthritis which might render these rat strains relatively less susceptible to this disease.


Electrophoretic separation of radioisotopically labeled mouse lymphocytes. A new preparative method is described for the separation and direct electrophoretic comparison of the distribution profile of radioisotopically labeled cells. The cell populations to be compared are labeled separately with different radioisotopes (e.g., 51Cr and 99mTc) and mixed. The cell mixture is subjected to density gradient electrophoresis and the distribution of each radioisotope in the collected fractions is ascertained by differential gamma counting. In the case of uniform label uptake by the cells in each distribution, the radioactivity counts represent cell frequency. Nonuniform labeling allows useful comparisons only in terms of relative mobility distribution. Model experiments performed with 51Cr- and 99mTc-labeled mouse thymocytes and spleen cells are in general agreement with mobility distribution results obtained previously by free flow electrophoresis.


Genetic control of immune responsiveness to poly (LPro)--poly (LLys)-derived polypeptides by histocompatibility-linked immune response genes in the rat. In rats responsiveness to branched synthetic polypeptides carrying a Pro--L backbone, such as (T,G)-Pro--L or (Phe,G)-Pro-L and to Pro--L itself is controlled by Ir genes which are linked to the major histocompatibility genes. The level of antibody production to these polypeptides does not fall into strict high or low responder categories but covers the range in between. (T,G)-pro--L and Pro--L elicit a very similar response pattern which, however, differs from that obtained with (Phe,G)-Pro--L. Anti-(T,G),Pro--L antibodies do not cross-react with (T,G)-A--L, but do so extensively with Pro--L. Anti-(Phe,G)-Pro--L antibodies show cross-reactivity to (Phe,G)-A--L only when the antibody-producing strain is a high responder to (Phe,G)-A--L. These results when considered in view of data obtained in mice on genetic control of the immune response to (T,G)-Pro--L suggest that at least two unlinked Ir genes are involved in controlling anti-Pro--L responsiveness.


Cytotoxic macrophages: a rapid nonspecific response to viral infection. In vitro and in vivo assays have been developed to study the relative contributions of various types of immune cytolysis in the destruction of infected cells after Semliki Forest virus infection of BALB/c mice. Highly cytotoxic activated macrophages, not specific for the infecting virus, appear on day 1, peak on day 2 to 3, and disappear within a week. Specifically sensitized T cells appear around day 3, peak on day 6, and disappear within a month. Cytotoxic antibody appears on day 4 and reaches high titers by day 8. Immune spleen cells greatly reduce the yield of virus from cultured cells. Infected cells rapidly disappear after transfer to infected animals.


Functional heterongeneity among the T-derived lymphocytes of the mouse. VI. Memory T cells stored in the T2 subpopulation. Memory T cells were demonstrated in mice which were the precursors for cells causing sheep red blood cell-(SRBC) specific helper activity in the in vitro response of spleen cells to trinitrophenyl (TNP)-SRBC. These memory cells were distinguished from the effectors of helper activity generated during the primary response and were shown to belong to the T2 subpopulation of peripheral T cells on the basis of 1) their rapid response to a challenge with SRBC, 2) their long lifetime, and 3) their sensitivity to small in vivo doses of anti-thymocyte serum.


Mouse B and T lymphocyte responses to purified timothy pollen antigens in vitro. Purified populations of splenic B and T lymphocytes from LAF mice immunized with a crude extract of timothy pollen (WST) responded specifically to pollen antigens in an in vitro lymphocyte transformation system. The peak lymphocyte transformation response occurred 5 days after a secondary immunization and was the result of T-B cell cooperation in vitro. Wtih two purified pollen antigens as in vitro stimulants we were able to define at least two antigen-specific population of B cells and one population of T cells. These results were confirmed by inhibition studies with a monovalent hapten from WST, Antigen D.


Subpopulations of chicken B lymphocytes. Immunoglobulin-synthesizing cells from the spleen and bursa were fractionated by the 1 X G sedimentation velocity technique and characterized by their ability to synthesize immunoglobulin and by staining with fluorescent anti-light chain chain. Four subpopulations of immunoglobulin-synthesizing cells were identified. In the bursa, slowly sedimenting (S 2.3 mm/hr) and rapidly sedimenting (S greater than 3.5 mm/hr) subpopulations with surface immunoglobulin were present; in the spleen, a slowly sedimenting (S 2.3 mm/hr) subpopulation with surface immunoglobulin and plasma cells (S greater than 3.5 mm/hr) with large concentrations of intracellular immunoglobulin existed. The subpopulations differed most markedly in their ability to synthesize immunoglobulin (cpm Ig synthesized/10(6) Ig-positive cells); the rates of immunoglobulin synthesis were in the ratio 1:2:1:900. The slowly sedimenting B cells from the spleen and both subpopulations of B cells from the bursa released small amounts of immunoglobulin into the culture media, whereas, the plasma cells released immunoglobulin at a rate as much as 3700 times greater. Bursal B cells could be further distinguished from splenic B cells by a greater amount of DNA synthesis.


Fractionation of cytotoxic cells from tumour-immune rats on derivatized collagen gels. Incubation at 37 degrees C of spleen cells from rats, immunized with Gross virus-induced lymphoma, on collagen gels coated with membrane extracts of lymphoma cells yielded two different cell populations. The non-adherent cells showed greatly diminished cytotocicity for lymphoma cells compared with unfractionated spleen cell suspensions, whilst the specifically adherent cells which were recovered after collagenase digestion of the gels, showed enrichment of cytotoxic activity.


Studies of mouse-brain-associated T-lymphocyte antigens. Xeno- and alloantigens shared by mouse brain, thymocytes and thymus-derived lymphocytes were isolated from brain by butanol extraction, followed by Sephadex G-200 column chromatography and preparative polyacrylamide gel electrophoresis, both in the presence of SDS. The antigenic activity, as tested by inhibition of complement-mediated cytotoxicity using a xenogeneic antiserum directed against T cell-specific antigens (anti-MTLA), was enriched 50-fold during the purification procedure. Xenogeneic and allogeneic antisera which were specific for mouse T-lymphocytes, could be obtained by immunization of rabbits and mice with the partially purified antigens.


Serologic responses of children with meningitis due to Haemophilus influenzae type b. The immunologic responses of 100 children hospitalized with meningitis due to Haemophilus influenzae type b were measured by the bactericidal antibody assay (BAA) and radioimmunoassay (RIA) for detection of antibody. Short-term (14-17 days after onset of illness) responses were detected by RIA alone in 20 children, by BAA alone in six, and by both tests in 23. The more sensitive RIA detected 20 children who would have been labeled "immunologically unresponsive" had only BAA been used. The magnitude of the antibody response was clearly related to age. Of 26 children with no immediate antibody response, 11 still had no rise in titer of antibody when restudied two to 20 months later; the remaining 15 had subsequent increases in titer. Nine of 10 children who showed an immediate antibody response remained positive when additional blood smaples were taken two to 18 months later. Over half of the children initially unresponsive to H. influenzae type b meningitis subsequently developed specific antibodies. The remainder, who failed to acquire detectable antibodies at either the acute stage of illness or late in convalescence, deserve further investigation as to the nature of their immunologic hyporesponsiveness to H. influenzae type b meningitis.


Amikacin in the treatment of gram-negative bronchopulmonary infections. Six patients with acute gram-negative bronchopulmonary infection were treated with amikacin (15 mg/kg per day) administered intramuscularly in two equal doses at 12-hr intervals for 10-13 days. Two patients had underlying nonspecific pulmonary disease, two had advanced bronchocarcinoma, and two had extensive bronchiectasis (due to chronic aspergillosis in one patient). The pathogens were Pseudomonas aeruginosa in three patients, and Haemophilus influenzae, Klebsiella ozaenae, and Enterobacter cloacae each in one patient. Five patients recovered completely, with resolution of fever and other acute symptoms and elimination of the causative organism from sputum cultures. A moribund patient with advanced metastatic bronchocarcinoma died two days after the treatment with amikacin had been completed; the last specimen of sputum was still positive for P. aeruginosa. Tests of liver and renal function and blood counts revealed no abnormaltities. Complete audiometric survey showed no hearing loss. Nystagmography revealed reversible, lessened caloric response in some patients. Amikacin was well absorbed from the site of intramuscular injection. Levels of amikacin in serum varied among the subjects and, in some cases, for individual patients on different days.


A reappraisal of cellular labeling with radiochromium salts. The present studies demonstrate that, while erythrocytes preferentially bind hexavalent 51Cr, leukocytes bind the trivalent form with equal avidity. Red cells may be saturated with label in contrast to the binding of 51Cr to leukocytes, which may be described by first-order kinetics. Hexavalent label is rapidly reduced in leukocytes in contradistinction to the finding in red cells. The nonreutilization of label by erythrocytes and leukocytes is not due to inappropriate valency but to protein binding of the label. Unlike nonleukemic cells, leukemic leukocytes elute a large fraction of their label. These factors have been largely overlooked in many previous clinical studies using labeled cells, and they are of sufficient quantitative importance to compromise seriously the validity of numerous earlier reports.


Role of B lymphocytes in cell-mediated immunity. I. Requirement for T cells or T-cell products for antigen-induced B-cell activation. Although B lymphocytes can be triggered by B-cell mitogens and by certain other molecules to produce lymphokines, they do not produce lymphokines when stimulated with specific soluble protein antigens. We have investigated whether T-cell help would enable B cells to produce lymphokines when activated by antigens. Addition of small numbers of T cells to B-cell cultures resulted in significant production of a monocyte chemotactic factor. T cells could be replaced by supernates of antigen-stimulated T cells, demonstrating both that the chemotactic factor was B-cell-dervied and that T-cell help was mediated by a soluble factor. Although the T-cell factor was nonantigen specific, B-cell activation required the presence of both antigen and T-cell factor. Thus, it appears that although dependent upon T cells, B lymphocytes may play an important role in amplification of cell-mediated immune responses.


Regulatory mechanisms in cell-mediated immune responses. IV. Expression of a receptor for mixed lymphocyte reaction suppressor factor on activated T lymphocytes. Suppression of the mixed lymphocyte reaction (MLR) by a soluble factor produced by alloantigen-activated spleen cells requires genetic homology between the factor-producing cells and responder cells in MLR. The ability of lymphocytes used as MLR responder cells to adsorb MLR suppressor factor was tested to investigate the expression of a receptor structure for suppressor molecules. Normal spleen or thymus cells had no effect on suppressor activity. Concanavalin A (Con A)-activated thymocytes, however, effectively removed suppressor activity, suggesting that the receptor is expressed only after activation and is not present or not functional on resting cells. Significantly neither phytohemagglutinin- nor lipopolysaccharide-activated lymphoid cells absorbed the factor. Furthermore, only Con A-activated thymocytes demonstrating genetic homology with the cell producing suppressor factor for H-2 regions to the right of I-E were effective absorbants. Alloantigen-stimulated spleen cells syngeneic to the suppressor cell also removed suppressor activity. These data support an hypothesis that subsequent to stimulation in MLR, T lymphocytes express a receptor, either through synthesis or alteration of an existing molecular structure, which then provides the appropriate site for interaction with suppressor molecules.


Allogeneic carrier-specific enhancement of hapten-specific secondary B-cell responses. We have analyzed the capacity of carrier-specific T cells to enhance the immune response of hapten-specific secondary B cells which do not share genes in the H-2 complex with the T cells. For this analysis we have used the in vitro splenic focus technique which allows assessment of monoclonal responses of B cells isolated in splenic fragment cultures of irradiated reconstituted carrier primed mice. A previous report from this laboratory demonstrated that syngeny in the I region of the H-2 complex was necessary between collaborating hapten-specific primary (nonimmune) B cells and carrier-specific T cells for responses yielding IgG1 but not IgM antibody. These findings lead up to postulate that the expression of I-region gene products on the surface of primary B cells and I-region syngeny with collaborating carrier-specific T cells were essential elements in the triggering events leading to IgG1 synthesis by primary B cells. The results presented in the present report indicate that, unlike primary B cells, the majority of secondary B cells can be stimulated to produce IgG1 antibody in carrier-primed allogeneic recipients. Although the enhancement of secondary IgG1 responses is slightly greater with syngeneic T cells, the allogeneic collaborative interaction requires both carrier priming of recipient mice and stimulation with the homologous hapten-carrier complex and thus appears to be specific. These findings clearly discriminate secondary from primary B cells and indicate that the mechanism of stimulation of secondary B cells to yield IgG1-producing clones differs fundamentally from the stimulation of primary B cells in that the requisite for I-region syngeny is obviated.


Isoantiserum-augmented development of lymphocyte-mediated cytotoxicity. Passive administration of anti-P-814 isoantiserum (IS) with P-815 mastocytoma was shown to augment the development of T-lymphocyte-mediated cytotoxicity (LMC). The LMC activity augmented by IS was specific to immunizing tumor cells, but required the simultaneous administration of P-815 tumor cells and anti-P-815 serum, suggesting that the antigen-antibody complex is involved in the development of specific cytotoxic T cells. The serum component responsible for augmented development of LMC activity appeared to be IgM in that the augmenting activity fractionated in the void volume of a G-200 Sephadex column and appears very early after immunization. Our experimental results suggest the development of specific T-cell cytotoxicity can be directly regulated by specific IgM antibodies.


Role of self-carriers in the immune response and tolerance. I. B-cell unresponsiveness and cytotoxic T-cell immunity induced by haptenated syngeneic lymphoid cells. Normal spleen cells cultured with TNP-modified syngeneic spleen cells fail to mount an anti-TNP PFC response to TNP-ficoll or TNP-red blood cells,but go on to generate cytotoxic T cells directed at hapten-modified H-2.These results suggest that hapten-modifeid spleen cells may differentially induce B-cell tolerance and T- (Ly 2,3) cell immunity. The differential response to modified self by lymphocyte subpopulations is discussed.


Fractionation of human T lymphocytes on wheat germ agglutinin-sepharose. T cells from human peripheral blood was purified by fractionation on columns charged with human immunoglobulin and rabbit anti-human immuno-globulin. When assayed with 125I- or fluorescein isothiocyanate-labeled wheat-germ agglutinin (WGA), a weakly binding and a strongly binding subpopulation could be distinguished. These T-cell subpopulations were fractionated on columns charged with WGA, convalently bound to Sepharose 6MB. The cells responding to the mitogens leukoagglutinin from Phaseolus vulgaris and concanavalin A were enriched in the strongly binding subpopulation (approximately 20% of the T cells) while they were depleted from the weakly binding subpopulation.


An oral test of pancreatic chymotrypsin activity using N-acetyl-L-tyrosyl-p-amino-benzoic acid in rats. After a single dose of up to 50 mg/kg body weight (optimal amount for the test) of p-aminobenzoic acid or the corresponding amount of synthetic chymotrypsin substrate N-acetyl-L-tyrosyl-p-aminobenzoic acid, the urinary excretion of p-aminobenzoic acid in rats increases approximately linearly during 24 h. Higher doses do not cause any further significant increase in the amount of urine excreted p-aminobenzoic acid. Compared with a 24 h collecting period, 82% of p-aminobenzoic acid and 77% of p-aminobenzoic acid from N-acetyl-L-tyrosyl-p-aminobenzoic acid were excreted in the urine during the first 6 h after oral application of these substances. Therefore it is sufficient for practical purposes to determine p-aminobenzoic acid in 6 h urine samples. Stimulation of pancreatic secretion by application of 2.5 U of pancreozymin and 2.5 U of secretin failed to bring about significant increase in the cleaved p-aminobenzoic acid in the urine.


Strophanthidin-sensitive sodium fluxes in metabolically poisoned frog skeletal muscle. Strophanthidin-sensitive and insensitive unidirectional fluxes of Na were measured in fog sartorius muscles whose internal Na levels were elevated by overnight storage in the cold. ATP levels were lowered, and ADP levels raised, by metabolic poisoning with either 2,4-dinitrofluorobenzene or iodoacetamide. Strophanthidin-sensitive Na efflux and influx both increased after poisoning, while strophanthidin-insensitives fluxes did not. The increase in efflux did not require the presence of external K but was greatly attenuated when Li replaced Na as the major external cation. Membrane potential was not markedly altered by 2,4-dinitrofluorobenzene. These observations indicate that the sodium pump of frog skeletal muscle resembles that of squid giant axon and human erythrocyte in its ability to catalyze Na-Na exchange to an extent determined by intracellular ATP/ADP levels.


Neuroelectric blocking factors in human and animal sera evaluated using the isolated frog spinal cord. The effects of sera from guinea-pigs with experimental allergic encephalomyelitis (EAE) and multiple sclerosis (MS) patients were evaluated and compared with the activity of control sera using the isolated frog spinal cord. Ventral root responses (VRR) were recorded during supramaximal ipsilateral dorsal root stimulation in the presence and absence of 25% serum. In control experiments with normal human and guinea-pig sera we observed a consistent, reversible increase in VRR averaging 20% and 17% respectively, and in no case was any significant decrease produced. In contrast, sera from EAE guinea-pigs 12 to 19 days after injection produced an equally rapid, reversible decrease in VRR. The decrease averaged 36% and was highly significant (P less than 0.0001) relative to controls. Similarly, sera from MS patients on the average decreased the VRR by 26%, and this again was significant compared with controls (P less than 10(-6).


Mechanism of the inverted supinator reflex. A clinical and neurophysiological study. The inverted supinator reflex sign was analysed in two patients by recording the response of the finger flexor muscles of the forearm to tapping of the styloid process of the radius before and after procain infiltration of these muscles. It was found that the response persisted despite the block. The findings support the concept that the underlying mechanism is increased alpha motoneurone excitability below the level of the lesion; however, a possible contribution of the dynamic muscle spindles could not be excluded.


A potential error in the quantitation of fecal blood loss: concise communication. Chromium-51-labeled red cells were used to quantitate fecal blood loss in a patient with chronic upper gastrointestinal hemorrhage. On Day 1, the stool guaiac was positive but the blood loss indicated by 51Cr was less than 1 cm3. Blood loss in the stool by 51Cr did not become significant until Day 3, when it measured 23 cm3. The failure to detect abnormal blood loss on Day 1, and probably on Day 2, appears to be due to a long intestinal transit time from a proximal bleeding site. The problem of slow intestinal transit is not uncommon and could lead to a false-negative study or falsely low estimates of fecal blood loss. This problem could be minimized by beginning stool collection on Day 3 or by delaying stool collection until the appearance in the stool of an oral nonabsorbable marker swallowed when the 51Cr-tagged red cells are injected.


Proctosigmoidoscopy and rectal biopsy in infants and children. The medical records of all patients ages 0 to 21 years who underwent proctosigmoidoscopy and/or rectal biopsy over a 27 month period of time were reviewed to determine the efficacy and safety of these procedures in pediatric patients. One hundred twenty-one patients underwent proctosigmoidoscopy; 91 of these also had rectal biopsies. Median age was two years; 21% were less than six months and 8% less than one month of age. Depth of examination was 10 to 15 cm in most patients greater than 10 years of age. Induced friability was the most frequently observed mucosal abnormality. Abnormal findings were almost always present in patients with bloody diarrhea and were quite common in those with rectal bleeding, but less common in those with chronic diarrhea and abdominal pain. Colitis of various causes was the most common cause of blood in the stool; anal fissures were found in only four of 23 patients with rectal bleeding. Both proctosigmoidoscopy and rectal biopsy were needed to exclude the presence of colitis. Mobidity was 0% with proctosigmoidoscopy and 0.34% with rectal suction biopsy.


Changes in membrane currents in bullfrog atrium produced by acetylcholine. 1. A double sucrose-gap voltage-clamp technique has been used to study the effects of acetylcholine on the membrane currents in atrial trabeculae of the bullfrog, Rana catesbeiana. 2. The second, or slow inward (Ca2+/Na+) current, was found to be markedly reduced by concentrations of acetylcholine greater than approximately 2-0 X 10(-8)M. The resulting decrease in net calcium entry provides a straightforward explanation for the negative inotropic action of acetylcholine in atrial muscle. 3. Measurements of membrane resistance near the resting potential showed that relatively high doses of acetylcholine (approximately 10(-7) M) decrease membrane resistance by about twofold. This effect is shown to be the result of an increase in a time-independent background current which appears to be carried mainly by potassium ions. 4. Using appropriate pharmacological techniques, it has been possible to demonstrate: (i) that the peak slow inward current is reduced to about half its initial value before any significant increase in background current occurs; (ii) that even when a sufficient dose of acetylcholine to produce an increase in background current is used, the background current shows inward-going rectification and cannot account for the observed reduction in the slow inward current. 5. No consistent change was observed in the degree of activation of the time-dependent outward membrane currents after application of concentrations of acetylcholine which produced large decreases in the peak slow inward current. 6. These results are discussed in relation to previous electro-physiological and radioisotope studies of the mechanism of the negative inotropic effect of acetylcholine in cardiac muscle.


Action of vinblastine on the spontaneous release of acetylcholine at the frog neuromuscular junction. 1. Vinblastine induces reversible changes of the spontaneous release of acetylcholine (ACh) at the frog neuromuscular junction as characterized by the appearance of giant potentials. These large potentials occur soon after soaking the muscle in vinblastine and are not consequent to a large increase in the frequency of spontaneous release. Their number, relative to the total number of spontaneous potentials, increases with the duration of soaking. 2. Large potentials appear even in the presence of tetrodotoxin or in low Ca2+-high Mg2+ Ringer solution. 3. Following vinblastine treatment,the amplitude histogram of spontaneous potentials recorded from a number of fibres display an evident periodicity with peaks occurring regularly at simple multiples of the model amplitude of the unitary potentials. It is suggested that giant potentials are produced by the release of preformed pluriquantal packets of ACh. 4. Comparison of the amplitude distribution of spontaneous potentials and end-plates potentials show that only an insignificant number of large quanta are released by nerve stimuli. The absolute frequency of giant potentials does not markedly change when spontaneous discharge is accelerated by hypertonic solution. 5. The mechanism by which vinblastine induces the appearance of giant potentials is discussed.


Current-voltage relationships in the crystalline lens. 1. Electrical coupling between the cells of the crystalline lens of the frog eye was studied using two intralenticular micro-electrodes, one to pass current and one to record potential. In most experiments, both electrodes were placed just inside the posterior surface of the lens at a depth of approximately 200 mum from the surface. Step functions of current were applied and the time course of the resulting change in voltage was measured at many different electrode separations. 2. The voltage change has both a fast component, which occurs only locally in the region close to the current passing micro-electrode, and a slow component, which is spatially uniform, independent of distance from the current micro-electrode. 3. This behaviour is predicted by an electrical model of a single large spherical cell, and so that model can be used to analyse our data. 4. The resistivity of the lens 'interior' (both cytoplasm and coupling resistivity) is 625 omega cm; the resistance of the lens 'membrane' is 2751 omega cm2. 5. The data and analysis help to reconcile discrepancies between previous measurements of the electrical properties of the lens and show clearly that there is substantial electrical coupling from cell to cell. The method should allow investigation of the role of electrical coupling in cataract formation in the crystalline lens.


The role of some small peptides in the transfer of amino nitrogen across the wall of vascularly perfused intestine. The characteristics have been investigated of the transfer into the vascular bed of L-leucine and glycine from free amino acids or peptides in the intestinal lumen of Rana pipiens. Over the concentration range 0-5-10 mM the transfer of L-leucine is but little affected by the presence of equimolar concentrations of glycine but the transfer of glycine, in contrast, is greatly inhibited by the presence of L-leucine. 2. With glycyl-L-leucine in the intestinal lumen, the rate of transfer of glycine into the vascular bed is much greater than from the mixture of the two amino acids and is equal to that of the L-leucine. From L-leucyl-glycine the rates of transfer of leucine and of glycine are also higher than from the mixture of the two amino acids but the rate of transfer of glycine is somewhat lower than that of leucine. There is no evidence of the presence of the dipeptides in the effluent from the portal vein. 3. When the peptide glycyl-L-leucine is added to the lumen in the presence of 10 mM concentrations of the free amino acids, additional amounts of L-leucine and of glycine are transferred in approximately equimolar quantities into the vascular bed. This additional transfer exhibits saturation with respect to concentration of peptide in the intestinal lumen. An additional transfer of amino acids was also found when L-leucyl-glycine was added to the lumen in the presence of saturating concentrations of the two amino acids. 4. Evidence is presented that the presence of the dipeptides in the intestinal lumen had little effect on the transfer of free amino acids from the lumen into the vascular bed. Although the transfer of free amino acids from the lumen into the vascular bed is significantly, but not completely, abolished when the Na in the intestinal lumen is replaced by K, the transfer of the amino acids from the dipeptides is but little affected. 5. The findings are discussed in relation to the view that the dipeptides are transported into the mucosal epithelium by a process that is distinct from those promoting uptake of the individual amino acids. Being completely hydrolysed, there is no evidence for an accumulative uptake of the peptides; it is suggested that this may be related to the fact that the peptide uptake occurs in the absence of Na in the intestinal lumen.


Studies in the neurophysiology of learning: X. Phase coherence of brain potentials as index of alertness and orienting response in the rat. With use of electrical potentials derived from chronically implanted electrodes in the rat's forebrain to trigger a computer of average transients (CAT), averaged traces were obtained which gave characteristic undulations under conditions of arousal and alertness (N = seven albino rats--six male and one female). Under conditions of extreme adaptation to the environmental condition, the averaged undulations disappeared. The undulations were characterized by a period of 125-150 milliseconds and occurred in trains of .3-.5 second. The parameters of the waves were constant for a variety of novel environmental conditions, but the waves vanished in those conditions that appeared to greatly stress or terrify the rat. The undulations so obtained were conditionable to initially neutral stimuli in both the forward and backward directions.


Lymphocytes forming rosettes with sheep erythrocytes in metastatic pleural effusions. Percentages of lymphocytes forming rosettes with sheep erythrocytes at 29 degrees C were determined in 10 cancer patients with metastases to the pleural cavity. Compared with normal controls, the patients showed a decreased proportion of rosette-forming cells (RFC) in the peripheral blood. The same patients had elevated levels of RFC in their metastatic pleural effusions. However, 2 patients with benign diseases had normal levels of RFC in their peripheral blood and pleural transudates. These observations suggested that in cancer patients some T-cells might migrate from the peripheral blood and accumulate in sites of tumor infiltration such as the pleural cavity.


Regulatory variants for the expression of H-2 antigens. II. Level of discrimination of cell-mediated cytolysis. We used alloantigenic variants to analyze whether killer T-cells and antibody in the presence of complement (two effector components of the immune system) recognized the same determinants on target tissues. The variants were selected from mouse myeloma cells by immune selection and differed from the parent cell in that they did not express H-2 antigens. The results suggested that both killer T-cells and humoral effectors recognized the same determinants, though the killer T-cells may have been more sensitive either in recognizing these determinants or mediating lysis.


The probability of closure in aortocoronary vein bypass grafts. A study of 540 patients who had the aortocoronary vein bypass operation (from several transplant teams) showed that grafts within a patient closed independently of one another during the first-two year postoperative period. For patients with a single vein graft, there was a 23% chance of closure during the first two year postoperative period. The chance that all grafts would close during this period for patients with two grafts was 5%, and it was 1% for patients with three grafts. The knowledge of these closure probabilities may be used by the surgeon when planning the optimal surgical maneuver.


Antihemophilic factor concentrate therapy in von Willebrand disease. Dissociation of bleeding-time factor and ristocetin-cofactor activities. Treatment of von Willebrand disease with two plasma antihemophilic factor (AHF) concentrates, cryoprecipitate and glycine-precipitated AHF, was compared. Both concentrates were equally effective in immediately raising the plasma levels of factor VIII, the factor VIII-related antigen, and the ristocetin-related von Willebrand factor (vWF) and in stimulating a secondary rise in plasma factor VIII. Given either concentrate, the vWF activity, the antigen, and factor VIII levels were normalized in a patient with von Willebrand disease. However, correction of the prolonged bleeding time and control of bleeding occurred only with the cryoprecipitate. The bleeding-time corrective factor and the ristocetin-related vWF or platelet-aggregating factor are dissociable, distinct activites.


Polyene antibiotics. VII. Carbon-13 nuclear magnetic resonance evidence for cyclic hemiketals in the polyene antibiotics amphotericin B, nystatin A1, tetrin A, tetrin B, lucensomycin, and pimaricin1,2. Carbon magnetic resonance establishes conclusively that six polyene macrolide antibiotics containing keto groups (the heptaene amphotericin B, the tetraene-diene nystatin A1, and the tetraenes tetrin A, tetrin B, pimaricin, and lucensomycin) exist in the hemiketal form in solution. Their spectra all contain a hemiketal carbon's absorption near 97 ppm but lack a keto carbon's absorption near 210 ppm. The non-polyenic macrolide erythromycin, on the other hand, exists in the keto form.


Cytotoxic activity of purified guinea pig lymphotoxin against various cell lines. Cytotoxic activity of the purified guinea pig lymphotoxin (GLT) was tested on various blood cells and various cultured cell lines. GLT was generally ineffective on erythrocytes and lymphocytes, but it was somewhat effective on peritoneal exudate cells indicating GLT might play an important role in the regulatory mechanism of the cellular immunity. Considerable difference in target cell sensitivity was observed among the cultured cell lines tested. The sensitivity of L cells to GLT was significantly lowered by the trypsin treatment of the cells. Furthermore, porcine thyroglobulin glycopeptide exerted weak but definite inhibitory activity against GLT. These results suggest that the cytotoxicity of GLT might be triggered by the binding of GLT to the receptor sites of glycoprotein nature on the cell surface.


Carcinogenesis in tissue culture 27: heterotransplantation as an alternative in vivo assay for neoplastic state of cells in culture. Cells of serially cultured strains and newly-isolated cells originating from rat liver and lung were heterotransplanted into pouches of golden hamsters treated with antithymocyte serum (ATS). The "take" was determined by histopathological examination of nodules formed by 3 weeks after the transplantation. The results obtained under this criteria showed that 9 of 11 cell strains backtransplantable to rats and 2 of 4 strains untransplantable were in parallel with those in rats. Three kinds of uncultured liver cells resulted in "no take".


[Atypical retinopathia pigmentosa with central retinoschisis (Goldmann-Favre) (author's transl)]. A family is presented whose two daughters suffer from Goldmann-Favre disease. They have a retinoschisis and bone corpuscle like pigmentations. Both the girls have pathological ERG's (reduced b-waves, potentials up to 40 muV, no a-waves). The EOG of the younger is normal (Arden ratio 200%), but the EOG of the elder sister is pathological (Arden ratio right 120%, left 150%).


Cellular immune response in vitro: I. A requirement for time-dependent T-lymphocyte cell division of cytotoxic cells in the allogeneic response. Immunocompetent lymphoid cells cultured in vitro with allogeneic stimulator cells have been shown to produce T-lymphocyte populations which are specifically cytotoxic in vitro to the stimulatory cells whether normal or malignant. Although the culture requirements as well as the allogeneic requirements are known, the events leading to the production of T-lymphocyte cytotoxic cells is poorly understood. This study examines the role of cell division in the production of allogeneic cytotoxic T-cells in vitro. The elimination of cell division during the first 24 hr of allogeneic culture does not affect the cell-mediated cytotoxic immune response in vitro. Cell division is required, however, from 24 hr through 96 hr in culture and not necessary after 96 hr.


Histologic changes in saphenous vein aorta-coronary bypass grafts. The effect of the angle of the aortic anastomosis. Thirteen dogs were subjected to bypass grafting from the aorta to the left circumflex coronary artery with the saphenous vein to determine whether the angle of insertion of the saphenous vein into the aorta influences the functional and histologic fate of the grafts. The angle of the aortic anastomosis was obtuse in 6 dogs, acute in 5, and perpendicular in 2. Histologic examination of all 13 grafts 6 to 19 months (mean 9.4) postoperatively showed fibrous intimal proliferative lesions of variable severity along the entire length of the grafts, occasionally with extension into the native coronary arteries. Loss of medical smooth muscle and adventitial fibrosis also occurred in all 13 grafts. The extent and severity of these changes, however, were not related to the angle aortic anastomosis.