[Roentgenographic demonstration of the retrosternal lymph vessels. Animal experiments].

Roentgenographic demonstration of intraperitoneal organs and the retrosternal lymphatic system was obtained by insufflation of tantalum dust (particel size 1-5 mu) into the peritoneal cavity of 21 rats. No inflammatory reactions could be demonstrated. This contrastmedium remained over a considerably period of time in the peritoneal cavity and lymphnodes. This indicates the slow elimination of this metall.


[Measurement of renal blood flow with the Anger scintillation camera after selective intra-arterial injections of 133xenon and 99mtechnetium].

A "region of interest" technique for measuring renal blood flow is described, using as Anger camera and selective intra-arterial injection of 133Xe and 99mTc. The sequence of measurements has the advantage of providing information on regional blood flow. The values for RBF and RBFD obtained with 133Xe agree with those in the literature. Selective determination of the RCBF with the "region of interest" technique using 133Xe was not possible. Mean transit time for 99mTc was calculated after correction of the 99mTc indicator dilution curve with respect to recirculation and background activity. All typer of pathological changes in the kidney showed a statistically significant increase in mean transit time when compared with a control group. In patients with uncomplicated essential hypertension, the mean transit time was signigicantly reduced. A comparison of the mean transit time of 99mTc with the haemodynamic parameters derived from the 133Xe studies, and experiments with pharmocologically induced vasoconstriction indicates that the mean transit time represents a measure of the RCBF.


[Angiographic technique in assessment of renal mass lesions].

The advantages of pharmacoangiography and of selective renal venography as supplementory procedures to ordinary renal arteriography in the assessment of renal mass lesions are stressed. A new technique combining these procedures is introduced, and three examples are given.


[A new cannula for the introduction of catheters or other cannulas into the vascular system].

A new cannula is described consisting of a 25 cm. tube without a syringe attachment. It can be used for the introduction of other annuals (Hettler method) or for guide wires (Seldinger technique). The use of this cannula instead of the usual means of introducing catheters makes it possible to learn the technique quickly and to achieve correct placing of the cannula in the vascular lumen. The radiologist, using this method, achieves a safe and routine method very quickly. In this way angiographic examinations are made simpler they can be performed more quickly and the number of complications is reduced.


Drug eruption in a dog.

Scaly, slightly erythematous dermatitis developed in a young adult Vizsla dog after parenteral administration of acepromazine. The dog recovered after treatment with water baths, but the dermatitis recurred 4 months later, again after parenteral administration of the tranquilizer.


Metabolism of pyrimidine deoxyribonucleosides in Neurospora crassa.

The experiments in this report involve the following series of reactions which were previously demonstrated with purified enzyme preparations from Neurospora crassa: thymidine a yields thymine ribonucleoside b yields thymine c yields 5-hydroxymethyluracil d yields 5-formyluracil e yields uracil-5-carboxylic acid f yields uracil. The evidence for some of the reactions occurring in vivo has been incomplete and for others totally lacking. In this paper intact cells of Neurospora are shown to be capable of converting the substrates of each of the reactions to the corresponding products. Studies are described which were carried out in vivo and in vitro with the pyrimidineless strains pyr-4,uc-1,uc-2 and pyr-4,uc-1,uc-3, developed by Williams and Mitchell. The results reported in the present paper indicate that (reaction a) and the uc-3 mutation affects thymine 7-hydroxylase (reactions c,d, and e). Evidence is presented for the 2'-hydroxylase reaction being the major, if not only, way by which Neurospora can initiate the conversion of thymidine to the pyrimidines of nucleic acids and for the 2'-hydroxylation of thymidine and deoxyuridine being catalyzed by the same enzyme. Deoxycytidine was shown not to be hydroxylated in intact cells but instead deaminated to deoxyuridine, which in turn was converted to uridine. Further studies with the uc-3-carrying strain showed that an enzyme other than thymine 7-hydroxylase can also convert 5-formyluracil to uracil-5-carboxylic acid.


Generation of adenosine triphosphate in cytochrome-deficient mutants of Neurospora.

The fungus Neurospora crassa is known to possess a branched respiratory system consisting of the standard cytochrome chain and a cyanide-insensitive alternate oxidase. In the present experiments, the physiological function of the alternate oxidase has been analyzed by taking advantage of a number of cytochrome-deficient mutants, particularly poky f. Respiration, cellular ATP levels, and growth have been examined under the influence of three classes of inhibitors: inhibitors of the cytochrome chain (antimycin, cyanide), an inhibitor of the laternate oxidase (salicyl hydroxamic acid), and an uncoupling agent (carbonyl cyanide m-chlorophenylhydrazone). The results indicate that the over-all efficiency of the alternate oxidase in producing ATP and supporting growth is much less than that of the cytochrome chain. Depending upon the amount of oxidative phosphorylation at Sites II and III in the cytochrome chain, which varies from strain to strain, the efficiency of the alternate oxidase relative to that of the cytochrome chain ranges from 13% in wild type Neurospora to 18 to 21% in poky f, 35% in mi-3, and 57% in cyt-2. A comparison of the short term effects of cyanide and carbonyl cyanide m-chlorophenylhydrazone on cellular ATP in poky f suggests that, during respiration through the alternate oxidase, ATP can be produced both by substrate-level phosphorylation (accompanying glycolysis and the oxidation of alpha-ketoglutarate) and by oxidative phosphorylation at Site I. When cells are grown on sucrose, as much as 22% of ATP synthesis in the presence of cyanide occurs at Site I. When cells are grown on acetate to diminish the rate of glycolysis, the contribution of Site I becomes proportionately larger. Both the growth experiments and the short term inhibitor experiments reveal that ATP levels in Neurospora are kept  high be a feedback process which depresses ATP breakdown (and growth) very quckly after ATP synthesis is inhibited. Thus, poky f grows more slowly that wild type Neurospora and is inhibited still further when either the cytochrome chain or the alternate oxidase is blocked. Under all of these conditions, however, cellular ATP in poky f is maintained at a high level (about 3 mmol per kg of cell water, slightly above the values measured in the wild type strain). Continue.


Phosphorylation by inorganic phosphate of sodium plus potassium ion transport adenosine triphosphatase. Four reactive states.

Native solium and potassium adenosine triphosphatase from guinea pig kidney accepted a phosphate group from radioactive inorganic phosphate to form an acyl phosphate bond at the active site in the presence or absence of sodium ion. Magnesium ion was always required. In the presence of sodium ion and absence of adenosine triphosphate, there was no phosphorylation by inorganic phosphate. Addition of unlabeled adenosine triphosphate produced a potassium-sensitive phosphoenzyme which exchanged its phosphate-group with radioactive inorganic phosphate. The dephosphoenzyme was an intermediate in this exchange. The rate constant for dephosphorylation was about 0.05 per second. Addition of rubidium ion, a congener of potassium ion, to the potassium-sensitive phosphoenzyme produced a phosphoenzyme labeled from inorganic phosphate with a corresponding rate constant of 0.26 per s. This was a rubidium-complexed phosphoenzyme. Addition of magnesium ion to potassium-sensitive phosphoenzyme converted it into insensitive phosphoenzyme, the splitting of which was not accelerated by potassium ion or by adenosine diphosphate. Its rate constant was 0.07 per s. In the absence of sodium ion and adenosine triphosphate, inorganic phosphate was incorporated directly into a similar insensitive phosphoenzyme. In the presence of potassium ion or rubidium ion, inorganic phosphate was incorporated into a potassium-complexed or rubidium-complexed phosphoenzyme which exchanged 32-P with inorganic phosphate completely in less than 3 s. Incorporation of inorganic phosphate into a complex of the enzyme with the inhibitor, ouabain, is already described in the literature. Its rate constant was about 0.02 per s. Thus there appear to be at least four reactive states of the phosphoenzyme which equilibrate measurably with inorganic phosphate, namely, potassium-sensitive phosphoenzyme, potassium-complexed phosphoenzyme, insensitive phosphoenzyme, and ouabain phosphoenzyme. Two of these reactive states are functional intermediates in native sodium and potassium ion transport adenosine triphosphatase. The results are compatible with control of the reactivity of the active site by conformational changes in the surrounding active center and with regulation of the energy level of the phosphate group according to the kind of monovalent cation bound to the enzyme.


Sodium-potassium-activated adenosine triphosphatase of electrophorus electric organ. X. Immunochemical properties of the Lubrol-solubilized enzume and its constituent polypeptides.

Detergent (Lubrol WX)-solubilized sodium-potassium-activated adenosine triphosphatase ((Na+ + K+)-ATPase) of electrophorus electric organ contains two major constituent polypeptides with molecular weights of 96,000 and 58,000 which can be readily demonstrated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. These two polypeptides can be clearly separated and can be obtained in milligram quantities by preparative sodium dodecyl sulfate gel electrophoresis. The separated polypeptides, after removal of sodium dodecyl sulfate, and Lubrol-solubilized (Na+ + K+)-ATPase activity to some degree. Moreover, the degree of inhibition is directly proportional to the increasing amounts of antisera. The inhibition is maximal 4 weeks after the first injection. Immunodiffusion in 1% agar gel indicated that only Lubrol-solubilized enzyme antiserum, but not 58,000-dalton or 96,00-dalton polypeptide antiserum, gives one major precipitin band. However, specific complex formation between each polypeptide antiserum and Lubrol-solubilized enzyme occurs. This was demonstrated indirectly. After incubating Lubrol-solubilized enzyme with increasing amounts of polypeptide antisera at 37 degrees for 15 min, they were placed in the side wells of an immunodiffusion plate with antiserum against Lubrol-solubilized enzyme in the central well. The intensity of the precipitin band decreased with increasing amounts of polypeptide antisera. Thus, the results indicate that both 96,000-dalton and 58,000-dalton polypeptides are integral subunits of (Na+ + K+)-ATPase.


Partial resolution of the enzymes catalyzing photophosphorylation. XV. Approaches to the active site of coupling factor I.

1. Prolonged treatment of coupling factor I (CF1) from spinach chloroplasts with trypsin free of chymotrypsin yielded an active ATPase. The isolated preparation showed only two polypeptide chains (mol wt 55,000 to 60,000) on acrylamide gels run in the presence of sodium dodecyl sulfate. The three smaller subunits of CF1 were not detectable. The preparation no longer served as a coupling factor for photophosphorylation in either EDTA- or silicotungstate-treated chloroplasts. 2. An antiserum prepared against coupling factor I from chloroplasts inhibited the ATPase activity of the trypsin-treated CF1. In contrast, antisera prepared against the two individual (denatured) subunits did not inhibit the ATPase activity when tested either alone or together, although each interacted with the trypsin-treated protein, forming precipitin lines in Ouchterlony plates. 3. The trypsin-treated enzyme was still cold-labile, showing that the three smaller subunits are not required for this property. However, the enzyme was no longer sensitive to the natural inhibitor protein which is one of its subunits (subunit epislon), but was still sensitive to inhibition by the flavonoid quercetin. 4. Two equivalents of 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole were sufficient to inhibit about 80% of the ATPase activity of the coupling factor, irrespective of whether it contained two of five subunits. The inhibition was completely reversed by dithiothreitol. 5. Triated 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole was prepared. Treatment of the coupling factor with this tritium-labeled inhibitor followed by electrophoresis on acrylamide gels revealed that most of the radioactivity was incorporated into the beta subunit of the enzyme (molecular weight 56,000).


Isolation and characterization of Neurospora crassa plasma membranes.

The isolation and characterization of plasma membranes from a cell wall-less mutant of Neurospora crassa are described. The plasma membranes are stabilized against fragmentation and vesiculation by treatment of intact cells with concanavalin A just prior to lysis. After lysis, the concanavalin A-stabilized plasma membrane ghosts are isolated by low speed centrifugation techniques and the purified ghosts subsequently converted to vesicles by removal of the bulk of the concanavalin A. The yield of ghosts is about 50% whereas the yield of vesicles is about 20%. The isolated plasma membrane vesicles have a characteristically high sterol to phospholipid ratio, Mg2+-dependent ATPase activity and (Na+ plus K+)-stimulated Mg2+ATPase activity. Only traces of succinate dehydrogenase and 5'-nucleotidase are present in the plasma membrane preparations.


Comparative analyses of the kinetics and subunits of myosins from canine skeletal muscle and cardiac tissue.

Two types of canine cardiac myosins, myosin from the free wall of the right ventricle and the free wall of the left ventricle, were compared with canine skeletal muscle myosin from the gastrocnemius. The Vmax values for the ATPase reaction catalyzed by myosin were significantly different among the three types of tissues. For K+-activated myosin the Vmax values in micromoles of Pi per mg per min were: right ventricle, 0.57; left ventricle, 0.72; and gastrocnemius, 0.95. For Ca-2+ -activated myosin the Vmax values were: right ventricle, 0.32; left ventricle, 0.42; gastrocnemius, 0.50. All differences were significant (p smaller than 0.001). For all three types of tissues the Vmax values for NH4+ -activated myosin were the same (2.30). Light chains among all three types of tissues were immunologically identical, whereas the heavy chains of the two cardiac ventricles were immunologically identical with each other; however both were immunologically nonidentical with those of the gastrocnemius. The proportion of myosin light chains to heavy chains was different in the three types of tissue. Of the total protein present in each of the myosins, there was 18% in the light chains of right ventricle myosin, 10% in the light chains of left ventricle myosin, and 13% in the light chains of gastrocnemius. Both left ventricle myosin and myosin from gastrocnemius had significantly less C1d light chain, as compared to myosin from the right ventricle.


Acetyl phosphatidylethanolamine in the reconstitution of ion pumps.

Acetyl phosphatidylethanolamine was compared with phosphatidylethanolamine in the reconstitution of several biological membrane activities with the following results. 1. The proton pump reconstituted with the purple membrane of Halobacterium halobium and acetyl phosphatidylethanolamine was quite active. However, some differences in the kinetic properties, particularly in the decay rate, were noted between vesicles reconsituted with phosphatidylethanolamine and acetyl phosphatidylethanolamine. 2. Acetyl phosphatidylethanolamine could not replace phosphatidylethanolamine in the reconstitution of a Ca-2 plus pump with ATPase isolated from sacoplasmic reticulum. However, inclusion of suitable amounts of stearylamine or oleylamine during reconstitution yielded acetyl phosphatidylethanolamine vesicles with Ca-2 plus translocation activity comparable to that of phosphatidylethanolamine vesicles. 3. A mixture of acetyl phosphatidylethanolamine and stearylamine or oleylamine substituted for phosphatidylethanolamine in the reconstitution of mitochondrial hydrophobic proteins to form vesicles that catalyze 32-Pi-ATP exchange. Since phosphatidylcholine is also required in this system, these findings point to two functions of phosphatidylethanolamine, one related to the specific properties of its amino group, the other to a structural role of its small polar head group. A hydrophobic alkylamine can fullfill the first function, acetyl phosphatidylethanolamine the second. 4. The importance of the charge was also observed in experiments with the reconstituted rutamycin-sensitive ATPase of mitochondria. After depletion of phospholipids from the hydrophobic proteins, ATPase activity and rutamycin sensitivity were restored only if a phospholipid as well as the appropriate charge were present.


The red cell membrane contains three different adenosine triphophatases.

Phosphorylated intermediates in the hydrolysis of ATP by human red cell membrane adenosine triphosphatases have been detected using [gamma-32-P]ATP. Intermediates formed in the presence of Mg2+ alone (MG-2+-ATPase), Mg-2+ and Na+ ((Na+,K+)-ATPase), and Mg-2+ and Ca-2+ (Ca-2+-ATPase) were separated by polyacrylamide gel electrophoresis in sodium dodecyl sulfate at pH 2.4, indicating that these three ATPases are different molecular species. There are roughly 100, 150, and 400 copies per cell, respectively, of the three ATPases.


Amyloid in the aged heart: frequency and clinical significance.

Cardiac amyloid accumulation is a common autopsy finding in elderly persons, the frequency increasing with each advancing age decade. In most cases the deposits are microscopic, confined to the atrial subendocardium, and of dubious significance. In a few cases, however, there is much more extensive cardiac involvement, with infiltration of the ventricular musculature and intracardiac conduction system. In the authors' recent autopsy study of 100 patients over 60 years of age, 10 had diffuse cardiac amyloidosis with biventricular involvement. Microscopically, the amyloid deposits often resembled foci of interstitial fibrosis. Significant cardiac enlargement (weight greater than 380 gm) was present in only 2 of these patients, and in only one of them were the clinical signs and symptoms clearly attributable attributable to the amyloid infiltration. It was difficult to assess the possible contribution of ventricular amyloid to congestive failure since most of the patients also had coronary arteriosclerosis. In the aged, clinically significant cardiac amyloidosis associated with cardiac enlargement appears to be part of a disease spectrum more commonly characterized by focal, clinically insignificant cardiac amyloid deposits. Yet several features suggest that senile cardiac amyloidosis may differ from the clinically significant cardiac amyloidosis occassionally seen in younger patients.


Ir-associated murine alloantigens: demonstration of multiple Ia specificities in H-2 alloantisera after selective absorptions.

Quantitative absorptions of anti-H-2 alloantisera with increasing concentrations of T cell-derived tumor cells of donor H-2 type has been found to remove antibodies to H-2K region antigens selectivelymsimilar results were obtained after absorptions with thymocytes of the donor strainl In each case residual antibodies reacting with Ir-associated B cell alloantigens (Ia antigens) were unmasked by this procedure. Antisera which initially contained comparable titers of Ia and H-2 activity were depleted of H-2 activity at absorbing cell concentrations which did not appreciably alter the Ia reactivities, indicating that Ia antigens, if expressed on these T cell populations, must have been present at very low concentrations relative to H-2D and H-2K region antigens. Further absorptions with spleen cells from cross-reactive strains demonstrated a multiplicity of shared and unique Ia specificites in these sera. The multiple Ia antigens detected appeared all to be codominantly expressed on the same subpopulation of lymphoid cells.


Macrophage cytotoxicity in the mouse immune response against a skin allograft.

Macrophage-rich peritoneal cell populations from C57BL/6 mice grafted with DBA/2 skin were found to be cytotoxic against 51Cr-labeled target cells from the donor strain. Normal peritoneal macrophages were also rendered cytotoxic by incubation with acellular supernatants of mixed lymphocyte cultures (MLC) between an allograft recipient and a donor mouse. Supernatants alone were not cytotoxic. The macrophage arming factor(s) was found in supernatants when the MLC was performed after more than 6 to 9 days following grafting. In order to produce MAF, sensitized lymphocytes must usually be stimulated in a specific way by donor type cells. The armed macrophage cytotoxicity was, however, not found to be specific in these experiments.


Inhibition of proliferation of lymphoma cells and T lymphocytes by suppressor cells from spleens of tumor-bearing mice.

We have recently demonstrated suppressor cells in spleens of mice bearing tumors induced by Moloney murine sarcoma virus (MSV) which were non-T cells and inhibited phytohemagglutinin Molney (PHA)-induced DNA synthesis of syngeneic normal spleen cells. From the present study, the suppressor cells appeared to be macrophages since they were radioresistant, inactivated by carrageenan, and removed by adherence columns and an iron/magnet technique. We have also found that suppressor cells were still fully active when added 16 hr after the mitogen, thus indicating that early mitogen-induced changes were not the target of suppressive action. It appeared that suppressor cells inhibited metabolic events related to the initiation of DNA synthesis and that they had a selective effect on proliferation-dependent lymphocyte effector functions. PHA-induced cytotoxic reactivity which in our system is largely independent of DNA synthesis was not depressed but actually enhanced in MSV spleens. Cytotoxicity of MSV spleen cells against syngeneic lymphoma cells was unaffected by suppressor cells whereas lymphocytes stimulation by mitomycin C-treated syngeneic lymphoma cellls was inhibited. MSV spleen cells also inhibited DNA synthesis of cultured murine lymphoma cells. This function was only slightly diminished after treatment with anti-omicron serum plus guinea pig complement. Furthermore, spleen cells from MSV tumor-bearing nude mice were as effective as spleen cells from their heterozygous littermates, thus suggesting that T lymphocytes are not the main effector cells of inhibition of lymphoma cell DNA synthesis. The inhibitor cells were radioresistnant, inactivated by carrageenan, and removed by adherence columns and the iron/magnet technique. These data strongly suggest that the inihibitor cells of lymphoma cell DNA synthesis are macrophages and that they belong to the same group of cells as the suppressor cells of PHA-induced lymphocyte proliferation.


The suppressive effect of immunization on the proliferative responses of rat T cells in vitro.

DNA synthesis (3H-thymidine incorporation) of DA rat spleen and lymph node cells cultured with PHA or (DA x Lewis)F1 lymph node cells was profoundly inhibited by pretreatment with large i.v. doses of antigen (greater than 100 mg OA or greater than 10(10) SRBC) 24 hr before culture. This effect, seen as early as 3 to 6 hr after antigen administration, had disappeared within 7 days. Similar depressed spleen cell responses were seen 9 days after footpad injection of 200 mug OA in complete adjuvant. Lymph node responses showed a similar inhibition only in the presence of specific antigen in culture. In all experiments in which antigen administration resulted in decreased responses, removal of glass wool adherent cells restored responsiveness to normal. The failure of purified macrophages to reestablish inhibition favored the interpretation that an adherent suppressor cell distinct from the macrophage is activated by antigen in vivo to regulate T cell responses in vitro.


T mitogens trigger LPS responsiveness in mouse thymus cells.

Mouse thymus cells are essentially unresponsive to lipopolysaccharide (LPS) stimulation. However, when cultured with minimally mitogenic levels of concanavalin A or submitogenic ratios of mitomycin-treated allogeneic spleen cells, in combination with LPS, they demonstrate levels of DNA synthesis de novo or greater than those induced by the T cell mitogen alone. Dose-response kinetics were characteristic of LPS. The subpopulation containing the LPS responsive cells was of low net buoyant density. Neither phytohemagglutinin nor pokeweed mitogen acted synergistically with LPS in this model to trigger thymus cells. The data suggest that LPS triggering may involve interaction with a T cell subpopulation.


Biological expressions of lymphocyte activation. IV. Concanavalin A-activated suppressor cells in mouse mixed lymphocyte reactions.

Thymus-derived lymphocytes (T cells) from mouse spleen, activated in vitro or in vivo with concanavalin A (Con A), suppress proliferative responses of syngenic lymphocytes in mixed lymphocyte reactions (MLR). Replication in vitro was not required for expression of suppressor activity by Con A-activated cells and was blocked in MLR by treating suppressor cells with mitomycin C or irradiation. Kinetics of MLR responses and viability of cultures were not altered by addition of activated suppressor cells. The data are consistent with a direct inhibitory effect of suppressor T cells on antigen-induced DNA replication. These observations extend a model previously described for regulation of antibody synthesis by Con A-activated T cells to control of cell-mediated immune responses. This model should be particularly useful in further definition of regulatory T cell subpopulations, and in investigation of interactions and relationships between such populations.


Studies on the synthetic capacity and antigenic expression of glutaraldehyde-fixed target cells.

Mastocytoma cells (P815 of the DBA/2 strain) treated with increasing concentrations of glutaraldehyde were concurrently evaluated for their ability to incorporate exogeneous uridine, thymidine, and amino acids. The antigenic expression and membrane integrity of these treated cells were assayed by measuring their susceptibility to lysis by antibody and complement and by T-effector cells. The concentrations of glutarladehyde required to effect target cell antigen display were greater than those required to inhibit totally the cell's protein and nucleic acid synthetic processes. Thus, P815 cells treated with 0.15% glutaraldehyde for 10 sec were unable to incorporate either amino acids or nucleotides into macromolecules, but were readily lysed by effector T cell populations, and by alloantibody in the presence of complement. Target cells treated with glutaraldehyde concentrations in excess of 0.25% for 10 sec were resistant to both forms of immune lysis. In keeping with these observations, monolayers of P815 cells treated for 10 sec with 0.15% glutaraldehyde, were capable of specifically depleting T-effector cells from a cytolytically-active spleen cell population. After treatment with higher concentrations of glutaraldehyde (0.3%), however, the monolayers lost their capacity to adsorb effector cells. Although P815 cells treated with glutaraldehyde continued to exhibit H-2d alloantigen, neither these cells nor glutaraldehyde-treated DBA/2 spleen cells induced significant blastogenesis or stimulated the production of cytolytically active effector cells in mixed leukocyte cultures.


Migration inhibitory activity in serum and cell supernatants in patients with Sezary syndrome.

The abnormal lymphocytes from patients with the Sezary syndrome produce macrophage migration inhibitory factors (MIF) both in vitro and in vivo. Five of six individuals studied had significant serum MIF activity and one had borderline activity. In contrast, in this study 47 normal individuals and 9 of 10 patients with extensive skin disease other than that of the Sezary syndrome had no such activity. Since the neoplastic T cells in patients with the Sezary syndrome localize in the skin, their production of chemical mediators in that site may be responsible for the generalized exfoliative erythroderma seen in that disease.


Adult murine lymph node cells respond blastogenically to a new differentiation antigen on isologous and autologous B lymphocytes.

Cells derived from lymph nodes (LN) of adult CBA mice respond blastogenically to mitomycin-treated autologous, as well as isologous spleen cells. This isogeneic LN-to-spleen (mixed lymphocyte culture) is best obtained when both responder and stimulator cells are derived from donors greater than 10 weeks of age. Responsive cells appear restricted to LN since they could not be detected in adult spleen, marrow, or thymus. LN cells do not require the presence of spleen in order to differentiate into responder cells since those derived from neonatally splenectomized mice are fully active. Stimulator cells appear in the spleen, bear Ig on their surfaces, and can be detected in spleens of irradiated, bone marrow-reconstituted mice. Experiments comparing the responsiveness of adult LN cells and that of neonatal T cells toward mitomycin C-treated lymphoid cells from a variety of sources suggest the presence of two iso-antigens on B lymphocytes. Since both antigens apparently are absent on precursor bone marrow cells and develop with time, they have been classified as murine differentiation antigens 1 and 2 (MDA-1, MDA-2). Whereas both appear in the spleen, only one, MDA-1, is also detectable by this methodology in LN. Both MDA-1 and MDA-2 activate neonatal T cells, but MDA-2 triggers only adult LN. Whereas MDA-2 developed in an x-irradiated, bone marrow-reconstituted spleen, MDA-1 did not over a 9-week interval.


Immunologic functions of isolated human lymphocyte subpopulations. III. Specific allogeneic lympholysis mediated by human T cells alone.

The studies presented herein have evaluated both the specificity and cellular basis of cell-mediated lympholysis (CML) in man. An efficient and quantitative 51Cr release assay was utilized to study the role of highly purified human T and B cells in CML. After in vitro sensitization human T cells develop the capacity to kill specifically allogeneic cells to which they were sensitized. In contrast, B cells were neither triggered to proliferate nor activated to kill allogeneic targets. B cells were not activated to kill even when sensitized in the presence of potentially "helper" T cells, nor did they block T cells from killing during the effector phase. Cell-free supernatants taken from active in vitro sensitization cultures were not lympholytic and did not modulate T cell killing. Hence, these studies show that both the afferent and efferent phases of human CML are T cell functions.


A scanning and transmission electron microscope study of the luminal coating on Dacron prostheses in the canine thoracic aorta.

The sequence of events occurring in a woven Dacron bypass in the canine thoracic aorta was followed from the first exposure to flowing blood for a period of 27 months. After 10 and 40 minutes exposure to flowing blood the surface was covered with masses of leukocytes, platelets, red cells, and some fibrin. By 24 and 48 hours most of the leukocytes had disappeared leaving the two end areas of the bypass covered with a thick film composed of red cells entrapped in a fibrin net. In the central portion of the bypass the Dacron fibers were covered by a rough film of particulate material and widely spaced clumps of platelets and occasional leukocytes connected by a few fibrin strands. AT 6 AND 7 DAYS THE SURFACE COVERING CONSISTED PRIMARILY OF RED CELLS AND FIBRIN. Between 2 and 8 weeks this was replaced by a matrix that was composed mostly of cells resembling fibroblasts or transition forms between these and smooth muscle cells. Some typical smooth muscle cells were observed in grafts 4 months or older. A sheet of flat cells had grown a few millimeters below the anastomoses at 7, 12, and 14 months. By 27 months the entire length of the graft had a patchy covering of endothelium. Only occasional isolated platelets were stuck to the exposed collagen. Thickening of the intraprosthetic lining was associated with infiltration of the cell-fiber matrix by masses of red cells. Thus it appears that under proper conditions of flow, thrombogenic surfaces such as Dacron and collagen are "conditioned" to become nonthrombogenic by the limited deposition of blood elements.


Potency of fragmented IgG: two studies of postexposure prophylaxis in type A hepatitis.

To determine the effect of fragmentation on potency of immune globulin preparations, two comparisons were carried out. In one study, the immune globulin was derived from American plasma; in the other, the source was Israeli plasma. In each of the two studies, three materials were given to household contacts of icteric hepatitis: (1) human albumin as a placebo; (2) immune globulin with the IgG intact; and (3) immune globulin of the same lot with the IgG deliberately fragmented by added fibrinolysin. Comparable reductions in secondary attack rates were achieved with fragmented and unfragmented materials from both lots. Fragmentation, therefore, had no deleterious effect. In addition, it was found that American globulin is comparable to Israeli globulin for protection against strains of Type A hepatitis prevalent in Israel. Administration in the second half (last 15 days) of the incubation period did not reduce the frequency of icteric disease.


Evidence for a B-cell -like helper function in mixed lymphocyte culture between immunocompetent thymus cells.

The immunocompetent subpopulation by mouse thymus cell (TH-2) was isolated by buoyant density centrifugation and by hydrocortisone pretreatment. TH-2 cells undergo a proliferative one-way or two-way mixed lymphocyte culture (MLC) response only when cultured with allogeneic or congenic peripheral lymphoid cells. However, mixtures of allogeneic TH-2 cells alone do not proliferate in either one-way or two-way MLC reactions. Such MLC mixtures are proliferative only if mitomycin-blocked peripheral lymphoid cells are also present in the mixture. The peripheral helper cell has been found to be of low net density, non-adherent, insensitive to anti-thy-1 serum cytotoxicity, but sensitive to the cytotoxic effets of anti-immunoglobulin serume plus complement. The helper effect does not depend on proliferation nor does it appear to involve demonstrable soluble mediators. The nature of failure of MLC between TH-2 subpopulations appears to be dependent on the exppression of some product of the K, I regions of the H-2 locus. Possible mechanisms by which a B-cell-like helper cell triggers TH-2 proliferation are discussed terms of the present knowledge of specific alloantigen receptor on T and B cells, and the immunoglobulin Fc region receptors on T cells.


H-2 compatibility is required for T-cell-mediated lysis of target cells infected with lymphocytic choriomeningitis virus.

Maximal cell-mediated lysis of targets infected with lymphocytic choriomeningitis virus occurs only within a H-2 compatible system. Syngeneic immune spleen cells are at least 100 times as effective as are allogeneic lymphocytes. Reciprocal restriction of cytotoxic T-cell activity has been shown to operative between H-2k, H-2d, and H-2b. Experiments with cogenic mice have localized the effect to the H-2 gene complex. Furthermore, the observation that lymphocytes from H-2a mice cause high specific 51Cr release from either H-2d virus-infected cells, indicates that identity at either the K or the D end of the H-2 gene complex is sufficient for this lytic interaction.


The surface morphology of human B lymphocytes as revealed by immunoelectron microscopy.

Surface immunoglobulins (sIg) were detected on human lymphocytes by immunoelectron microscopy with peroxidase-conjugated antibodies. Blood, marrow, and thymus cells from normal individuals and patients with lymphoproliferative disorders were examined. Samples were fixed before exposure to specific reagents. Normal lymphocyts with detectable sIg, i.e. B lymphocytes, were characterized by a villous surface; nonlabeled blood lymphocytes and thymocytes were smooth cells. Intermediate cells were also found which in sections appeared moderately villous and labeled, thus identified as B lymphocytes. Further evidence for a relationship between villous surface and sIg was given by the finding of a few lymphocytes with polar concentration of labeled microvilli. In chronic lymphocytic leukemia patients, most cells exhibited a villous surface with parallel variations of the number of microvilli and of anti-immunoglobulin-binding capacity. However, some labeled smooth blastic cells were also observed. On the other hand, abnormal lymphocytes from Sézary's syndrome which could exhibit segments of villous membrane had no detectable sIg. This study confirms that in most cases human B lymphocytes have a characteristic surface appearance and that the detection of sIg in normal lymphocytes correlates with the presence of microvilli.


Sequential responses of mouse spleen T cells in mixed lymphocyte culture-induced cytolysis.

T cells triggered to blast transformation and proliferation by histoincompatible cells have the capacity of reverting "back" to lymphocytes. These "secondary" lymphocytes and their progeny cells are able to respond repeatedly to the same allogeneic stimulus in vitro.


Measurement of intracardiac pressure through a valve-tipped catheter. Technical note.

Intracardiac pressure recording is a simple and effective way of obtaining optimal placement of the cardiac catheter during shunting procedures. We have also found this method applicable when using a catheter with a slit in its tip. Pressure waves can be accurately recorded by means of a constant rate liquid flow through the catheter.


The critically ill premature infant with patent ductus arteriosus and pulmonary disease-an echocardiographic assessment.

The effect of left-right shunting upon left ventricular and atrial dimensions was assessed by means of 129 serial echocardiographic studies in 37 premature infants with patent ductus arterisus. The cardiac dimensions of asymptomatic premature infants and those with pulmonary disease were similar and served as control subjects. Left cardiac dimensions were enlarged in infants with significant PDA. Dimensions were greatest in surgically treated infants (post-operatively they returned rapidly to normal) in contrast with the persistent enlargement observed in some medically treated infants. Serial echocardiography was a valuable adjunct to clinical assessment of the course of PDA in premature infants with pulmonary disease.


Relationship between fibrinolysis of cultured cells and malignancy.

Cells cultured from various human and nonhuman malignant and normal tissues as well as mammalian cells transformed in vitro were examined for their ability to induce fibrinolysis. Generally, except for normal cells derived from lung or kidney, malignant cells had a greater ability to induce fibrinolysis than did their normal counterparts. A correlation existed between the abilities of the cells to induce fibrinolysis, grow in soft agar, and form tumors in immunosuppressed hosts.


Characterization of a bacteriophage T4 mutant lacking DNA-dependent ATPase.

A DNA-dependent ATPase has previously been purified from bacteriophage T4-infected Escherichia coli. A mutant phage strain lacking this enzyme has been isolated and characterized. Although the mutant strain produced no detectable DNA-dependent ATPase, growth properties were not affected. Burst sizes were similar for the mutant phage and T4D in polA1, recB, recC, uvrA, uvrB, uvrC, and various DNA-negative E. coli. UV sensitivity and genetic recombination were normal in a variety of E. coli hosts. Mapping data indicate that the genetic locus controlling the mutant occurs near gene 56. The nonessential nature of this gene is discussed.


Vaccinia virus infection of HeLa cells. I. Synthesis of vaccinia DNA in host cell nuclei.

The replication of vaccinia virus is thought to take place exclusively in the cytoplasm of host cells. However, using DNA-DNA hybridization techniques, it can be shown that a significant fraction of the synthesis of vaccinia DNA takes place in the nucleus as well as the cytoplasm. The (3H) thymiding pulse-labeled vaccinia DNA synthesized in the nucleus reaches a maximum at about 3 h after infection, corresponding to the time of maximal DNA synthesis in infected cells. At this time host DNA synthesis drops to about 25% of the rate of the uninfected cells. Even with short labeling times (2 min) the nucleus is found to contain 60% of the incorporated (3H)thymidine, much of which is in vaccinia DNA. Prior inhibition of host nuclear DNA synthesis with mitomycin C, followed by removal of the antibiotic causes a subsequent inhibition of vaccinia DNA synthesis and complete suppression of mature virus. Purified nuclei, isolated from vaccinia-infected cells, also synthesize vaccinia DNA in vitro. Over 90% of the DNA synthesized in vitro by isolated nuclei contain vaccinia-specific sequences.


Elution and uncoating of Coxsackievirus B3 by isolated HeLa cell plasma membranes.

Plasma membranes isolated from HeLa cells on discontinuous sucrose gradients were assayed for their capacity to elute and uncoat coxsackievirus B3 at 37 C. Because the viral receptors are limited to the surface of HeLa cells, the addition of radioactively labeled virus to the cells prior to cell homogenization provided a useful marker for locating the plasma membranes during the fractionation procedure. Four bands were formed on the discontinuous sucrose gradients with approximately 70% or more of the membrane-associated viral label being recovered in the most dense bands, designated as bands 3 and 4. Bands 3 and 4 also possessed the plasma membrane marker enzymes, Na+, K+ adenosine triphosphatase and 5'-nucleotidase and revealed typical structures characteristic of plasma membranes as revealed by electron microscopy. Pelleted and washed membranes from band 3 both eluted and uncoated B3 32P-labeled virus, whereas membranes from band 4 eluted virus but failed to uncoat it. The membranes from band 4 were shown to inhibit the viral uncoating activity when mixed with membranes of band 3. Characteristically, unfractionated homogenates of cell membranes eluted but did not uncoat virus. The finding of a naturally occurring inhibitor of virus uncoating provides for the first time a way to distinguish between the membrane activities of virus elution and virus uncoating. The inhibitor remains to be characterized.


Prevalence of Dictyocaulus viviparus infection in Rocky Mountain elk in Teton County, Wyoming.

Dictyocaulus viviparus infections in Rocky Mountain elk (Cervus canadensis of Teton Countywere surveyed by fecal analyses during spring, summer and winter  and by fecal analyses and necropsies during fall hunting seasons, 1968-1973. Prevalance of the lungworms was relatively high: 32-70% during the spring; slightly lower, 30-47%, during the summer; 21-39% in the fall; and declined to the annual low of 8-19% during the winter. Conversely, elk summering on Big Game Ridge showed an increase in prevalance of D. viviparus from 1969 to 1973. Decreases in prevalance of lungworms were noted on the National Elk Refuge at Jackson after management changes were effected in 1971.


Chymopapain chemonucleolysis in lumbar disk disease.

Experience with chymopapain chemonucleolysis in 1,200 cases of lumbar disk disease indicates that it is an effective method of relieving pain resulting from an abnormal intervertebral disk. Complications were few; anaphylaxis occurring immediately after injection was by far the most serious. With the use of a corticosteroid and an antihistamine in preoperative preparation, the incidence of anaphylaxis had dropped, although it still occurs. On long-term follow-up, chemonucleolysis appears to be as good as laminectomy in properly selected cases.


Blood flow velocity in the carotid artery as a measure of myocardial contractility.

An attempt was made to derive a useful noninvasive index to evalute a change in myocardial contractile state using transcutaneous Doppler flow-velocity curve from the carotid artery. In 5 mongrel dogs and in 43 patients with various heart disease, Doppler flow velocity curves were obtained from the ascending aorta intravascularly using a Doppler catheter and/or from carotid artery transcutaneously using a Doppler probe. The first derivative of left ventricular pressure (dp/dt) and electrocardiogram (ECG) were recorded simultaneously. The following 3 indices were measured from the Doppler flow-velocity curves: (1) macimum acceleration of blood flow (dv/dt), (2) time from onset of ejection to peak flow (time-to-peak), (3) time interval between the beginning of Q wave of ECG to the peak of Doppler flow velocity curve (ECG Q-Doppler peak). Among these 3 indices, only ECG Q-Doppler peak demonstrated a significant correlation between the values measured intravascularly and transcutaneously. Also, only ECG Q-Doppler peak showed significant correlation with maximum of dp/dt (max dp/dt). Since ECG Q-Doppler peak showed correlation with heart rate, the difference between observed and predicted ECG Q-Doppler peak (delta ECG Q-Doppler peak) was calculated to exclude the effect of heart rate. Predicted value of ECG Q-Doppler peak was calculated from the regression equation between heart rate and ECG Q-Doppler peak in the separate experiments. There was significant correlation between delta ECG Q-Doppler peak and max dp/dt. In 15 patients with coronary artery disease and in 16 healthy subjects, delta ECG Q-Doppler peak and the other noninvasive method (systolic time intervals) were measured. Delta ECG Q-Doppler peak showed better result in the separation of 2 groups than by systolic timeintervals. It was concluded that delta ECG Q-Doppler peak is a useful index to evaluate the myocardial contractile state since this index is readily obtained noninvasively.


Relief of congenital obstruction to left ventricular outflow with a ventricular-arotic prosthesis.

Hypoplasia of the aortic valve annulus and ascending aorta is a rare form of congenital aortic stenosis, with a poor prognosis. Replacement of the aortic valve and ascending aorta with a suitable prosthesis is feasible, but the hypoplastic valve annulus must also be enlarged if an adult-sized aortic valve is to be placed in the subcornary position. In an effort to develop a new method of surgical treatment for this congenital abnormality, we fabricated a prosthesis which can be interposed between the left ventricular apex and descending thoracic aorta. This prosthesis is coupled to a 25 mm. Dacron graft and xenograft valve and is lined with flocked Dacron fibrils to encourage formation of a stable biologic lining and prevent thrombus buildup. The prosthesis was implanted in a 22 year old male patient of the Children's Hospital Medical Center without difficulty. Postoperative cardiac catheterization idicated that all of the contrast material was ejected from the left ventricle through the prosthesis. The patient was discharged from the hospital taking sodium warfarin and remains entirely asymptomatic.


Lymphocyte transformation test in fixed drug eruption.

Twenty-one patients with fixed drug eruption were studied with the lymphocyte transformation test. In no patient was there blast transformation when the responsible drug (1:10,000 dilution) was added to the lymphocyte culture. The addition to the lymphocyte culture of 0.4 ml of autologous serum, taken at the acme of the clinical reaction, produced blast transformation of the lymphocytes, and when the responsible drug (1:10,000 dilution) was added to this system, blast transformation increased by several times. The addition of the responsible drug to autologous serum obtained during clinical remission produced a minimal or negative response. These findings suggest that during fixed drug eruptions a blast transforming factor appears temporarily in the serum and increases its activity in the presence of the responsible drug. This factor spontaneously diminishes or disappears a few days after clinical exacerbation.


A disorder of muscle lipid metabolism and myoglobinuria. Absence of carnitine palmityl transferase.

Two brothers, 29 and 33 years of age, had recurrent myoglobinuria, renal failure and azotemia, but were otherwise normal, without apparent muscle weakness or exercise intolerance. Ischemic exercise resulted in normal lactate production. Muscle glycogen content and activities of phosphorylase and phosphofructokinase were normal. Plasma triglycerides were elevated (500 mg per deciliter) on a regular diet and rose during fasting. During a 72-hour fast, serum creatine phosphokinase rose more than 10 times, and myoglobin was detected in urine. Plasma ketone production was minimal during fasting, but prompt ketonemia ( a normal response) occurred after ingestion of medium-chain triglycerides. Carnitine palmityl transferase activity was virtually absent in crude muscle extracts and mitochondrial fractions. Lack of this enzyme impairs long-chain fatty acid utilization, reflected in increased content of plasma free fatty acids and plasma triglycerides. Depletion of ATP because of this metabolic block in muscle may account for the attacks of myoglobinuria.


Hereditary carnitine deficiency of muscle.

An eight-year-old boy with slowly progressive muscle weakness was found to have Oil red O positive vacuoles in predominanty type i muscle fibers. Subsequent studies demonstrated markedly reduced skeletal muscle carnitine (0.24 mumoles per gram; normal 1.64 to 3.34). Serum carnitine was normal. Although both parents were clinically normal, muscle carnitine levels were low in both (mother 0.60; father 0.90 mumoles). There was no clinical evidence of cardiac disease but the patient had ventricular hypertrophy by electrocardiography, vectorcardiography, and echocardiography. Treatment with prednisone resulted in clinical improvement but no change in muscle histology. Our studies suggest that the carnitine deficiency of muscle in this case may be due to impaired carnitine entry into muscle and that this form of disease can be inherited as an autosomal recessive disorder.


[The phenomenon of water entrapment in the formation and course of edema. A physiopathological mechanism useful for therapeutic purposes].

Results with a diffusor enzyme (thiomucasi) in the medical management of oedema in 29 subjects are presented. This substance was used on the assumption that interstitial mucopolysaccharides trap water by means of a polymerisation-depolymerisation process, so that its release necessitates rupture of chemicophysical bonds. The results point to the effectiveness of the drug and the soundness of this interpretation.


[The value of laparoscopy in the study of portal hypertension].

Up to now, laparoscopy has always been considered as only marginally importan in the study of portal hypertension whereas in fact it should become one of the fundamental examinations in this field. The technique not only contributes, by direct liver exploration, to a more precise judgment regarding the underlying disease, with the possibility of earlier recognition of pre-hepatic, intra-hepatic and post-hepatic forms of portal hypertension, but also makes it possible to detect the earliest signs of hypertension. Further, and most important, laparoscopy extends investigation of the collateral circulation to the sector of the small peritoneal vessels which are inaccessible with other techniques, demonstrating alterations are of decisive importance in the economy of the collateral circulation. On the basis of these new elements, together of course with radiological and fibro-endoscopic data, it is possible to divide cases of portal hypertension into three groups depending on the type of collateral circulation operative on each occasion. These types present differing anatomcfunctional features and clinical physiognomy, a factor of great practical importance especially for the purpose of establishing indications for the portacaval anastomosis of choice, and possibly prophylactic intervention.


Acute changes of the Na-K-ATPase-activity in plasmamembranes of the isolated, cell-free perfused rat kidney.

The Na-K-ATPase-activity of the isolated, cell-free perfused rat kidney was investigated. Na-load and perfusion rate could be varied independently. The ATPase-assay was carried out on plasmamembranes (700-1200 g fraction) isolated and purified after 60 min perfusion. Both the specific Na-K-ATPase-activity and the fractional Na-reabsorption showed an indirect correlation with the Na-load as well as with the perfusion rate. The Na-K-ATPase-activity of plasmamembranes obtained from the isolated kidney at physiological Na-load fits well to control values prepared from unperfused control kidneys. The present data are in accord with the results of earlier micropuncture experiments, where acute saline loading diminished the rate of Na-absorption.


Persisting changes in sensory and motor activity of a muscle following its reflex activation.

A study was made to see if contractions induced by reflex action were followed by an elevation in muscle afferent activity comparable to the post-contraction sensory discharge (PCSD) that occurs after stimulation of the ventral roots. In lightly anesthetized cats, extension reflexes of the triceps surae muscle in the otherwise denervated leg were induced by contralateral stimulation of the popliteal nerve or footpad. The reflex response was followed by an elevated afferent discharge, which disappeared after a brief stretch, as is characteristic also of the PCSD. Crossed-extension responses in fusimotor activity unaccompanied by contraction of the gross muscle were also succeeded by an elevation in sensory discharge and an increased sensitivity to a vibratory stimulus applied to the tendon. Pinna reflexes had a similar effect. In the presence of the elevated discharge, alpha-motoneurons demonstrated increased sensitivity to vibration stimuli. It is suggested that the increase in spindle sensitivity may help stabilize the muscle against minor pertubations in length, at least when background fusimotor activity is relatively quiet.


[The development of the larvae of the sheep nasopharyngeal botfly, oestrus ovis l. (diptera, oestridae)].

In order to find out factors affecting the presence of two generations of O. ovis in the southern parts of its distribution area there were conducted regular measurements of the body length of the Ist instar (3718 specimens) and weight of the IInd and IIId instar larvae (1460 specimens). These observations were carried out in two herds of sheep in low-lying and foothill zones of Aserbaijan. Statistical analysis has shown the following: a) the growth inhibition, which is especially distinct in autumn-spring generation, takes place in the Ist instar larvae 1.76-2.20 mm long inhabiting the walls of the nasal cavity and concha (their average body length at hatching is 1.08 plus or minus 0.004 mm); the inhibition is associated with interpopulation relations and apparently does not depend on the date of its beginning and can last from 6 to 7 months; c) after the growth resumption the development continues uninterruptedly up to the moulting; the inhibition is also possible at the beginning of the 2nd instar and then the development proceeds without any intervals up to the complete maturation of larvae. The total number of the Ist instar larvae in the bot fly populations is two times higher than that of the IInd and III-instars. This points to a high mortality of larvae during the Ist instar developmental period.


The electrocardiogram in acute pulmonary embolism.

Electrocardiograms of 90 patients with arteriographically documented acute submassive or massive pulmonary embolism and no associated cardiac or pulmonary disease were studied. Patients were derived from the Urokinase-Pulmonary Embolism Trial National Cooperative Study. In massive embolism, the electrocardiogram was normal in 6 per cent (3 of 50) of patients. With submassive embolism, 23 per cent of patients (9 of 40) had a normal electrocardiogram. Since one or more of the traditional manifestations of acute cor pulmonale (S1Q3T3, right bundle branch block, P pulmonale, or right axis deviation) occurred in only 26 per cent of patients, one could not rely exclusively upon these electrocardiographic abnormalities for the diagnosis of pulmonary embolism. The most common electrocardiographic abnormalities were nonspecific T wave changes which occurred in 42 per cent of patients and nonspecific abnormalities (elevation or depression) of the RST segment which occurred in 41 per cent of patients. Left axis deviation occurring in 7 per cent of the patients was as frequent as right axis deviation. Low voltage QRS complexes, previously undescribed in pulmonary embolism, occurred in 6 per cent of patients. None of the patients had atrial flutter or atrial fibrillation, which appears to occur more typically in patients with pulmonary embolism who have preexistent cardiac disease. All of the varieties of electrocardiographic abnormalities disappeared in some of the patients by 2 wk. Inversion of the T wave was the most persistent abnormality. Larger defects on the lung scan or pulmonary arteriogram occurred in patients with various abnormalities on the electrocardiogram than in patients with normal electrocardiograms. The pulmonary arterial mean pressure and/or right ventricular end-diastolic pressure was significantly higher in patients with several varieties of abnormal electrocardiograms, although the partial pressure of oxygen in arterial blood, in general, did not differ from that in patients with normal electrocardiograms. These hemodynamic correlations, made for the first time in patients, suggest that acute ventricular dilatation, possibly in combination with hypoxemia, is a causative factor of the electrocardiographic changes in acute massive or submassive pulmonary embolism.


Typhoid cardiac involvement.

Three cases of typhoid cardiac complications are reported. Salmonella typhi was the aetiological agent in all three; The discovery of 3 patients over a period of 18 months merits special interest, especially since typhoid fever is endemic in the area concerned. The significance of the complication reported here is further enhanced by absence of similar specific cases in the English literature dealing with cardiac salmonellosis. One of the cases described in this article, the only fatality of the series, developed a rhythm disturbance identical with that of a patient whose myocardial abscess was due to Salmonella typhimurium. The rarity of typhoid cardiac complications may be deceptive; The septicaemic disturbance may mask it and one must note that cardiac salmonellosis is reported from developed countries, where typhoid fever is a rarity.


Pulmonary stenosis and impaired myocardial function.

Two young Black female patients with pulmonary valve stenosis and intact ventricular septa are presented in protracted congestive cardiac failure with severe tricuspid insufficiency and, in one, atrial fibrillation. Right ventricular systolic dysfunction was manifested by peak systolic pressures below systemic level, raised end-diastolic pressures and low cardiac output, but without right-to-left shunt. These findings are in strong contrast to those found in most patients with pulmonary stenosis of long standing, where persistent impairment of right ventricular function is diastolic with a high end-diastolic pressure and reversal of an interatrial shunt which result from poor right ventricular compliance. Evidence of left ventricular dysfunction was also present in both cases. Protracted heart failure in these patients is believed to have been the result of coincidental cardiomyopathy in a racial group highly predisposed to this disorder. A diagnostic appreciation of this phenomenon is important in the evaluation of heart disease in the Black, since cardiomyopathy may modify or even mask the features of the underlying disorder.


Clinical and roentgenographic evaluation of endoscopic suspension of the vesical neck for urinary incontinence.

Forty-four consecutive patients, without residual urine or infecttion, were operated upon for urinary stress incontinence by endoscopie suspension of the vesical neck. All patients had preoperative and post-operative lateral view cystograms which showed after operation substantial upward and forward elevation of the urethrovesical junction as measured by the posterior urethrovesical angle, the angle of urethral inclination, and elevation of the base of bladder. Despite a significant history of an urgency component to their incontinence in many patients, as well as 44 previous operations in 22 of the patients, a carefully performed cystometrogram and cholinergic sensitivity test were not helpful in detecting abnormalities of the detrusor muscle. Fourty-one patients were cured, and in three others, the condition was not cured, the failure group. The lateral view cystogram was the most diagnostic test in that two in the failure group did not show preoperatively the roentgenographic hallmark of stress  urinary incontinence.


A new synthetic absorbable suture.

The experimental synthetic absorbable suture XLG is a co-polymer derived from lactic and glycolic acid through lactide and glycolide intermediates. Determinations of the rate of loss of strength in various sites in experiments on dogs show that XLG loses strength slightly more rapidly than does chromic catgut. An exception is sutures suspended in the stomach, where catgut quickly loses strength, whereas XLG maintained strength similar to that at other implantation sites.


A preliminary report on the use of a chronic peritoneal fistula for endoscopic examination of ovine ovarian activity.

A description is given of a technique devised to allow repeated endoscopic observations to be made of ovarian changes occurring during an oestrus cycle in the ewe.  The use of a chronic peritoneal fistula is illustrated and the methods used to maintain its patency discussed.  The technique was carried out in a small number of ewes and the ovarian changes occurring in an ovary of one of these ewes, over a 22 day period, are shown in photographic and diagrammatic form.


Urinary gonadotrophins in the Sertoli-Cell-only syndrome.

In order to study the hypophyseal-testicular axis in males with complete absence of germinal epithelium, the urinary total hypophyseal gonadotrophins (HG), urinary follicle stimulating hormone (FSH) and urinary luteinizing hormone (LH) were measured by specific bioassays in 12 males with classical Sertoli-cell-only syndrome and compared with HG, FSH, and LH in normal and castrated men. HG and FSH were significantly higher than HG and FSH in normal men (P smaller than 0.0025, P smaller than 0.0005, respectively), but significantly lower than in castrated men (P less than 0.001, P less than 0.01, respectively). LH was not different from LH of normal men, but significantly lower than in castrated men (P less than 0.0005). All patients had normal excretion of androgen metabolites (androsterone plus aetiocholanolone) but a dexamethasone suppression test, performed in 8 subjects, revealed that in 2 cases of testicular origin, the values were below the normal range. The excretion of oestrogens was within the normal range. The presented data support the concept that the germinal epithelium produces a substance capable of inhibiting FSH secretion from the hypophyses, the Sertoli cell itself, however, having a basal production of this inhibitor. The finding of low excretion of testicular androgen metabolites in some of the patients and normal urinary LH, indicates that disturbances in the LH-testosterone feedback mechanism in such patients may occur and that the previous concept of isolated defects of spermatogenesis in all such patients was erroneous.


Laparoscopy in endocrine and genetic disorders of the gonads.

The guideline of this work is to examine the diagnostic value of laparoscopy in some endocrine and genetic syndromes connected with gonadal pathology or abnormal sexual development in relation to the various hormonal and cytogenetic techniques usually utilized by the endocrinologist. After a brief introduction, Chapter 1 stresses our purpose to examine the advantages and limits of laparoscopy by investigating the most significant cases that came to our observation. Attention is drawn in Chapter 2 to the application of laparoscopy in a particular form of amenorrhea (uterus absence), admittedly of great endocrinologic interest. In particular, the results obtained by the application of this technique are reported in cases of uterus agenesis and the testicular feminization syndrome. The clinical, cytogenetic and hormonologic study of one case of uterus agenesis is reported together with that of four cases of the testicular feminization syndrome with the view to discussing the differential diagnosis between these two syndromes and the particular diagnostic problems connected with the latter. Our conclusion is that laparoscopy, although usually having only a confirmatory value in these syndromes, may assume in certain cases major diagnostic importance (the post-puberal testicular feminization syndrome without palpable gonads, the pre-puberal testicular feminization syndrome, rare cases of the testicular feminization syndrome associated with chromosome aberrations). Chapter 3 discusses the application of laparoscopy in female gonadal dysgenesis on the basis of three cases of Turner's syndrome with different diagnostic problems and one of pure gonadal dysgenesis. It is stressed that, although in the majority of cases of Turner's syndrome the laparoscopic findings are quite characteristic (streak gonads), in some cases the gonads may also present the aspect of hypoplastic ovaries. This finding is observed in two cases of Turner's syndrome with XO/XX mosaicism, of which one case with spontaneous menarche. The diagnostic value of laparoscopy is discussed in the cases of pure gonadal dysgenesis and in variants of Turner's syndrome chromatin-positive with X chromosome partial monosomy and chromatin-negative with a Y chromosome. Our conclusion is that laparoscopy assumes particular diagnostic value in cases of pure gonadal dysgenesis without chromosome aberrations, because chromosome aberrations may be absent in this syndrome, as well as in chromatin-negative variants with a Y chromosome because the gonads and gonaducts differentiation may be variable.


High voltage QRS complexes in children with innocent, "vibratory" heart murmur.

The electrocardiographic findings in 86 four-year-old children with innocent murmur of the so-called vibratory type are described. The electrical axis, the Q, R and S waves in the right and left precordial leads were compared with figures from the literature and with a control group of 4-year-old children without signs of somatic disease. The most significant findings were higher amplitudes of Q and R waves over the left precordium in children with vibratory murmur. Many children in this group had amplitudes of these waves exceeding accepted normal limits. There were no findings compatible with organic heart disease and the difference as compared to normal values and controls is probably a matter of variations in the position of the heart.


Bacterial endocarditis in idiopathic hypertrophic subaortic stenosis.

Bacterial endocarditis complicating idiopathic hypertrophic subaortic stenosis (IHSS) is uncommon but endocarditis may be the first clinical manifestation of IHSS. In this report of such a case, the aortic and the mitral valves were the sites of the bacterial infection. Many chordae tendineae to the mitral valve were ruptured from the extension of the infectious process. The endothelial lesions, which served as the seat for the bacterial infection on the anterior mitral leaflet, likely resulted from its abutting action against the septal prominence. Damage to the aortic valve leaflet may have resulted from abnormal valve motion caused by IHSS and created an environment conducive to endocarditis. This patient developed aortic insufficiency during the course of bacterial endocarditis, suggesting that the occasional association of aortic insufficiency in patients with IHSS may be secondary to healed endocarditis of the aortic valve.


Cardiac pathology after aortic valve replacement using Hufnagel trileaflet prostheses: a study of 20 necropsy patients.

Necropsy observations are described in 20 patients dying between October 1967 and March 1973 after replacement of the aortic valve with a Hufnagel trileaflet prosthesis. Seven patients died within two months of operation and 13, between 2.1 and 58 months (average 22). Four of seven patients dying early had extensive prosthetic thrombus causing obstruction of one coronary arterial ostium in each. Of the 12 patients surviving six months or longer after valve replacement, death in eight resulted from degeneration (tearing) of the prosthetic cusps causing severe aortic regurgitation in each, and from thrombosis of the prostheses in three, probably causing prosthetic stenosis and defintely causing narrowing of at least one coronary ostium. Thus, prosthetic degeneration or thrombosis caused death in 11 of the 12 patients surviving six months or longer. In conclusion, this trileaflet aortic prosthesis, although similar in design to the normal aortic valve, is composed of materials not durable enough to withstand the stresses created by blood flow in this position.


Uchida tubal sterilization.

The methods which seek to close the tubes by tissue necrosis may be responsible for subsequent pathologic disorders. These sequelae often explain postoperative repeat uterine pregnancy, ectopic gestation, and other serious complications. In addition to Uchida sterilization, the Uchida fimbriectomy is recommended for use in the immediate postpartum period and at cesarean section because of its many advantages.


The acute effect of hysterectomy on ovarian function.

The acute effect of abdominal and vaginal hysterectomy on the ovarian production of estradiol-17-beta (E2) and progesterone (P) was studied in a group of patients undergoing hysterectomy for various gynecologic indications. Plasma levels of steroids as well as gonadotropins (FSH and LH) were measured by RIA. There was a significant but transient drop of plasma E2 during the follicular phase and of plasma E2 and P during the luteal phase, following hysterectomy. No significant variations in the steroid plasma levels were found in a control group of patients undergoing laparoscopy for diagnostic purposes. No changes were noted in either group in the plasma levels of gonadotropins. The changes seen appear to be related to the surgical manipulation in the ovarian region rather than to the type of hysterectomy performed or the stress of surgery.


Effect of exogenous dehydroepiandrosterone upon the fetoplacental biosynthesis of estrogens and its effect upon uterine blood flow in the term pregnant ewe.

The fetoplacental unit of the ewe is capable of increasing the biosynthesis of estrogens following the exogenous administration of DHEA to the fetus. The maximum concentrations of estrogens appeared approximately 30 minutes after the administration of DHEA. Uterine blood flow in the pregnant ewe increased approximately 90 minutes after the maximum concentrations of estrogens were noted. The administration of DHEA to the nonpregnant, ovariectomized ewe did not elicit estrogen biosynthesis or changes in uterine blood flow. Isotope experiments in the pregnant ewe demonstrated the incorporation of DHEA into urinary estradiol.


Electromicrosurgery and salpingostomy: long-term results.

It seems that a technique which is as atraumatic and as bloodless as possible gives good results. This "atraumatic and bloodless" surgery is possible by means of electrosurgery, refined to a form of microsurgery. The author presents his long-term results (observations over 8 years). Pregnancies: 63 per cent; resulting in 24 per cent ectopic pregnancies and 36 per cent live births; 3 per cent pregnant pregnant at present.


Effect of acute pelvic inflammatory disease on fertility.

A total of 415 women treated for laparoscopically verified pelvic inflammatory disease (PID) were reviewed after 9.5 years. Of these, 88 (21.2 per cent) were involuntarily childless after one or more infection; in 72 cases (17.3 per cent) this was due to tubal obstruction; 263 (63.4 per cent) women became pregnant; 64 (15.4 per cent) were voluntarily childless. Tubal occlusion was diagnosed after one infection in 12.8 per cent, after two infections in 35.5 per cent, and after three or more infections in 75 per cent of the women. Tubal occlusion was more common after nongonorrheal than after gonorrheal salpingitis. Infertility varied with the inflammatory changes seen at laparoscopy. The ratio between ectopic and intrauterine pregnancies after the infections was 1/24. Chronic abdominal pain was reported by 18.1 per cent of the women. Corresponding findings in 100 healthy control subjects were: involuntary childlessness in three despite normal Fallopian tubes, one ectopic in 147 intrauterine pregnancies, and chronic abdominal pain in five cases.


Rural health and the role of occupational therapy.

Through the use of activities which are common to the rural culture, the occupational therapist may make a contribution in rural health services. Although the rural resident suffers psychiatric and physical disorders similar to his urban neighbor, the availability and accessibility of medical services are more limited. The isolation of rural life which makes it unattractive to health and other professionals also accentuates the problems of the rural patient. The rural health clinic has been one solution to the lack of medical services. It is proposed that the occupational therapist select farm activities which are common to the patient, and adapt and direct those activities so that the patient can again function in his rural environment.


Effects of ouabain and amiloride on Na pathways in turtle bladders.

In a Na-rich bathing system, addition of amiloride to the mucosal fluid of turtle bladders produces decreased in the transepithelial potential difference (PD), short-circuiting current (I-sc), and conductance. Removal of amiloride results in complete reversal of these changes; and this reversibility is incomplete in amiloride-blocked bladders exposed to ouabain. In a Na-free bathing system, step increased in mucosal [Na] evoke rapid initial spikes in PD, Isc, and conductance, the magnitudes of which are independent of prior ouabain treatment. After these spikes, PD and Isc in the ouabain-treated bladder rapidly decay, while conductance remains unchanged and high. This unchanging conductance, plus the fact that ouabain inhibits half the microsomal (Na plus K). ATPase of this tissue within 1 min, suggests that ouabain inhibits Na pumping without changing tissue conductance. The delayed decrease in conductance (beginning 30 min after ouabain addition), a nonspecific and secondary effect of ouabain, is due to a concomitant collapse of the intercellular channels.


Continuous positive airway pressure breathing (CPAP).  Apparatus for use in neonates or adults.

CPAP is a technique of respiratory care which was originally described in the management of the respiratory distress syndrome of the newborn and later in the post operative management of the cardiac infant following surgery. It has potential value in the respiratory management of older children and adults. Apparatus is described suitable for the application of continuous airway pressure during spontaneous ventilation via endotracheal tube in either neonates or adults. The inspired oxygen content is adjustable and the fresh gas warmed and humidified. High and low pressure alarms are not considered necessary. A head-harness is described for the application of CPAP in neonates via twin nasal or nasopharyngeal tubes. This equipment may be obtained from Messrs. Lusterlite Products Limited, 56 Devon Road, Leeds 2.


Lipid droplet accumulation in cardiac muscle cells of the bat: potential auto-toxicity of the cardiac sympathetic innervation.

The previously described ability of reserpine and parachlorophenylalanine to induce the accumulation of lipid droplets in ventricular cardiac muscle cells of the bat was investigated. Lipid droplet accumulation was assessed qualitatively by light microscopy and quantitatively by morphometric analysis of electron micrographs. An hypothesis that the action of the drugs was an indirect one, mediated by the cardiac adrenergic innervation, was framed and tested. Lipid droplet accumulation occurred during a time of intense sympathetic activity, that of arousal from hibernation. The ability of the two drugs to produce the effect was antagonized by prior sympathetectomy with 6-hydroxy-dopamine. The effect was mimicked by administration of exogenous norepinephrine together with inhibitors of its catabolic enzymes, monoamine oxidase and catechol-o-methyl transferase. These observations are all consistent with the initial hypothesis and raise the possibility that endogenous norepinephrine in the cardiac sympathetic innervation might be, at least potentially, auto-toxic.


In-vitro investigation of a new neuromuscular relaxant, AH8165.

AH8165 was compared with other neuromuscular relaxants in an in-vitro rat phrenic nerve-diaphragm preparation. Concentrations of 6-10 mug/ml AH8165 produced progressive decreases in strength of concentration. AH8165 was 0.1 times as potent as d-tubocurarine, and its effects were more rapidly reversed by washing. The times to recovery from 90 per cent blockade were the same for succinylcholine and AH8165, but the time to recovery from 50 per cent blockade was shorter for succinylcholine than for AH8165. Neostigmine reversed blockade induced by d-tubocurarine to 80 per cent of control, while it reversed comparable blockade induced by AH8165 to only 40 per cent of control. Doses of 0.5 to 2 mug AH8165 produced contracture and increased the force of contraction of the superfused chick biventer cervicis muscle preparation. Doses of 8 to 32 mug produced decreased contracture followed by diminution of the strength of contraction. The authors conclude that AH8165 in low concentrations has a depolarizing action, which is obscured by nondepolarizing effects in higher concentrations. (Key words: Neuromuscular relaxants: AH8165.)


Cardiovascular effects of intraperitoneal insufflation with carbon dioxide and nitrous oxide in the dog.

Cardiovascular changes caused by intraperitoneal insufflation with CO2 or N2O were measured in 15 mongrel dogs. Moderate progressive increases in intra-abdominal pressure (to 40 mm Hg) with either gas produced increases in mean arterial, right atrial, pleural, and femoral-vein pressures. Cardiac output and inferior vena caval flow were momentarily increased following the commencement of insufflation. However, both flows decreased precipitously as insufflation pressure was increased. At an intra-abdominal pressure of 40 mm Hg cardiac output and inferior vena caval flow were reduced more than 60 per cent in most cases. Peripheral resistance increased by approximately 200 per cent. Upon sudden release of abdominal pressure cardiac output and inferior vana caval flow increased but then returned to pre-insufflation values within seconds. Directly measured right atrial pressure increased with increasing insufflation pressure, but calculated transmural right atrial pressure decreased with the increase in intra-abdominal pressure. Insufflation with CO2 produced significant increases in PaCO2. However, cardiostimulatory effects due to elevated blood CO2 levels were not seen. The data from this study indicate that intraperitoneal insufflation produces serious hemodynamic alterations which are manifested by low cardiac output and elevated total peripheral resistance. In addition, directly measured right atrial pressure cannot be used clinically as an indicator of venous return to the heart since it reflects a composite of pleural and intra-abdominal insufflation pressure. (Key words: Anesthetics, gases, nitrous oxide; Carbon dioxide, intraperitoneal; Surgery, intraperitoneal insufflation; Heart, function, intraperitoneal insufflation.).


Acute hypotension during laparoscopy: a case report.

A 27-year-old, generally healthy, woman became acutely hypotensive during a diagnostic laparoscopy. Her blood pressure returned to preoperative levels and remained stable after decrease in intra-abdominal pressure. The author suggests five possible causes of hypotension during laparoscopy.


Influence of antibody-mediated immune suppression on clinical, viral, and immune responses to swine influenza infection.

Antibody-mediated immune suppression occurred when newborn pigs with naturally acquired passive antibody were exposed to seine influenza virus. Frequency and relative ease of recovery of virus from nasal secretions were inversely related to the concentration of specific passive antibody existing at time of exposure. Severe overt respiratory signs during the acute stages of the disease were observed only in pigs with low passive antibody concentrations. The concentration of passive antibody at the time of exposure determined the immune status of the pig during the convalescent stage of disease. Infection could occur in the presence of high passive antibody concentrations, but the pig was not immunologically stimulated. Reexposure after the decay of passive antibody produced primary immune respone, severe clinical reinfection, and recovery of virus from nasal secretions for a period of time similar to that seen in pigs having their first exposure. Infection of newborn pigs with low passive antibody concentrations led to immunologic priming. A second exposure to virus produced a secondary immune response, mild clinical disease, and shortened time during which virus was recovered from nasal secretions. The relevance of these studies for the practice of vaccination or infections of the dam before parturition so that the neonate will have specific passive immunity is discussed.


Distribution and seasonal prevalence of bovine lungworms in selected areas in western Montana.

In a survey (Oct, 1967, through Aug, 1969) of the distribution and seasonal prevalence of bovine lungworms in beef herds in 3 western Montana counties (Park, Gallatin, and Ravalli), infected cattle were detected in 27 of 35 herds. Results of 2,125 fecal examinations indicated that 7.8% of all cattle were infected with Dictyocaulus viviparus. Lungworms were found in 6.6% of 1,038 calves (less than 12 months old), 11.5% of 755 yearlings (1 to 2 years old), and 3.3% of 332 cows (more than 2 years old). A distinct seasonal pattern existed in calf infections, with initial passage of larvae beginning in late July or August and persisting until November or December. Exposure of calves in 1 portion of the study area apparently was delayed until November, with patent infections continuing through March. Spontaneous loss of infections normally occurred after 3 to 4 months of patency. The mean rate of larval excretion in the feces of all infected cattle was 0.37 larvae per gram of feces (l/g). Larval output was relatively uniform within each geographic area regardless of host age, but varied considerably from one portion of the study area to another. The potential role of wild ruminants (deer, elk, and moose) in the epizootiology of D viviparus is discussed with respect to western range utilization practices.


Gonorrhea: recommended treatment schedules.

Appropriate drugs and their proper use are described for the care of adult and pediatric patients with uncomplicated and complicated gonococcal infections in men and women. Special diagnostic considerations in some cases are indicated. These recommendations were established by the Venereal Disease Control Advisory Committee after deliberation with therapy consultants.


A hypothesis to account for the Mary Walker phenomenon.

Certain myasthenic patients show that their skeletal muscles produce a substance during ischemic exercise which, when released into the circulation, increases muscle weakness. This phenomenon has been named after Mary Walker, the physician who popularized its demonstration. It is suggested that ischemic exercise produces lactic acid that binds calcium, reducing ionized and total serum calcium. The decreased calcium has an adverse effect on skeletal muscle function especially in patients with myasthenia gravis, where neuromuscular function is often so precarious it may be interrupted by weak inhibitors.


Primary abdominal wall abscess diagnosed by ultrasound.

Ultrasound was used to diagnose and demonstrate an anterior abdominal wall abscess of unknown cause in a diabetic woman. This condition may mimic major abdominal disorders and, in our patient, appeared as acute cholecystitis. The use of ultrasound is simple, quick, painless, and without known adverse side effects. The diagnosis may be more easily and accurately established than by routine roentgenography.


Diphenylhydantoin. Blocking of levodopa effects.

Diphenylhydantoin (DPH) diminished the therapeutic effects of levodopa both in patients with parkinsonism and in patients with chronic manganese poisoning, as well as the levodopa-dependent dyskinesia for which the former were selected. In patients with Huntington chorea, it enhanced chorea and mental agitation and, thus, failed to conform with the postulated pharmacological reciprocity between Parkinson disease and Huntington chorea. These findings are in agreement with experiments done in animals in which DPH blocked a neuronal response to dopamine.


Mitochondrial adenosine triphosphatase. Location of sulfhydryl groups and disulfide bonds in soluble enzyme from beef heart.

The soluble beef heart mitochondrial ATPase (F1) contains eight sulfhydryl groups and two disulfide bonds. N-Ethylmaleimide has been used to radioactively label the sulfhydryl groups before and after cleavage of the disulfide bonds by dithiothreitol. After subjecting the labeled protein to polyacrylamide gel electrophoresis in sodium dodecyl sulfate and measuring radioactivity in each of the separated subunits the location of all the sulfhydryl groups and the disulfide bonds may be specified. The conclusions are supported by direct examination of depolymerized, unreduced, enzyme by polyacrylamide gel electrophoresis. The results also indicate that current ideas regarding the overall subunit structure of this enzyme may be incorrect, and this is discussed in light of new data presented here.


Structural proteins of dogfish skeletal muscle.

As part of a study on the evolutionary aspects of control mechanisms, a number of structural muscle components from the Pacific dogfish (Squalus acanthias) are described. These include troponin, tropomyosin, actin, and myosin. Troponin (mol wt 108.000) was resolved into its constitutive subunits, repeated by a 20,500 mol wt fragment which binds 2 mol of Ca2+/mol with a KDiss of 0.91 mum, and an inhibitory component of 30,000 and a 58,000 component which are necessary for the calcium sensitivity of actomyosin ATPase. Tropomyosin and actin share many properties with their counterparts from higher vertebrates. Proteins similar to parvalbumins, i.e., the low molecular weight calcium-binding proteins widely distributed in fish, amphibians, and mammalian muscle, could be generated from troponin and its calcium-binding subunit by limited proteolysis. The appearance of immunological cross-reactivity and other similar features suggested some identity, but differences in the amino acid analysis exclude the possiblity that parvalbumins occur as breakdown products of troponin. The close relationship between parvalbumins and the calcium-binding subunit brings additional evidence that these proteins have arisen through divergent evolution.


Neural regulation of mammalian fast and slow muscle myosins: an electrophoretic analysis.

Mammalian nerves to fast and slow muscles have the remarkable property of changing the speed of contraction of muscles following cross-reinnervation. The biochemical basis of speed transformation is the change in myosin in ATPase activity. This paper provides electrophoretic evidence for structural changes in myosin from cross-reinnervated muscles. A method is described for the separation of intact fast and slow muscle myosins by polyacrylamide gel electrophoresis. This method utilizes the fact that ATP and its analogs prevent the formation of myosin polymers in low ionic strength buffers. In this system, normal fast muscle myosin has a higher electrophoretic mobility than slow muscle myosin. Normal rat soleus myosin has a major slow and a minor fast component due to two populations of muscle fibers. The same muscle cross-reinnervated by a fast muscle nerve shows only the fast component, The normal, homogeneous fast extensor digitorum longus muscle has only the electrophoretically fast myosin, but following cross-reinnervation it shows both fast and slow components. These results suggest that mammalian motor nerves can induce or suppress the expression of genes that code for fast and slow skeletal muscle myosins.


Functional relationship between the ADP/ATP-carrier and the F1-ATPase in mitochondria.

1. The distribution of labeled and unlabeled adenine-nucleotides inside and outside mitochondria was followed after addition of [14C]ADP to rat liver mitochondria. Two types of mitochondria were used: 1, respiring mitochondria which were carrying out oxidative phosphorylation and which had been replenished in ATP by incubation in a medium supplemented with succinate and phosphate; 2, non-respiring mitochondria which had been partially depleted of ATP by incubation in a medium supplemented with rotenone and phosphate. During the first minute following addition of [14C]ADP to the respiring mitochondria, the pre-existing intramitochondrial (internal) [12C]ATP was released into the medium and replaced by newly synthesized [14C]ATP. No [14C]ADP accumulated in the mitochondria. It is suggested that extramitochondrial (external) ADP entering respiring mitochondria in exchange for internal ATP is phosphorylated to ATP before its complete release in the matrix space. In non-respiring mitochondria, the entry of [14C]ADP into the mitochondria was accompanied by the appearance in the external space of [12C]ADP and [12C]ATP, with a marked predominance of [12C]ADP. Thus in non-respiring mitochondria, the residual internal ATP is dephosphorylated to ADP in the inner membrane before being released outside the mitochondria. 2. When mitochondria were incubated with glutamate, ADP and [32P]phosphate, the [32P]ATP which accumulated in the matrix space became rapidly labeled in both the P gamma and P beta groups of the ATP, due to the presence of a transphosphorylation system in the mitochondrial matrix. The [32P]ATP which accumulated outside the mitochondria was also labeled in the P beta group, although less rapidly than the internal ATP. Our data show that a large fraction (75-80%) of the ATP produced by phosphorylation of added ADP within the inner mitochondrial membrane is released into the matrix space before being transported out from the mitochondria; only a small part (20-25%) is released directly outside the mitochondria without penetrating the matrix space. 3. In respiring and phosphorylating mitochondria, the value of the Km of the ADP-carrier for external ADP was 2-4 times lower than its value in non-respiring and non-phosphorylating mitochondria. 4. The above experimental data are discussed with reference to the topological and functional relationships between the ADP-carrier and the oxidative phosphorylation complex in the inner mitochondrial membrane. They strongly suggest that the ADP-carrier comes to the close neighbourhood of the ATP synthetase on the matrix side of the inner membrane.


Comparison of proteoglycans from bovine articular cartilage.

Four bovine articular cartilages have been compared with regard to the chemical composition of the whole cartilages, the amount of proteoglycan selectively extracted with 3 M MGCl2 or with 3 M guanidine-HCl, and the compositions and physical properties of the isolated proteoglycans. The whole cartilages differ but slightly in composition. Occipital condylar cartilage, a thin cartilage from the smallest joint, contains 4% more collagen and proportionately less proteoglycan than proximal humeral, the thickest cartilage from the largest joint. Each cartilage contains a pool of proteoglycan that resists extraction with 3 M MgCl2 but is extracted with 3 M guanidine-HCl. The proteoglycan extracted from each cartilage with 3 M guanidine-HCl contains a high molecular weight proteoglycan-collagen complex demonstrated by analytical ultracentrifugation and by the turbidity of its visible and ultra-violet spectra. The four cartilages appear to differ most remarkably in the fraction of total proteoglycan extracted from each as proteoglycan-collagen complex.


A new foam for support of the physically handicapped.

This article is intended to inform professionals in the fields of rehabilitation medicine and biomedical engineering of some of the more significant properities of Temper Foam, a new foam for support of the physically handicapped. The unique properties of this foam are explained in terms of its microstructure, illustrated by scanning electron micrographs and by mechanical tests using an Instron Materials Testing machine. Fourteen non-ambulatory patients with Duchenne muscular dystrophy are using this foam in custom-fitted contoured support systems. Normally, these children show increasing signs of discomfort and skin breakdown due to their inability to alternate areas of weight bearing and high local pressure such as the ischial tuberosities. In our support systems lined with Temper Foam these problems have completely dissappeared. We find that Temper Foam types T-36, T-38 and -41 are ideally suited for patients weighing up to 35 kg. T-41 may be used for patients weighing up to 70 kg.


Glomerulonephritis associated with Coxiella burnetii endocarditis.

A patient with endocarditis associated with chronic Coxiella burnetii infection is described in whom glomerulonephritis developed with granular deposits containing immunoglobulins and complement in the glomeruli. The serum was notable for the variety of circulating antibodies detected, which included antibodies directed against native DNA.


Circulating soluble antigens and antibody in schistosomiasis.

Some patients with Schistosoma mansoni and S. haematobium infections were shown to have soluble schistosomal antigen in their plasma. Antibody to this antigen and to other antigens of adult worms were also present. These findings may be relevant to schistosomal-related immune-complex disease, especially the nephrotic syndrome.


Effect of pinealectomy, gonadectomy, pCPA and pineal extracts on the rat parvocellular neurosecretory hypothalamic system; a fluorescence histochemical investigation.

Using the fluorescence histochemical technique, yellow autofluorescent granules were observed in neurones of the arcuate and ventromedial hypothalamic nuclei of the rat (type I cells). In the same nuclei, neurones could be demonstrated showing a formaldehyde-induced yellow fluorescence (type II cells). Microelectrophoresis and special staining methods applied to the pineal gland revealed the autofluorescent compound to be a protein containing a relatively high content of tryptophan. It is probable that the formaldehyde-induced yellow fluorescence is due to the presence of serotonin. In view of investigating a possible functional relationship between the pineal gland and the parvocellular hypothalamic nuclei mentioned, hypothalamic type I and type II cells, as well as autofluorescent and serotonin-containing pinealocytes, if present, were quantified under the following experimental conditions: (1) p-chlorophenylalanine (pCPA) administration, (2) castration, (3) pinealectomy, and (4) pinealectomy followed by substitution using rat and sheep pineal extract. Administration of pCPA caused a decrease in the number of type II and an increase in the number of type I cells, both in the pineal gland and the hypothalamic nuclei. Castration, in contrast, was followed by an increase in the number of autofluorescent pinealocytes, but a decrease of autofluorescent neurones in the hypothalamic nuclei (type I cells) while the number of serotonin-containing pinealocytes increased; decreasing in both hypothalamic nuclei. After pinealectomy the hypothalamic nuclei showed an increase of type I neurones, but a decrease of type II nerve cells. Pinealectomy followed by substitution using pineal extracts restored the number of type I and type II neurones to that normally found in the arcuate and ventromedial hypothalamic nuclei of control animals. The present investigation brings histological evidence of an influence exerted by the rat pineal gland on nuclei forming part of the hypothalamic hypophyseotropic area. The data obtained and some of the literature strongly suggest that the type II neurones, which probably contain serotonin, inhibit, in the same hypothalamic nuclei, the production of luteinizing hormone-releasing factor (LH-RF). As yet, the function of the autofluorescent compound present in the type I neurones is not known.


Glycolytic enzymes in resting spores and vegetative mycelia of Entomophthora pyriformis.

Specific activities of 14 enzymes of the Embden-Meyerhof-Parnas and pentose phosphate pathways were determined in extracts of resting spores and vegetative mycelia of Entomophthora pyriformis. All these enzymes were detected in mycelial extracts, whereas only nine were detectable in resting spore extracts. Activities of detectable spore enzymes were much lower than those of the corresponding mycelial enzymes with the exception of triosephosphate dehydrogenase, which had higher activity in spore extracts. The enzyme deficiencies noted point to the inability of either pathway to function in dormant spores.