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Chediak et al. have reported that the titer of procoagulant antihemophilic factor (AHF:C; factor VIII:C) was significantly lower in obligate carriers of classic hemophilia who were daughters of affected men (paternal carriers) than in those whose fathers were normal by history (maternal carriers). In contrast, among 113 obligate carriers of hemophilia, no significant difference in procoagulant AHF titers was observed between paternal and maternal carriers. The concentration of AHF-like precipitating antigens, however, was significantly higher in maternal than in paternal carriers. This difference may have reflected in part the greater severity of disease in affected males in the families of maternal carriers.

A family is described in which Hageman trait appeared to be inherited in an autosomal dominant manner in contrast to previously reported families in which the disorder behaves as an autosomal recessive characteristic. Immunologic studies suggested that the molecular defect was similar to that of the autosomal recessive form and consisted of deficiency of antigens related to Hageman factor.

Calcium ions are known to accelerate the formation of a visible clot upon the addition of thrombin to plasma or fibrinogen. The experiments described demonstrate that this effect of calcium ions is not upon the rate of release of fibrinopeptides from fibrinogen, but rather upon the aggregation of soluble fibrin monomers or polymers, independent of the action of fibrin-stabilizing factor.

Sixty-nine cases of thrombocytopenia in which splenectomy had been performed have been reviewed. New cytological criteria are described for the diagnosis of disseminated lupus erythematosus by the examination of the splenic tissue. Six cases of thrombocytopenia associated with the ingestion of drugs and eight with infectious diseases responded promptly and permanently to splenectomy. The thrombocytopenia associated with disseminated lupus erythematosus (in 16 cases) and idiopathic thrombocytopenic purpura (in 39 cases) had a much more variable response. Approximately three-fifths of patients in each group had a remission sustained for at least 12 months following splenectomy. In general those patients who had thrombocytopenia for more than one year before surgery were less likely to respond to splenectomy.

THE interrelationship of psychological factors and organic disease has been frequently described. These interrelationships are particularly intriguing when the physical manifestation of disease appears to be a direct translation of some psychological pathology in the manner of a conversion reaction. This communication presents c;omocal data that suggest such a relationship between conversion reactions and the production of ecchymosis in patients with autoerythrocyte sensitization. This disease is a chronic purpuric state affecting adult and adolescent females, manifested by episodes of painful spontaneous bruising.1 The onset of the spontaneous bruise or ecchymosis is stereotyped. The patient's attention is drawn to a new lesion by a sudden pain described as sharp, stabbing, stinging, or burning. A lump then appears followed within a few minutes by an erythema. Within an hour or so an ecchymosis spreads from the periphery of the erythematous area. Although the erythema usually lasts no more than a

Ca++ potentiates the inactivation of human complement by streptokinase-activated plasmin and chloroform-activated bovine plasmin. The optimum concentration of Ca++ varies between 10–2 M and 10–3 M, decreasing with increasing concentrations of streptokinase. The susceptibility of the components of complement to inactivation by plasmin is different in the presence and absence of Ca++. C'2 and C'4 are most readily inactivated by plasmin in the presence of Ca++, while C'1 disappears first in the absence of Ca++. A large part of this C'1 disappearance is due to spontaneous inactivation. Similarities are pointed out between the influence of Ca++ on the inactivation of the components of complement by antigen-antibody systems and by plasmin.

Human complement is inactivated by plasmin, the proteolytic enzyme of plasma or serum active at or near neutrality. The addition of streptokinase to human serum, which converts plasminogen to plasmin, also causes the inactivation of complement components C'2 and C'4 and varying amounts of C'1. C'3 is the most resistant to inactivation by plasmin. Chloroform-activated human plasmin and bovine plasmin also destroy these components of complement, but are less effective than the streptokinase-activated enzyme. The inactivation of complement by the addition of streptokinase to human serum is inhibited by high hydrogen ion concentrations, low temperature, and elevated ionic strength. The inactivation of the components of complement in various fractions of serum is influenced by the available plasminogen and the content of plasmin inhibitors in these fractions. Certain similarities are pointed out between the components of complement and the factors in the plasmin system and between the inactivation of the components of complement by antigen-antibody reactions, by specific agents, and by plasmin. The possible significance of these relationships in immune hemolysis and complement fixation, and the possible role of the plasmin system in the instability of complement and the development of anticomplementary properties in serum are discussed.

Calcium ions accelerated the activation of the proteolytic enzyme of plasma from its precursor in the globulin fraction. Calcium did not appear to potentiate fibrinolysis by active bovine or human plasma proteolytic enzyme, nor did it accelerate the activation of the precursor of this enzyme by chloroform or by streptokinase. Experiments with partially purified plasma proteolytic enzyme suggested that the acceleratory effect of calcium was mediated indirectly. Since the concentration of calcium which was effective was comparable to that present in plasma, it is possible that the phenomena reported are of biological significance.

Through the kindness of Dr. J. H. Milstone we were able to perform clotting studies with purified bovine thrombokinase. Phenylmethyl sulfonylfluoride was found to inactivate bovine thrombin. Bovine thrombokinase, freed of traces of contaminating thrombin by treatment with phenylmethyl sulfonylfluoride, behaved in a manner similar to activated Stuart factor. Kinetic studies with this purified clotting factor and partially purified bovine proaccelerin supported the hypothesis that bovine proaccelerin is changed by the enzymatic action of thrombokinase to an agent which converts prothrombin to thrombin. The reaction between thrombokinase and bovine proaccelerin requires phospholipid and calcium and is blocked by soybean trypsin inhibitor. The similarities between human activated Stuart factor and bovine thrombokinase are apparent.

Modification of the kaolin clotting time has produced a reliable, simple assay system for antihemophilic factor. This system has been utilized for an investigation of the nature and mode of action of the circulating anticoagulant directed against antihemophilic factor. The anticoagulant has been shown to be present in equal amounts in plasma and serum, to be associated with fractions II and III of plasma prepared by the method of Cohn, to be relatively heat stable and to be stable for prolonged periods at -20 C. Investigation of the kinetics of the anticoagulant-antihemophilic reaction has demonstrated its temperature, time, pH, and substrate concentration dependency. The anticoagulant is not inactivated during the reaction with antihemophilic factor, and there is a proportional relationship between the anticoagulant and the antihemophilic factor during their interaction. No antigen-antibody manifestations could be detected during the anticoagulant-antihemophilic factor reaction. These characteristics support the hypothesis that the inactivation of antihemophilic factor by specific circulating anticoagulants is enzymatic.

A study was made of the family tree of two sisters with Hageman trait, an asymptomatic disorder of blood coagulation. In this family the defect behaves genetically as if it is due to the transmission of an autosomal recessive gene.

The proteolytic activity in chloroform-treated plasma euglobulins has been attributed to plasmin. Plasmin can digest both casein and fibrin. Epsilon aminocaproic acid, which inhibits the activation of plasminogen, the precursor of plasmin, by streptokinase, urokinase, and tissue activators enhanced the development of casein hydrolytic activity in a mixture of chloroform and plasma euglobulins. Fibrinolytic activity was also enhanced, but this was evident only if the epsilon aminocaproic acid was removed from the chloroform-treated euglobulins prior to assay. The reasons for the paradoxical enhancement of chloroform-induced casein hydrolysis by euglobulins containing epsilon aminocaproic acid are unclear. However, studies of optimal pH, heat stability, and the effect of ionic strength on the activation of the precursor of this proteolytic enzyme do not differentiate it from plasminogen.

The formation of C'1 esterase from C'1, the first component of complement, may be brought about by the action of plasmin or trypsin upon C'1s, a subcomponent of C'1. These enzymes also decrease the esterolytic activity of C'1 esterase. The formation of C'1 esterase was demonstrated by measuring the appearance of an agent or agents with esterolytic properties and the capacity to inactivate C'2 and C'4, attributes of C'1 esterase. The activity of the agent which evolved was blocked by serum inhibitor of C'1 esterase. The implications of these observations, that the formation of C'1 esterase during complement fixation is mediated by proteolytic processes, are under study. The possible inhibition of C'1q by soybean trypsin inhibitor is in agreement with this hypothesis.

Attempts have been made to elucidate the rôle of the endocrine glands in determining the susceptibility of animals to anaphylactic shock. Kepinow and Lanzenberg1found that thyroidectomy protected the guinea pig and rabbit against anaphylactic shock by inhibiting active sensitization with antigen. This, however, could not be repeated by Applemans2nor Yun;3and Fleisher and Wilhelmj4thought the protection was much less complete than the former authors believed. Blom5demonstrated the converse, that injection of thyroid preparations to sensitized guinea-pigs during the period of sensitization or shortly before the shocking dose, markedly increased the severity of anaphylactic shock; his method of injecting the shocking dose of antigen directly into the heart is, however, open to question.Adrenalectomy was shown by Kepinow,6in guinea pigs, and Flashman,7and Wyman,8in rats, to increase the susceptibility of these animals to anaphylactic shock. Epinephrin has long been known to protect against the symptoms of anaphylaxis.9Wolfram and Zwemer10showed that cortin administered from 2 to 6 hours before a shocking dose protected a large proportion of guinea pigs against anaphylaxis; and Dragstedt, Mills and Mead11noted that cortical extract administered to dogs 48 hours to 5 minutes before diminished the severity of, but did not prevent anaphylactic shock.Administration of parathyroid extract has led to contradictory results. Levinson and Matthews12claimed that amounts sufficient to raise the blood calcium of dogs to 15 mg % was without influence on the frequency of anaphylactic shock. Hajós,13on the other hand, found that the injection of parathyroid extract (Collip) decreased the sensitivity of guinea pigs to anaphylaxis.Hajós also reported that, while anterior pituitary extracts were without influence on anaphylaxis in the guinea pig, posterior lobe extract caused a decrease in sensitivity.

Activated Hageman factor, when injected into the rabbit ear chamber, produces a delayed and prolonged inflammatory response characterized by prominent sticking and emigration of leucocytes. In contrast, preformed bradykinin evokes an immediate and more transient response in which leucocytic emigration occurs less frequently. It is concluded that either Hageman factor produces its inflammatory effects by mechanisms other than kinin release, or bradykinin released endogenously has effects quite different from those resulting from a single injection of the exogenous material.

Purified preparations of the esterase derived from the first component of complement (C'1 esterase) increased vascular permeability in guinea pig skin, an effect inhibited by triprolidine, an antihistaminic agent, but not by soy bean trypsin inhibitor. The permeability-increasing and esterolytic properties of C'1 esterase were inhibited in parallel by the serum inhibitor of C'1 esterase, diisopropylphosphofluoridate and extremes of temperature and pH. Moreover, the permeability-increasing and esterolytic properties evolved in parallel when C'1 esterase was generated from its subcomponents. How C'1 esterase induces changes in vascular permeability remains unexplained, although the possibility that its action is mediated through a histamine-like agent is attractive.

It has been found that under a wide range of physico-chemical conditions a positive correlation exists between the rate of disappearance of hemolytically active, partially purified first component of human complement and the rate of activation of an esterase hydrolyzing N-acetyl-L-tyrosine ethyl ester. Both reactions follow the kinetic equation for second order autocatalysis, with an apparent energy of activation of 31,000 calories per mol. They occur optimally at pH 7.3–7.7 and are inhibited by ionic strengths greater than 0.15, by 5 x 105 M ethylenediaminetetraacetic acid, and by a heat-labile serum inhibitor which appears unrelated to any component of complement. The activation of first component to esterase resembles closely the activation of trypsinogen to trypsin. Partially purified first component, containing plasminogen, may also be activated to esterase by addition of streptokinase. The significance of these data with respect to the postulated existence of first component as a proesterase and its possible role in complement-"fixation" is discussed.

Studies on an esterase derived from partially purified preparations of the first component of complement are described. The esterase hydrolyzed certain synthetic amino acid esters, among which N-acetyl-L-tyrosine ethyl ester was most susceptible. This was hydrolyzed maximally between pH 7.5 and 8.2, and at 41°C. The esterase could not be identified with other previously described hydrolytic enzymes. An esterase with similar properties could also be eluted from antigen-antibody aggregates which had been treated with serum. Human serum contained a heat-labile inhibitor of the esterase which could not be identified with any of the known components of complement. The esterase was also inhibited by certain reducing agents. The experiments described support the early hypothesis that complement exerts its action enzymatically, but the physiological role of the esterase derived from preparations of complement is not yet clear.

The present report is an analysis of results obtained in the treatment of 124 patients with cirrhosis of the liver by means of a diet rich in protein and supplemented by concentrates of the vitamin B complex. In 1937 a preliminary statement1 was made concerning favorable therapeutic results obtained in 13 patients who had been observed during the course of one year. The improvement appeared to exceed chance expectations. A more comprehensive report2 in 1941 was made on 54 patients that had been studied over a four year period. The results tended to confirm the preliminary impression. With the new dietary regimen roughly one half of the patients showed signs of significant clinical improvement. These results were considerably better than those observed in a control series of 386 patients. However, since cirrhosis of the liver is a chronic disease and since its clinical course may vary considerably in

Autoerythrocyte sensitization is a chronic purpuric state, first described in 1955 by Gardner and Diamond.1 It is the purpose of this paper to demonstrate the frequency of emotional disturbances in patients with this disorder and to suggest that the character traits of these patients play a determining role in the syndrome itself.Autoerythrocyte sensitization is a disease of adult females, usually becoming clinically manifest after an injury, and characterized by repeated crops of ecchymoses, gastrointestinal bleeding, hematuria, abdominal pain, and neurologic abnormalities. Considerable disability may result from the pain of the cutaneous lesions and from hemarthroses. Gardner and Diamond suspected from the nature of this disorder that the patients had become sensitive to their own blood cells. They produced characteristic lesions by the intracutaneous injection of the patient's own blood or erythrocytic stroma.Since their initial report, only a few cases have been described.2-5 However in the last

Preparations of plasminogen, soluble in aqueous media, increased vascular permeability in guinea pig skin when activated by streptokinase or urokinase. The permeability-enhancing effect was inhibited by soybean trypsin inhibitor but not by triprolidine. Permeability-enhancing activity evolved when plasmin was incubated with fractions of plasma rich in prokinin. The experiments described suggest that plasmin exerts its permeability-increasing effect through the elaboration of kinins.

The fraction of human serum designated as C'1 esterase inhibitor is known to inhibit the action of C'1 esterase, a plasma kallikrein, and PF/Dil, an enzyme in plasma enhancing cutaneous vascular permeability. In the present study, C'1 esterase inhibitor has been found to block the actions of plasmin and the C'1r subcomponent of the first component of complement, and to retard the generation of PF/Dil. No inhibition of blood clotting or of the generation of plasmin was demonstrable.

Human C'1, a macromolecular complex composed of three subunits, is the zymogen for at least two distinct enzymes. Preparations of one subunit, C'1r, functioned as a protease which converted another subunit, C'1s, to C'1 esterase. The conversion of C'1s to C'1 esterase by C'1r was blocked by Liquoid, phenyl methylsulfonyl fluoride, and calcium ions, but not by soybean trypsin inhibitor, hirudin, or heparin. Preparations of C'1r also possessed two additional functions, i.e., the ability to hydrolyze certain synthetic amino acid esters and to participate in immune hemolysis. Evidence was presented which indicates that these three functions are properties of a single entity, C'1r, but not of the same portion of its molecular structure. These observations suggest that C'1r has at least two active sites, one for its reaction with C'1q, an additional subunit of C'1, and one for its reaction with C'1s; together, the three subcomponents, C'1q, C'1r, and C'1s, form a single functional unit, the first component of complement.

A fraction of globulin was prepared from human plasma which was deficient in prothrombin, thrombin, fibrinogen, plasma thromboplastin, and accelerator globulin. The preparation of globulin contained considerable potential proteolytic activity which could be activated by streptococcal fibrinolysin. This fraction of globulin accelerated the clotting of normal platelet-deficient plasma. However, the clot-accelerating effect of the globulin fraction was the same whether or not its proteolytic property had been activated. The addition of streptococcal fibrinolysin to normal platelet-deficient plasma did not accelerate coagulation. Nor did the addition of streptococcal fibrinolysin to hemophilic platelet-deficient plasma promote its coagulation. The data presented suggest that proteolysis by activated plasma proteolytic enzyme is not an essential stage in the coagulation of the blood.

Methods are described for the assay of chloroform-activated and fibrinolysin-activated plasma proteolytic enzyme, and for the determination of the inhibitory activity of plasma or other substances against chloroform-activated enzyme. The inhibitory activities of plasma, serum, and crude plasma albumin against chloroform-activated plasma proteolytic enzyme were proportional to the logarithm of the concentration of the inhibitory substance tested. This suggests that enzyme and inhibitor react in accordance with the law of mass action. The rate of fibrinolysis of recalcified plasma clots could not be related to the total proteolytic activity available in the plasma, nor to the inhibitory activity of fresh plasma or serum against plasma proteolytic enzyme. During the incubation of a recalcified plasma clot at 37°C., the inhibitory activity of its serum against plasma proteolytic enzyme decreased until a minimal stationary level was reached. The clot lysis time could be correlated directly with the time which elapsed until this minimal level was reached.

Local tetanus limited to one leg was studied in cats after intramuscular injection of tetanus toxin. 1. The electric and mechanical response of the affected muscle after a single stimulus to the intact sensory-motor nerve is greater in amplitude and duration than the response of the corresponding muscle of the unaffected leg (Fig. 1). 2. This augmented response of the muscle is associated with an augmented response arising from the ipsilateral portion of the spinal cord, while the contralateral part of the cord is unaffected, as demonstrated by electrographic records from the motor nerves (Figs. 2 to 5). 3. The augmented muscular response is abolished when the reflex arc is broken, but the augmented response in the spinal cord is independent of changes in the muscle, the neuromuscular junction, the afferent and efferent peripheral nerves, and the dorsal root ganglia. 4. The augmented spinal response develops in the absence of the peripheral signs of local tetanus. Hence the pathogenesis of the altered state in the spinal cord is independent of the peripheral effects of the toxin. 5. In local tetanus, therefore, the toxin injected intramuscularly acts selectively upon the segments of the spinal cord which supply the innervation of the injected area. 6. The augmented spinal response may be prevented by section of the nerve trunks supplying the area of injection prior to the injection of the toxin. 7. It is concluded that in local tetanus the toxin is carried to the spinal cord by way of peripheral nerves.