Sterochemical preferences for curarimimetic neuromuscular junction blockade III: enantiomeric bisquaternary amines related to benzoquinonium as probes. Enantiomeric neuromuscular junction blocking agents, which are of the benzoquinonium type but which have a methyl group introduced adjacent to the quaternary moieties to provide an asymmetric center were synthesized and tested to determine whether the neuromuscular junction exhibits the relatively modest (R) greater than (S) superiority shown toward previously tested bisquaternaries. Testing included a mouse inclined screen assay and an in vivo cat hypoglossal nerve-tongue preparation, as well as standard estimations of anticholinesterase activity since the candidate compounds are known to have such a component in their activity spectrum. The observed 2:1 difference in blocking activity favoring the compound with an (R)-configuration is the same as that for previously tested bisquaternaries, both in direction and magnitude. Furthermore, it cannot be accounted for by preferential inhibition of acetylcholinesterase by the (S)-enantiomer. Absolute configurations of the enantiomers were assigned on the basis of comparison with compounds of known configuration.


Determination of iodochlorhydroxyquin and corticosteroids in pharmaceutical formulations. Iodochlorhydroxyquin was separated from various corticosteroids using an acetonitrile-diatomaceous earth column. Iodochlorhydroxyquin was eluted with cyclohexane, and the corticosteroid was eluted with chloroform. Iodochlorhydroxyquin was determined by both a UV absorbance method and a new compleximetric method using the nickel chelate of iodochlorhydroxyquin. The corticosteroid was determined by the blue tetrazolium and isoniazid procedures. The average percent recovery for these four methods was 100.8, 99.4,100.7, and 99.9, respectively, for 10 known mixtures. The standard deviation for the absorbance for 10 determinations of the nickel complex was 0.002 absorbance unit (0.31%). Various characteristics of the nickel and other complexes were evaluated, including the sensitivity, solubility, and wavelength of maximum absorbance in 14 different solvents. The analyses of 23 typical products are reported, for which the standard deviation, expressed as a percentage of the amount declared, was 1.31% for the UV, 1.34% for the compleximetric, 1.49% for the blue tetrazolium, and 1.22% for the isoniazid procedures. Methods of determination in the presence of interferences are discussed.


Effect of deoxyuridine coadministration on toxicity and antitumor activity of fluorouracil and floxuridine. The addition of deoxyuridine (UDR) to fluorouracil (FU) or floxuridine (5-fluoro-2' deoxyuridine) (FUDR) produced a substantial increase in their toxicity in BDF1 mice. Antitumor assays using sarcoma 180 tumor-bearing mice showed a concomitant increase in tumor growth inhibition for the nucleoside-drug combination over identical doses of the single drug. However, no significant increase in antitumor activity with the combination treatment was demonstrated when equitoxic doses were given. Additional support for the therapeutic equality of the single and combination drug regimens was the similarity of the therapeutic indexes for each treatment regimen involving either fluorouracil or floxuridine. The results suggested that any therapeutic benefit achieved with the combination therapy could be duplicated with either fluorouracil or floxuridine at a higher dose.


Effect of gentamicin on renal function in the rat. The functional correlates of gentamicin nephrotoxicity were studied in rats injected with gentamicin, 100 mg/kg b.wt. per day, for 1 to 6 days. After 4 days of injections, a significant decrease in urine osmolality was detected in association with a rise in blood urea nitrogen and serum creatinine. These changes became more pronounced after 5 days of injections. Injecting gentamicin for 3 to 5 days had no discernible effect on the capacity of the distal nephron to reabsorb solute free water during a saline load compared to the control group. Tubular secretion of para-aminohippurate (PAH) was significantly greater in rats injected with gentamicin for 4 to 5 days than in the control group. Increased uptake of PAH by renal cortical slices was detected after 1 day of gentamicin treatment and reached a peak effect after 5 days of injections. Probenecid completely inhibited the augmented PAH uptake. No stimulation of the organic base transport system was observed. Three days of gentamicin treatment did not alter intracellular water or electrolyte content of kidney cortical slices. The data suggest that gentamicin specifically stimulates the organic acid transport system and that this effect may represent an early functional correlate of gentamicin nephrotoxicity.


Influence of L-tryptophan on morphine analgesia, tolerance and physical dependence. Four hours after the acute administration of L-tryptophan (75 mg/kg) to either, nontolerant or morphine-tolerant mice, the antinociceptive effect of morphine was partially and significantly antagonized. Daily tryptophan administration to rats and mice during a 3-day morphine pellet implantation period increased the rates of both morphine tolerance development and development of physical dependence. The accelerating effect of tryptophan on tolerance and dependence development in mice was antagonized by pretreatment with the tryptophan hydroxylase inhibitor, p-chlorophenylalanine. Acute tryptophan administration (75 mg/kg) significantly increased mouse brain 5-hydroxytryptamine levels for at least 4 hours. Although chronic tryptophan treatment increased 5-hydroxytryptamine turnover in morphine-treated mice, no effect of chronic morphine or tryptophan treatment on the particulate tryptophan  hydroxylase activity of whole mouse brain was observed. Slight increases in tryptophan hydroxylase activity were observed in the caudate-putamen and septal areas of rat brain 3 and 6 days, respectively, after s.c. morphine pellet implantation. These and previous studies from our laboratory indicate that the development of morphine tolerance and dependence can be modified by agents affecting serotonergic mechanisms.


Effect of growth in lithium on ouabain binding, Na-K-ATPase and Na and K transport in hela cells. 1. HeLa cells were grown for 24 hr in growth medium in which part of the Na was replaced with Li. Ion contents, cell volumes and numbers, Na-K-ATPase and specific ouabain binding were measured. In some experiments the Na efflux and net Na transport was also measured. 2. Growth in Li caused a rise in the specific ouabain binding and membrane Na-K-ATPase of these cells. The Li concentrations in the cells necessary to produce this effect ranged up to 50 mM. 3. It is suggested that Li, like Na, acts on the genetic material of the cells to cause the production of more Na pumps within the membrane.


The interaction of ATP-analogues possessing a blocked gamma-phosphate group with the sodium pump in human red cells. 1. The (Na++K+)-ATPase of red cell membranes is unable to hydrolyse ATP-analogues in which the oxygen atom linking the beta- and gamma-phosphate groups is replaced by a minusCH2minus or minusNH-bridge. 2. In resealed ghosts both these ATP-analogues support K:K exchange but not Na:K exchange. ATP supports both modes of operation of the sodium pump, whereas neither occurs without any nucleotide. 3. These results support the hypothesis that ATP is needed as a cofactor for K:K exchange to occur, and make it extremely unlikely that phosphorylation from ATP is involved.


An investigation into the site of termination of static gamma fibres within muscle spindles of the cat peroneus longus muscle. 1. The distribution of static fusimotor fibres to intrafusal muscle fibres of cat peroneus longus muscle spindles was investigated using the glycogen-depletion technique of Edström & Kugelberg (1968). Single static gamma fibres were stimulation intermittently at high rates for 3 hr with the blood supply occluded for some of this time. Subsequently the portion of muscle containing the activated spindles was fixed, sectioned and stained for glycogen with the periodic acid-Schiff (PAS) method. 2. Ten static axons caused depletion in eleven spindles. In five of these the only glycogen-depleted fibres were nuclear chain fibres. In the other six spindles one nuclear bag fibre was depleted in addition to chain fibres and this was always the larger of the two within the spindle. 3. These results on a medium-sized hind limb muscle are compared with findings concerning the distribution of static gamma fibre axons previously investigated only in very small muscles. The results agree in showing that nearly all static gamma fibres innervate nuclear chain fibres but that in 50-75% of the times in which static gamma fibres innervate spindles the distribution is to bag fibres as well as to chain fibres. The interpretation to be put upon this is uncertain. One possibility with which the results from peroneus longus are consistent is that the bag fibres which are usually innervated by static axons are the 'intermediate' bag fibres whose ultrastructure has recently been shown to resemble that of chain fibres.


Control of thymidine kinase synthesis in IHD vaccinia virus-infected thymidine kinase-deficient LM cells. The synthesis of vaccinia virus-induced thymidine kinase is normally arrested several hours after infection. In thymidine kinase-deficient LM cells infected with IHD strain of vaccinia virus, arrest occurs whether or not viral DNA synthesis is inhibited. With virus inactivated by UV irradiation, enzyme synthesis takes place, but arrest is abolished. It is suggested that an early viral genetic function is responsible for the cessation of thymidine kinase synthesis.


Coexistent ipsilateral subclavian steal and thoracic outlet compression syndromes. In this report we shall describe a 34-year old woman who presented with signs and symptoms of ipsilateral subclavian steal and thoracic outlet compression syndromes. The diagnosis was confirmed by arteriography and measurement of nerve conduction velocities. Via a single approach, the first rib was resected, the left subclavian artery was ligated at its takeoff, and flow was re-established with a woven Dacron graft. Two years postoperatively, the woman has equal and full pulses and blood pressure bilaterally, good strength, and no evidence of muscle atrophy.


Common atrium associated with anomalous high insertion of the inferior vena cava. The case is reported of a 38-year-old housewife with a common atrium associated with anomalous high insertion of the inferior vena cava, absence of the coronary sinus, and drainage of a left hepatic vein into the left side of common atrium. Surgical repair was performed successfully. The procedure was closure of a nearly total atrial septal defect with a Dacron patch. Embroyologically, the development failure of the sinu-atrial fold appears to be the basic causal factor.


[Healing of the donor area after transplantation of splitted grafts]. The healing of wounds in the donor area of splitted grafts was observed. The transplants served to graft an already existing wound which had not healed spontaneously. 120 wounds sustained by 95 patients were examined either histologically or by Rebuck's method of skin fenestration. 30 wounds were dressed with ointment and tulle, in 55 cases the wound was covered with perforated cellophane and an Allen-Koch compression bandage. These patients also received 1500 mg Complain in 500 ml physiological saline solution daily, by continuous drip infusion, for three days postoperatively. In 35 patients the wounds were only covered with perforated cellophane, but without additional medication. The application of Complamin shortened the healing process to seven to ten days in contrast to the control group where the wounds took 15 to 17 days to heal.


[Plastic surgery in the treatment of obesity (author's transl)]. The treatment of obesity must be primarily conservative. In suitable cases, a surgical operation may logically supplement it. Indications for operation: Localised accumulation of fat, generalised adiposity resistant to therapy, overstretched, flabby areas of skin and soft parts after radical conservative weight reduction or pregnancy, and esthetic considerations. The plastic surgical possibilities of "dermolipectomy" are pointed out, with typical examples.


[Gastro-intestinal endoscopy - today and tomorrow (author's transl)]. Because of their optical and mechanical properties, glass fibers have fulfilled the requirements for a panendoscopy of the gastrointestinal tract. The upper digestive tract (as far as the lower knee of the duodenum) and the large intestine, including the terminal ileum can be examined routinely nowadays with minimal risk. Surgical diagnostic and therapeutic endoscopy open up new possibilities, i. e. polypectomy, endoscopic biopsy, extraction of foreign bodies, papillotomy etc. The development of gastrointestinal endoscopy demands reorientation in training and teaching, in practice and in hospital.


[Thrombolytic treatment with urokinase (author's transl)]. Compared with streptokinase, thrombolytic treatment with urokinase has the advantages of being better tolerated and of practically unlimited applicability. Its disadvantage is the high cost. A good lytic action can be obtained with a dosage of 150,000 Ploug Units/12 hours for a duration of lysis of 8-14 days combined with heparin, the therapy being monitored by determination of the products of fibrinolysis. This dosage is not possible if the time factor plays a decisive role in the success of the treatment, e.g. in myocardial infarction. Urokinase is indicated when streptokinase cannot be used, or if continuation of the streptokinase therapy is necessary because of extensive thromboses.


Gamma-ray inactivation of conidia from heterokaryons of Neurospora crassa containing UV-sensitive mutations. Two-component heterokaryons were formed with the fungus Neurospora crassa. The UV-sensitive mutations upr-I, uvs-4, and uvs-6 were utilized. Conidia produced by these heterokaryons were exposed to gamma-rays and survival curves were established for the three conidial fractions produced by each heterokaryon. Results showed that upr-I, when included in only one nuclear component, did not affect the sensitivity of any conidial fraction; however, when included in both components, all three conidial fractions exhibited two- to four-fold decreases in survival at the 30 krad exposure. The uvs-4 mutation, when included in one or both components, did not increase the sensitivity of any conidial fraction and appeared, in contrast, to impart a small increase in resistance to inactivation by gamma-rays. When included in only one component, uvs-6 increased the sensitivity of homokaryotic uvs-6 conidia but had no affect on the other two conidial fractions. When included in both components, uvs-6 resulted in exponential inactivation curves and at the 30 krad exposure, 100-fold decreases in survival for all three conidial fractions.


Indications, contraindications and complications of laparoscopy. The primary use of laparoscopy is as a surgical tool, with sterilizations being the overwhelming indication. The laparoscope is used less frequently as a non-surgical tool, with the major indication being for diagnosing infertility and/or amenorrhea, and for evaluation of obscure pelvic pain. There would seem to be several indications for laparoscopy that have been neglected, these being in confirming the diagnosis of acute pelvic inflammatory disease; in the evaluation of malignancies and abdominal-pelvic trauma; and the surgical treatment of pelvic pain. Lapar-The majority of these contraindications are relative, and depend soley on the laparoscopist's ability and his clinical judgment. The problems of hernias seem to have been over-emphasized. The laparoscopist should be aware of potential problems with umbilical hernia, and he probably can ignore hiatal hernias except when they are large and quite symptomatic. However, generalized abdominal peritonitis, significant hemoperitoneum with intestinal obstruction are felt by most authors to be absolute contraindications. The most frequent complications of laparoscopy involve the physoperitoneum. Except for cardiac arrest the most serious complications involve electrical burns to small bowel.


[Postmortem fibrinolysis and its biological characteristics]. Fibrinolysic cadaveric blood studied after a rapid death (strangulation asphyxia, traumatic and sudden death) contained no plasmin--an enzyme of fibrin destruction, and no plasminogen--its inactive precursor. In the absence of plasmin and plasminogen of interest is a high content of plasminogen activator. An increase in the content of the activator in the fibrinolysic blood exists along with a marked proactivator level. A relationshp between the concentration of the activator and the proactivator in this blood indirectly pointed to the intravascular (from the pre-existing blood proactivator of the living), and not the tissue origin of the activator. Activation of plasma proactivator is caused by the entrance of tissue lysokinases into the circulation.


Mucopolysaccharides associated with nuclei of cultured mammalian cells. Mucopolysaccharides have been isolated, fractionated, and characterized from the nuclei of cultured B16 mouse melanoma cells grown in the presence of (3-H)-glucosamine and (35-S)sulfate. Digestion of the nuclei with DNase followed by Pronase gave a mixture of complex carbohydrates from which the mucopolysaccharides were isolated by precipitation with cetylpyridinium chloride. After fractionation by differential salt extraction and chromatography on controlled pore glass bead columns, the components were identified by chemical and enzymatic methods. The major polysaccharide components were a family of high-molecular-weight chondroitin sulfates with different degrees of sulfation; a minor component has been characterized as heparan sulfate.


The prone chest film.

The prone chest film. Frontal chest radiographs in the prone position enhanced visualization of the posterior lung base in the presence of pleural effusions. Though the lateral decubitus view often displaced fluid adequately, in several cases only the prone film revealed basilar lung pathology; this was due to gravitational shift of the fluid away from this region as well as improved aeration of the lower lobe in the prone position, as shown by horizontal-beam  lateral views. The prone chest film is recommended for evaluation of the lung base when obscured by fluid.


[Streptozotocin diabetes in a miniature pig (author's transl)]. The intravenous application of 60 mg streptozotocin/kg body weight to a Hanford miniature pig 8 days after a first dose of 30 mg/kg led to a diabetes with loss of insulin response to glucose, hyperglycaemia, glucosuria and a considerable increase of triglycerides and cholesterol. The application of 40 mg streptozotocin/kg to a second pig had no effect.


Filtration characteristics of dacron wool (Swank) blood transfusion filters. Stored human blood of varying age was passed through standard commercial blood transfusion filters (pore size, 170 mu) and Swank Dacron wool blood transfusion filters (pore size, 20mu). Passage through the Dacron filter resulted in a marked decrease in SFP and an increase in filter weight indicating removal of microaggregates which have been implicated as a cause of pulmonary insufficiency. The commercial filter tested did not appear to be effective in removing these harmful aggregates. On the basis of this research it is concluded that the Swank Dacron wool blood transfusion filter will prevent pulmonary microemboli during transfusion. Because aggregate removal appeared to be quantitative, it is recommended that no more than four units of blood be passed through each Dacron wool filter.


A 3-year survey of a rural orthopaedic clinic. The experiences of a team beginning and running a rural orthopaedic clinic are described. Suggestions for the running of such a clinic are put forward.


Predictive value of mixed leukocyte culture testing on allograft survival in unrelated dogs. This study evaluated the mixed leukocyte culture (MLC) technique to quantitate differences in kidney allograft survival between unrelated animals. Each of five beagles received two heterotopic kidney allografts, one from a mongrel dog showing high MLC stimulation and one from a second mongrel dog showing a low MLC stimulation. Low or high stimulators were defined by the amount of radioactive thymidine incorporated by lymphocytes when the recipient was tested against a number of potential donors in one way MLC TESTS. Low and high stimulators usually differed by a factor of up to ten in the stimulation index. Recipients were given a standard treatment of asathioprine and prednisone. No special treatment was given for rejection episodes. Creatine clearances and periodic biopsies were used to determine rejection. Creatine clearances and biopsies showed the high stimulating kidneys to be rejected completely by the end of 2 weeks in all animals. All of the low stimulating kidneys maintained normal function for 3 weeks and then developed progressive deterioration and rejection over the next 3 to 6 weeks. These studies suggest that quantitation of the MLC may be predictive of kidney allograft survival in unrelated dogs.


The effects of 5-fluorouracil and 5-fluorodeoxyuridine used alone and in combination with normal nucleic acid precursors on development of mice in lines selected for low and high expression of Strong's luxoid gene. Pregnant female mice selected for plus- and minus-modifying genes of the limb-skeleton effects of Strong's luxoid gene (1st) were injected ip on day 10 of gestation with 10 or 20 mg/kg 5-fluorouracil (5-FU) or 5-fluorodeoxyuridine (5-FUdR). The incidence and degree of expression of the effects (polydactyly, ectrodactyly, radial hemimelia, tibial hemimelia, pelvic girdle shifts, deformed caudal vertebrae) varied according to the teratogen and dose level. The addition of an equimolecular amount of thymidine, thymine, or uracil greatly potentiated the effects of 20 mg/kg 5-FU, usually resulting in death of the embryos. The addition of an equimolecular amount of thymidine but not of the embryos. The addition of an equimolecular amount of thymidine but not of thymine or uracil partly protected against the effects of 5-FUdR. Both teratogens increased the expression of ist on the limb skeleton. The interactions of the teratogens with the major gene were inhibited by minus- and promoted by plus-modifying genes of 1st. The effects of the teratogenic treatments may be mediated by cell death.


Fetal cardiovascular and other defects induced by thalidomide in cats. Timed pregnancies were induced in cats by synchronizing gonadotropin-stimulated estrus and ovulation with natural mating. The cats were given daily oral doses of thalidomide as follows: 10 mg/kg on days 10-20 of pregnancy, 240 mg/kg on days 10-14, and 120, 240, and 480 mg/kg on days 15-17 or 18-20 of pregnancy. Fetuses were delivered by cesarean section on day 44 or 45 or earlier if threatened abortion was considered imminent. A wide variety of cardiovascular anomalies was observed on gross and microscopic examination including the following ventricular septal defect, right atrial distension primarily involving the coronary sinus, malpositioned great vessels, and narrowed left ventricular chamber with hypertrophied walls. The overall incidence of these anomalies appeared related to dose and treatment period.


Molar concentrations of fibrinolytic components, especially free fibrinolysin, in vivo. The levels of fibrinogen and of profibrinolysin (plasminogen) in urokinase-treated plasma as a function of time of incubation were measured. The profibrinolysin concentration was estimated through its complete conversion to fibrinolysin and the inhibition of the enzyme by crystalline soybean trypsin inhibitor. The dissociation constant of the FL-STI complex was determined to be 7 times 10-9 M. The average concentration of profibrinolysin in normal human citrated plasma was found to be 8 times 10-7 M. From the decrease of fibrinogen with time in the urokinase-treated plasma, the free fibrinolysin was calculated. Free fibrinolysin in normal human blood in vivo was estimated from the half-life of fibrinogen and other data obtained in this study to be present at a concentration of 1.7 times 10-10 M. The plasmakinase activity in vivo, expressed as urokinase molarity, is also about 2 times 10-10 M.


Plasmin inhibition of thrombin-induced platelet aggregation. The effects of plasmin treatment upon washed human platelets were studied in an attempt to elucidate the mechanisms underlying thrombin-induced platelet aggregation. At calcium concentrations of 10-20 muM, PLASMIN (0.2 CTA U/ml) inhibited thrombin-induced aggregation almost completely, but did not diminish the thrombin-induced release of adenine nucleotides, 5-hydroxytryptamine, or calcium. Increasing the calcium concentration partially antagonized plasmin's inhibition of aggregation. Studies utilizing calcium chelators and the Kunitz soybean trypsin inhibitor (SBTI) as a plasmin inhibitor indicated that in order to achieve maximal block of aggregation, plasmin must act upon a substrate made fully available only after an initial thrombin-platelet interaction has taken place. Moreover, the time course of this inhibition parallels the time course of the thrombin-induced release reaction. Plasmin inhibition of aggregation could not be mimicked by exposing the platelets to proteolytic digests of fibrinogen at concentrations as high as 17% total platelet protein. Nor could inhibitory activity be recovered from supernatants of plasmin-treated platelets, upon centrifugation and treatment with SBTI. With the use of a "cold initiation" technique, the release by thrombin of 46.7 plus or minus 6.7 (mean plus or minus SEM) mu-g of fibrinogen immunological equivalents per mg platelet protein could be demonstrated. Platelets in which thrombin-induced aggregation was abolished by plasmin treatment (and the plasmin subsequently inactivated by STBI) aggregated normally upon addition of as little as 10 mu-g human plasma fibrinogen per mg platelet protein. It is concluded that plasmin inhibition of aggregation most likely results from its attack upon a protein that is released or becomes fully available subsequent to interaction of thrombin with a platelet receptor mediating release. The results of this study are consistent with a cofactor role for fibrinogen in the aggregation of human platelets by thrombin.


[Transposition of great vessels].

[Transposition of great vessels]. Venous inflow stenoses either pulmonary or systemic represents a major complication after Mustard's operation. In order to eliminate such stenoses two modified methods of atrial in version have been developed using no or a minimum of foreign material. The indication for each method is different. Both methods have the incorporation of the remaining atrial septum and of a part of the left atrial wall into the new septum in common. The midportion is completed using a small Dacron patch in cases with fragile atrial tissue (method I) or a flap of the rightatrial wall in cases with normal tissue (method II). The incorporation of growing tissue into the new atrial septum minimizes the risk of secondary stenoses, due to shrinkage or excessive apposition of fibrous pannus.


Inhibition of the mixed leucocyte culture reaction by plasma from renal transplatn recipients. Plasma samples from 28 renal allograft recipients were studied for the presence of alloantibodies blocking MLC response. MLC blocking antibodies were found in plasma samples obtained from 19 recipients, six with rejected allograft and 13 with long-term surviving transplant. In many instances, these antibodies were found in the absence of detectable lymphocytotoxic antibodies. The MLC blocking activity resides in IgG fractions of plasma samples and was decreased only after absorption with lymphocytes, no effect on blocking was observed by absorption with platelets and fibroblasts.


Further miniaturization and automation of in vitro lymphocyte cultures. A microtechnique (10 mul culture volume) for in vitro lymphocyte cultures is described, which, compared with current miniaturized techniques, permits a four to 20-fold reduction of both medium volume and cell numbers. Furthermore, two alternative procedures for harvesting and washing radioisotope labelled cells are described. The first, making use of a conventional harvesting device, collects cells on glass fibre filters in a two to three-fold shorter time than that needed for the current semi-automatic collectors. The second takes advantage of the possibility of washing and fixing the cells in their culture wells and allows processing of more cultures with fewer manual operations. Various technical aspects of the micro-culture system are described and discussed with special reference to automation problems.


Possible use of established cell lines for MLR locus typing. MLR histocompatibility typing was performed by in vitro stimulation of lymphocytes from various persons with fresh lymphocytes (irradiated) or with lymphoblostoid cell lines (mito-mycin-treated) derived from donors homozygous at the MLR locus. MLC responses to both types of stimulation correlated strongly suggesting that a) established lymphoblastoid line cells can express products of the MLR loc and b) fresh lymphocytes can be replaced by these long term lines in MLR typing. Two lines (LV-B and MS-B) derived from two MLR homozygous donors (lv and MS) were used throughout these experiments. It is possible that as additional lines are established from such MLR homozygous donors, a panel of typing cells could be created which would serve to standardize MLR typing and allow selection of unrelated but histocompatible donors for transplantation.


The use of viable frozen lymphocytes for studies in human tumor immunology. Viable frozen lymphocytes displayed activity in blastogenesis assays that was indistinguishable from freshly prepared lymphoid cells. Similarly, cytotoxic activity of lymphocytes against melanoma target cells from melanoma patients was only slightly affected by the freezing procedure. Frozen  lymphocytes provided a highly reproducible source of cells in these assays. The use of viable frozen peripheral blood lymphoid cells for the retrospective analysis of a cancer patient's immune response is described.


Islet transplantation in diabetic rats receiving corticosteroids. Pancreatic tissue fragments from neonatal inbred Lewis rats were distributed throughout the peritoneal cavities of isologous diabetic recipients receiving prednisolone on a schedule similar to that used in clinical transplantation beginning 9-44 days before transplantation. Plasma glucose levels in six of seven diabetic rats receiving prednislone returned to normal levels 3 weeks after transplantation. Plasma insulin levels in the animals with successful transplants reached, at 1 week after transplantation, levels measured in normal control rats, but never attained the high insulin levels observed in the normal control animals receiving steroids. As with glucose the insulin levels remained in the normal range, and the animals steadily gained weight. During i.v. glucose tolerance tests (IVGTTs) in the rats with successful transplants at 4, 11-14, and 26-29 weeks after transplantation, the glucose levels were similar to those observed during IVGTTs in the control animals before and during corticosteroid therapy. In contrast, insulin levels during the IVGTTs were lower in the rats with successful transplants than in the control rats with or without steroids; however, insulin release during the IVGTTs was rapid in all animals, both with successful transplants and controls. These observations demonstrate that islet tissue implanted in the peritoneal cavities of diabetic rats receiving immunosuppressive chronic corticosteroid therapy can respond with a rapid and sufficient release of insulin to reverse diabetes as documented by normal glucose levels under stimulated and nonstimulated conditions.


Immune injury from organ preservation. A potential cause of hyperacute rejection in human cadaver kidney transplantation. Two pairs of plasma-perfused human cadaver kidneys were rejected in a hyperacute manner by recipients who had not previously received a transplant. Crossmatches between recipient sera and donor lymphocytes were negative in all cases. A fifth kidney was plasma-perfused but not transplanted because the perfusate was shown to be cytotoxic to donor lymphocytes. IgM and complement, but not IgG, were demonstrated in these kidneys by immunofluorescent microscopy and confirmed by further immunological studies. The IgM was broadly reactive against multiple HL-A specificities and was present in 11 percent of sera from normal, healthy male donors. It appears from our studies that cytotoxic IgM may be present in homologous plasma and cause immune injury to the kidney during ex vivo pulsatile preservation. This may be responsible for some cases of otherwise unexplained accelerated allograft rejection.


[Comparative karyological study of 2 close species of blackflies of the genus Eusimulium]. The structure of polytene chromosomes in the salivary gland of two hard for distinguishing species Eusimulium montium Rubz. and E. gviletense alizadei Djaf. was studied. The comparative-karyologic analysis of the taxons showed many common features in their karyotypes: complete conjugation of the homologous chromosomes in the connection between the length and the arms of chromosomes. At the same time these species are well distinguished karyologically according to the two inversions of the homozigotic type in the chromosomes I and III, to the duplication in IIL, and to the deficiency of the telometric region in IIIL of E. montium. These readjustments are considered to be secondary which suggests E. gviletense alizadei to be phylogenetically older than E. montium.


Evolution of urinary flow rate with prostatectomy. Urinary flow rate was studied before and after transurethral prostatectomy in 53 patients. An average of 43.8 percent improvement in maximum flow rate after transurethral prostatectomy was observed in this series. Improvement of maximum flow rate occurred in 71 per cent of patients. Half of the cases in which flow rate did not improve were chronic prostatitis. Uroflowmetry cannot be utilized as a quality index of a transurethral prostatic resection as long as the degree of detrusor deterioration is not assessed. There is a relationship between the degree of preoperative symptoms and the degree of postoperative urinary flow improvement. The degree of preoperative symptoms and urinary flow deviation are not related which suggests that uroflowmetry adds to the assessment of symptoms in the determination of the degree of obstruction. Patients presenting difficulty of urination have a greater chance to demonstrate urinary flow improvement after surgery than patients complaining of urinary frequency.


[Effect of heparin on the Mg++-ATPase of the muscle mitochondria of adult and young rabbits]. ATPase activity from freshly prepared, "aged" and "heparin" mitochondria was studied in adult, newborn and 7 day old rabbits. Heparin, incubated with the mitochondria within 2-2.5 hrs at 20 degrees C, increased the activity of endogenous ATPase in the newborn rabbits. Activities of Mg-2++-ATPase and Mg-2+-DNP-ATPase were decreased under effect of heparin in both adult and new born rabbits. In 7 day old rabbits, to the contrary, increased activity of Mg-2+-ATPase and its activation by 2,4-DNP were observed.


[Incidence and etiology of omphaloceles (author's transl)]. Omphaloceles are congenital malformations with herniation of the abdominal viscera into the umbilical cord. The incidence in newborns is 1:4300, the sex ratio 1.24:1 in favour to boys. Children with this malformation are neither more frequent at the beginning nor at the end of the sibship; the average maternal age is not increased. In about 50% other different malformations are found. All those omphaloceles, which are found as an obligatory symptom in the EMG syndrome (Wiedemann-Beckwith syndrome) to be probably in most cases autosomal recessively inherited or which are seen within a chromosomal malformation syndrome are of etiologically known origin. Omphaloceles without other malformations are mainly sporadic, less frequent in sibs, possibly due to a polygenic or multifactorial mode of inheritance. Findings in population genetics and animal experiments suggest that exogenous factors, too, can be the cause of malformation syndromes with omphaloceles.


The contents of acid glycosaminoglycans in the aortic wall of newborns, small children, and older adults. The contents of acid glycosaminoglycans (aGAG) was determined in the inner and medial layers of aortae from 13 newborns, 9 small children (1-5 years of age), and 10 older adults (7th decade of life). The total aGAG contents was expressed in mug hexuronic acid/100 mg dry weight. Newborns and older adults showed similiar contents of aGAG in their aortae (275.5 plus or minus 27.9 and 313.4 plus or minus 46.7, respectively). Small children had significantly less aGAG in their aortae (202.1 plus or minus 21.5). The aGAG material from aortae was fractionated by a microcolumn chromatographic procedure on cetylpyridinium chloride cellulose. No significant differences of the aGAG fractions were found in aortae of newborns and children. In both groups of children the percentage of hyaluronic acid was significantly increased and dermatan sulfate was significantly decreased when compared with aortae from older adults. The increased contents of acid glycosaminoglycans (aGAG) in aortae of human newborns is possibly caused by the concentrated hormonal environment (especially estrogens) to which the fetus is exposed in utero.


[The primary extranodal manifestation of lympho- and reticulosarcoma of the stomach]. We report about a group of 11 patients with primary lympho- or reticulosarcoma of the stomach in a group of 94 patients with Non-Hodgkin-Lymphomas. There were 5 patients with reticulosarcoma of the stomach among 44 patients with reticularsarcoma (11 percent) and 6 patients with primary lymphosarcoma of the stomach among a total of 50 patients with lymphosarcoma (12 percent). In most instances the diagnosis was made during surgery. At the time of diagnosis only 4 of the 11 patients had Stage IE disease. All patients who received intensive chemotherapy are living. Patients who died during the observation period had reticulosarcomas. They had not received any chemotherapy or treatment was discontinued early for varying reasons. Results are discussed in view of the diagnosis and treatment, suggestions for therapy are made.


Reproducibility of survival time in L 5222 rat leukaemia and its implications for chemotherapeutic tests. The reproducibility of survival time in the L 5222 leukemia in rats was tested by transfer of varying numbers of untreated and X-irradiated cells. A linear relationship between log cell dose and survival time was established for the range of unirradiated cell doses between 10-8 to "10-0", resulting in survival times from 6 to 16 days, with very little variation between individual animals of each group. This narrow deviation of survival time makes it possible to use mean survival time as a measuring parameter instead of the cell dose required to kill 50 percent of recipient animals. From the longer survival times observed after transfer of a given number of X-irradiated cells, the number of viable cells transferred in the inoculum could be calculated and thus the degree of cell death due to X-radiation. Agains a correlation between log cell dose and survival time was found but host survival was prolonged when small numbers (similar to 10-2) of viable irradiated cells were transferred. It is suggested that this is not due to a change in proliferation kinetics but rather to host factors, such as an immunological reaction. Thus the L 5222 leukaemia seems to be a good model in its reproducibility of survival time, and may have some similarity to human acute leukaemia when low numbers of treated cells are concerned.


[Rat urinary metabolites of (2-hydroxybutyl)-n-butylnitrosamine]. The chemical structure of 3 urinary metabolites of 2-hydroxy-di-n-butylnitrosamine has been identified by mass spectrometry. The results indicate that oxidation and formation of metabolites with shorter chain length may be explained by alpha-oxidation and subsequent degradation. This is an alternative to the beta-oxidation hypothesis of Krüger (1971).


[Electronmicroscopic studies on solid mammary carcinomas in bitches]. The electronmicroscopic findings of nine solid mammary carcinomas in bitches are described as well as the observations on tumour cells within lymphatic vessels. In the primary tumours basement membranes surround the cords of tumour cell. In five cases, particularly in two, there are cells with long intracytoplasmic filaments measuring about 65 A thick. The filaments are arranged along the axis of the cells and show focal densities. These cells are presumed to be myoepithelial tumour cells. In eight of the nine tumours there are also carcinoma cells with short single or bundled intracytoplasmic filaments of approximately the same thickness (65 A). These however show no specific order, they are perinuclear or lie in between the organelles. They are called tonofibrillae. Viruses or virus-like particles have not been found.


Sensitivity tests of tumors to cytostatic agents. I. Comparative investigations on transplanted tumors in vivo and in vitro. With series of transplanted tumors, the activities of different cytostatic agents which directly influence the metabolism of nucleic acids (Actinomycin D, adriamycin, daunomycin, 5-fluorouracil, procarbazine, trenimon) was measured by determining 3-H-uridine incorporation in short-term (3hrs) incubations of  tumor cell suspensions. Data obtained could be used to predict the response of each tumor to particular cytostatic agents in vivo. The activities of the cytostatic agents as determined using  long-term tissue cultures (time of exposure of tumor cells to cytostatic agent 48 hrs were comparable to those obtained with the short-term test.  In long-term cultures, determination of cell numbers gave results similar to those obtained by morphological evaluation. In SHORt-term test, differing sensitivities of tumors to cytostatics could be detected.


Sensitivity tests of tumors to cytostatic agents. II. Investigation on human tumors. Certain substances which influenced nucleic acid metabolism were found to have about the same cytostatic activity on human cells when measured in tissue culture experiments (cell numbers) or in short-term cultures (3-H-uridine incorporation in cell suspensions). By treatment with a dose of cytostatics corresponding to 10 times therapeutic dose, chemosensitive tumors can be distinguished from non-responsive tumors. By using this in vitro test system to investigate the sensitivities of 100 human tumors, it is shown that 28 of these tumors were responsive to adriamycin, daunomycin and Actinomycin D. Good agreement was observed between these in vitro results and the literature data on clinical therapy using these particular substances.


[Result of a user inquiry for the improvement of hand propelled wheelchairs (author's transl)]. The contribution describes the results of an inquiry of handicapped wheelchair drivers which have been compiled by evaluating 3482 questionnaires with 268,144 informations using an IBM 1130 computer. Divided into "rim" and "lever" propulsion the driving quality, the manoeuvrability, the seat comfort, problems of driving mechanism and the transportation suitability as well as the wear of the vehicles are analysed. Finally some recommendations for the design of future wheelchair models are given on the basis of the observed drawbacks.


Foetal and maternal plasma levels of steroid sulphates in human pregnancy at term. Umbilical cord arterial and venous, and maternal peripheral vein blood samples were collected during 20 deliveries after uncomplicated pregnancies. Using a gas chromatographic method, oestriol, oestriol sulphate, progesterone and a number of neutral steroid mono- and disulphates were quantitated in the cord plasma samples and a number of neutral steroid mono- and disulphates in the maternal plasma samples.


Atypical myoclonus body epilepsy (adult variant). A young woman, with negative family history, committed suicide after 6 years of myoclonus and mental deterioration. The neurons contained atypical myoclonus bodies which differed from  the classical type in distribution, histochemistry and light and electron microscopic appearance.


Staphylococcal bacteremia and hexachlorophene bathing. Epidemic in a newborn nursery. An outbreak of staphylococcal bacteremia in healthy, full-term neonates occurred in the newborn nursery at a large county hospital not employing prophylactic hexachlorophene bathing. Seven infants had staphylococcal bacteremia and one had omphalitis. Two of the three isolates obtained for phage typing were type 86, and the other was 3c/71. Staphylococcal colonization rate in the nursery was 64% when the outbreak was recognized; 86% of these isolates were type 86. No predominant phage type was isolated from nursery personnel. The outbreak followed a six-month preliminary study that showed a rise in staphylococcal colonization rate from 2.2% with hexachlorophene bathing to 67% with a nonhexachlorophene-containing preparation. In a community survey of infants born during the two months prior to the epidemic, seven of eight babies with lesions were infected with Staphylococcus aureus type 86.


Extraglomerular immunoglobulin deposits in human nephritis. In order to evaluate the incidence of extraglomerular immunoglobulin deposits and to correlate their presence with histopathologic abnormalities, we performed both prospective and retrospective immunofluorescence studies of renal biopsy specimens. Of 200 diagnostic biopsy specimens examined prospectively, 21 had extraglomerular deposits, 19 in association with presumed immunologically-mediated glomerulonephritis. Nine had linear immunoglobulin deposits on tubular basement membrane (antitubular basement membrane antibodies), in one case coexisting with granular deposits, and 13 had granular deposits on tubular basement membrane, in peritubular capillaries and/or arteries, or in tubular cytoplasm (probably immune complexes). Linear deposits on tubular basement membrane were usually associated with antiglomerular basement membrane nephritis or methicillin-associated interstitial nephritis; granular extraflomerular deposits were seen primarily in systemic lupus erythematosus, cryoglobulinemia or membranoproliferative glomerulonephritis.. The incidence of immunoglobulin deposits was high in the three groups of patients examined retrospectively being present in 37 or 47 patients with antiglomerular basement membrane mephritis, 22 or 32 patients with systemic lupus erythematosus of 24 of 130 renal allograft recipients. The contribution of antitubular basement membrane antibodies to renal damage was difficult to assess, although studies in experimental animals attest to their pathogenicity. The occurrence of granular extraglomerular deposits appeared to correlate roughly with the degree of tubulointerstitial injury in the patients with systemic lupus erythematosus.


Radiosensitivity of sensitized and nonsensitized human lymphocytes evaluated in vitro. The purpose of this study is to compare the relative radiosensitivity of sensitized and nonsensitized lymphocytes utilizing an in vitro test system. The test system employed is the combined mixed leukocyte reaction and cell-mediated lympholysis assays (MLC-CML). In this system, sensitized T lymphocytes of human origin are much more resistant to radiation-induced injury than are nonsensitized lymphocytes. The results support similar observation in vivo with antigen-activated lymphocytes and are interpreted in one of two ways: a) sensitized T cell, rendered nonviable by radiation, are still able to exert lytic activity, perhaps via the release of cytotoxic factors; and b) sensitized T cells are inherently radioresistant, perhaps due to the hypermetabolic state of such cells, which may serve to activate repair mechanisms.


Ultrastructural features of degenerated cardiac muscle cells in patients with cardiac hypertrophy. Degenerated cardiac muscle cells were present in hypertrophied ventricular muscle obtained at operation from 12 (38%) of 32 patients with asymmetric septal hypertrophy (hypertrophic cardiomyopathy) or aortic valvular disease. Degenerated cells demonstrated a wide variety of ultrastructural alterations. Mildly altered cells were normal-sized or hypertrophied and showed focal changes, including preferential loss of thick (myosin) filaments, streaming and clumping of Z band material, and proliferation of the tubules of sarcoplasmic reticulum. Moderately and severely degenerated cells were normal-sized or atrophic and showed additional changes, including extensive myofibrillar lysis and loss of T tubules. The appearance of the most severely degenerated cells usually reflected the cytoplasmic organelle (sarcoplasmic reticulum, glycogen, or mitochondria) which underwent proliferation and filled the myofibril-free areas of these cells. Moderately and severely degenerated cells were present in areas of fibrosis, had thickened basement membranes, and had lost their intercellular connections. These observations suggest that degenerated cardiac muscle cells have poor contractile function and may be responsible for impaired cardiac performance in some patients with chronic ventricular hypertrophy.


Idiopathic chylopericardium: 131-I-triolein scan for noninvasive diagnosis. We report idiopathic chylopericardium in a physically active, asymptomatic 29-year-old man. Preoperative diagnosis was made by external cardiac imaging after oral administration of 131-I-labeled triolein. To our knowledge this is the first report of preoperative noninvasive diagnosis of chylopericardium. After open drainage and pericardial biopsy, the patient is asymptomatic without recurrence after 8 months.


Potentiation of the toxicity of mithramycin by bacterial lipopolysaccharide. The lethality for BALB/c mice of 1,3-bis(2-chloroethyl)-1-nitrosourea, cytosine arabinoside, 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea, hydroxyurea, mithramycin, a polymyxin-like antibiotic (SQ 21,286), polyadenylic polyuridylic acid, procarbazine, 5-[3,3-bis(2-chloroethyl)-1-triazeno]-imidazole-4-carboxamide (TIC-mustard) or uracil arabinoside administered in combination with Escherichia coli lipopolysaccharide (LPS) was measured. Simultaneously administered mithramycin and LPS or TIC-mustard administered 24 h after LPS synergistically killed mice. Concanavalin A potentiated the lethality of TIC-mustard but not of eight other drugs tested. Pretreatment of mice with LPS or lipid A complexed to concanavalin A rendered mice resistant to the lethal action of LPS alone or combinations of LPS and mithramycin. Mithramycin-treated mice were killed by minute amounts of LPS. Mice sensitized to LPS by mithramycin were used to detect endotoxic activity in biological materials, such as commercially available enzymes, and in a complex of lipid A with concanavalin A.


Sebum: analysis by infrared spectroscopy. II. The suppression of fatty acids by systemically administered antibiotics. Various tetracyclines and erythromycins have been used rather empirically for the systemic treatment of acne vulgaris for more than a decade. It has been impossible to accurately evaluate clinically the numerous derivatives and to compare their effectiveness with other antibiotics such as clindamycin. As a substitute for this approach, this present study ranks these antibiotics, when given orally, as to their effectivensss in suppressing the fatty acids in sebum in normal volunteers. As a group, the tetracyclines were more effective than the erythromycins in decreasing the fatty acids, but clindamycin was significantly more potent than either.


The relationship of axillary venous thrombosis to the thoracic outlet compression syndrome. In a relatively small personal series (41 patients) of first rib resections for thoracic outlet compression, I have encountered 8 patients in whom venous obstruction at the thoracic outlet was the major component. Four of the 8 had typical "stress" thrombosis of the axillary vein. Angiography demonstrated a tight thoracic outlet to be the underlying cause of the venous thrombosis. In the 4 patients with venous compression but without thrombosis, angiograms suggested that each was a candidate for axillary venous thrombosis unless the underlying thoracic outlet compression syndrome was relieved. Although thoracic outlet compression has been documented as a cause of "stress thrombosis" of the axillary vein, the relationship of the two conditions apparently is often overlooked, even by experienced thoracic surgeons. Representative case histories and angiograms are presented to demonstrate the close relationship of thoracic outlet compression and axillary venous thrombosis.


Total anomalous pulmonary venous return. Review of 125 patients treated surgically. One hundred twenty-five patients undergoing surgical correction of total anomalous pulmonary venous return were studied. The overall mortality was 37% and was related to age at the time operation was required. Mortality was 57% during the first year of life, 29% in patients between 13 and 24 months, and 15% in those between 2 and 10 years; no deaths occurred in those over 10 years. Mortality was highest in patients with infracardiac lesions (62%), and lowest in those with cardiac defects (30%). The major cause of death was pulmonary edema, and survival was closely related to the degree of increased pulmonary vascular resistance. Surgical treatment should be delayed until at least 6 months of age, but the development of congestive heart failure may necessitate earlier operation.


Myocardial performance of disabled patients in a rehabilitation program. Thirty-two patients with heart disease in addition to another disability were monitored continuously with the ten-hour electrocardiographic (ECG) tape. Eighteen of the 32 patients manifested abnormalities of rate, rhythm or ST-segment depression. Patients with arrhythmia eventually did better than those with other ECG abnormalities because drug therapy was successful in abolishing the abnormal rhythm in most cases. Those patients with excessive tachycardia did poorly as a group in so far as their rehabilitation program was concerned. This technique of continuous ECG monitoring may be helpful in assessing myocardial performance in these patients and in defining the prescription for rehabilitation therapy.


Measles rash. I. Light and electron microscopic study of skin eruptions. Measles skin lesions were studied by light and electron microscopy. In the epidermis multinucleated giant cells were observed just beneath the hypertrophic horney layer at the maximum stage of rash; they were believed to result by an abnormal process of hyper- or parakeratosis. Neither typical inclusions nor viral nucleocapsids could be detected in any part of the epidermal layer. Most characteristical changes were dermal edema and spongiosis with mononuclear cell infiltration as well as the detection of measle virus-like microtubular structures (nucleocapsids) in the endothelium of dermal capillaries. Is is assumed that measles exanthema is a manifestation of an Arthus reaction elicited by viral antigen in the endothelium of dermal capillaries.


Zinc distribution in mouse brain subsequent to hippocampal lesions. The role of zinc in central nervous system metabolism remains obscure but it has been shown in animals that diphenylthiocarbazone (Dithizone) selectively colors intravitally the hippocampus and adnexa, parts affected in human temporal lobe epilepsy. To examine the effect of experimental cerebral lesions on zinc storage in that region, 80 mice were divided into two operative and two control groups. The operative groups had unilateral lesions placed in the hippocampus or frontal lobe. One control group and both lesion groups were injected subcutaneously with zinc lactate daily for ten days before intravenous injection of diphenylthiocarbazone. The other control group was used to determine if zinc storage occurs as a result of its increased systemic availability. Hippocampal lesion mice showed more extensive staining of lethal amygdala and associated cortex on the side of the lesion. Frontal lesions remained unstained and frontal lesions did not alter the staining of hippocampus and related parts. Lack of difference between controls shows that greater availability of systemic zinc does not increase its content in the hippocampal-lateral amygdalar region. Increased zinc uptake in this instance appears to be a local phenomenon.


Acrodermatitis enteropathica. Corneal involvement with histochemical and electron micrographic studies. A case of acrodermatitis enteropathica occurred with corneal changes consisting of linear subepithelial corneal opacities. From the results of histochemical and electronmicrographic studies on a biopsy specimen of the corneal lesion, no definite conclusion may be drawn as to whether the corneal changes are a manifestation of acrodermatitis enterophathica or a result of iodochlorhydroxyquin therapy.


Studies on a plasma cardiac glycoside assay based upon displacement of 3-H-ouabain from Na+-K+-ATPase. We tested an assay system introduced for plasma glycoside measurements, basing on the displacement of 3-H-ouabain from Na+-K+-ATPase by unlabeled glycoside. ATPase preparations from hog, cat and guinea pig were used. Displacements were performed using 20 cardiac glycosides, genins and derivatives with different cardiac activity. Most of the glycosides and derivatives do not induce a continuous 3-H-ouabain displacement from the ATPase, but a very steep increase of unbound 3-H-ouabain between 10-minus 7 and 2 times 10-minus 7 M. Therefore this assay system shows a satisfactory discrimination only in a short concentration range. This behavior and a relatively low sensitivity make the ATPase displacement assay problematic for clinical and pharmacokinetical plasma glycoside measurements.


Evaluation of RX 72601 as an anti-curare agent. RX 72601, a new and potent anticholinesterase, has been evaluated for its ability to reverse the neuromuscular blockade induced by non-depolarizing muscle relaxants. In rats RX 72601 10 mug/kg i.v. proved effective in reversing the effects of alcuronium gallamine, pancuronium or tubocurarine and the drug exhibited a wide margin of safety. RX 72601 proved equally effective in reversing the effects of tubocurarine in both cats and baboons. In dogs and cats, effective anti-curare doses had little action on the cardiovascular or respiratory systems. Overall the results obtained indicate that RX 72601 may be a safe antagonist of non-depolarizing muscle relaxants and that it will be possible to use RX 72601 without prior administration of atropine.


Morphological and biochemical defects in muscles of human carriers of the malignant hyperthermia syndrome. This communication presents evidence in support of a neuropathic basis for the myopathy associated with malignant hyperpyrexia (MH). Muscle from MH susceptible individuals showed a reduced calcium uptake by the sarcoplasmic reticulum. There was a reduced concentration of phosphocreatine and ATP and an increased concentration of glucose-6-phosphate in these muscle samples.


Human cardiac myosin ATPase and light subunits. A comparative study. Myosin was extracted from normal human hearts (autopsy material) and compared to that of pig heart and rabbit white skeletal muscle. Myosin light subunits were isolated by a preparative urea gel electrophoresis. These subunits were shown by urea and sodium dodecylsulfate gel electrophoresis to be only slightly affected by the time lapse between death and the beginning of myosin extraction. This was also true for myosin ATPases. The Ca-2+-activated ATPases of pig and human heart myosins have the same apparent Km and V, whereas white skeletal muscle myosin ATPase has the same Km with a higher V. Human myosin light subunits, when compared to those of pig heart possess: (i) different molecular weights: 27 999 and 18 000 datlons for pig heart, and 25 000 and 19 000 daltons for human heart. (ii) for both the light chains, different ultraviolet spectra and a higher helical content for the subunit molecular weight 25 000. (iii) a different composition for several amino acids (Tyr, Pro, Lys). A third light subunit (molecular weight 15 000) was occasionally seen in human as well as pig heart myosin. It concentration varied inversely with that of the subunit molecular weight 27 000-25 000, and so was probably a degradation product of the heaviest subunit.


Amino acid sequence of the carboxy-terminal cyanogen bromide peptide of the human fibrinogen beta-chain: homology with the corresponding gamma-chain peptide and presence in fragment D. The carboxy-terminal cyanogen bromide fragment of the human fibrinogen beta-chain has been isolated and its structure determined. It is a nonapeptide with the sequence Lys-Ile-Arg-Pro-Phe-Phe-Pro-Gln-Gln and is homologous with a portion of the carboxy-terminal cyanogen bromide fragment of the gamma-chain. The peptide has also been isolated in full yield from cyanogen bromide digests of the plasmin-derived fragment D, indicating that the carboxy-terminal region of the beta-chain is resistant to plasmin digestion. In contrast, a small portion of the corresponding gamma-chain carboxy-terminal region was missing in the same fragment D.


Gel electrophoretic identity of the (Na+ + Mg-2+)- and (Na+ + Ca-2+)-stimulated phosphorylations of rat brain ATPase. The classical E2-P intermediate of (Na+ + K+)-ATPase dephosphorylates readily in the presence of K+ and is not affected by the addition of ADP. To determine the significance in the reaction cycle of (Na+ + K+)-ATPase of kinetically atypical phosphorylations of rat brain (Na+ + K+)-ATPase we compared these phosphorylated components with the classical E2-P intermediate of this enzyme by gel electrophoresis. When rat brain (Na+ + K+)-ATPase was phosphorylated in the presence of high concentrations of Na+ a proportion of the phosphorylated material formed was sensitive to ADP but resistant to K+. Similarly, if phosphorylation was carried out in the presence of Na+ and Ca-2+ up to 300 pmol/mg protein of a K+ -resistant, ADP-sensitive material were formed. If phosphorylation was from [gamma-32-P]CTP up to 800 pmol-32-P/mg protein of an ADP-resistant, K+ -sensitive phosphorylated material were formed. On gel electrophoresis these phosphorylated materials co-migrated with authentic Na+ -stimulated, K+ -sensitive, E2-P-phosphorylated intermediate of (Na+ + K+)-ATPase, supporting suggestions that they represent phosphorylated intermediates in the reaction sequence of this enzyme.


Showdomycin, a nucleotide-site-directed inhibitor of (Na+ + K+)-ATPase. Showdomycin [2-(beta-D-ribofuranosyl)maleimide] is a nucleoside antibiotic containing a maleimide ring and which is structurally related to uridine. Showdomycin inhibited rat brain (Na+ + K+)-ATPase irreversibly by an apparently bimolecular reaction with a rate constant of about 11.01-mol- minus 1-min- minus 1. Micromolar concentrations of ATP protected against this inhibition but uridine triphosphate or uridine were much less effective. In the presence of K+, 100 MUM ATP was unable to protect against inhibition by showdomycin. These observations show that showdomycin inhibits (Na+ + K+)-ATPase by reacting with a specific chemical group or groups at the nucleotide-binding site on this enzyme. Inhibition by showdomycin appears to be more selective for this site than that due to tetrathionate or N-ethylmaleimide. Since tetrathionate is a specific reactant for sulfhydryl groups it appears likely that the reactive groups are sulfhydryl groups. The data thus show that showdomycin is a relatively selective nucleotide-site-directed inhibitor of (Na+ + K+)-ATPase and inhibiton is likely due to the reaction of showdomycin with sulfhydryl group(s) at the nucleotide-binding site on this enzyme.


Action of cytochalasin A, a sulfhydryl-reactive agent, on sugar metabolism and membrane-bound adenosine triphosphatase of yeast. Cytochalasin A at 10-20 mug/ml inhibits growth and sugar uptake by Saccharomyces strain 1016. The effects of cytochalasin A in intact cells were completely prevented when 1 mM cysteine or dithiothreitol was added along with cytochalasin A, but were not eliminated by thiols added after inhibition had occurred. Purified yeast hexokinase, glucose-6-P dehydrogenase, phosphofructokinase and aldolase were not sensitive to cytochalasin A (20 mug/ml). Glyceraldehyde-3-P dehydrogenase was strongly inhibited by cytochalasin A (5 mug/ml); activity was promptly restored by thiols. Anaerobic glycolysis was inhibited by cytochalasin A or by iodoacetate; unlike iodoacetate, cytochalasin A did not cause accumulation of sugar phosphates. In contrast, cytochalasin A, but not iodoacetate, inhibited isolated membrane-bound ATPases. Cytochalasin A is a sulfhydryl-reactive agent and has membrane-related effects (adenosine triphosphatase) which may well be the basis of its interference with energy-dependent uptake of solutes.


Isolation and characterization of two types of sarcoplasmic reticulum vesicles. A purified preparation of sarcoplasmic reticulum from rabbit skeletal muscle has been found to consist of a heterogeneous population of vesicles. Isopycnic centrifugation was used to obtain "light" and "heavy" vesicles from the upper and lower ends of a 25 to 45% (w/w) linear sucrose gradient. Each fraction accounted for about 10 to 15% of the total vesicles. The remainder of the vesicles were of intermediate density and banded between the light and heavy fraction. Light vesicles were composed of about equal amounts of phospholipid and Ca-2+ pump protein which contained approx. 90% of the protein. Heavy vesicles contained in addition to the Ca-2+ pump protein (55-65% of the protein) two other major protein components, the Ca-2+ binding and M55 proteins which accounted for 20-25 and 5-7% of the protein of these vesicles, respectively. The sarcoplasmic reticulum subfractions had 32-P-labelled phosphoenzyme levels proportional to their Ca-2+ pump protein content and contained similar Ca-2+-stimulated ATPase activities. They were capable of accumulating Ca-2+ in the presence of ATP and of releasing the accumulated Ca-2+ when placed into a medium with a low Ca-2+ concentration. The vesicles differed significantly in that heavy vesicles had a greater number of non-specific Ca-2+ binding sites than light vesicles (approx. 220 vs 75 nmol of bound Ca-2+ per mg protein), in accordance with their high content of Ca-2+ binding protein. Electron dense material could be seen within the compartment of heavy but not light vesicles. Removal of Ca-2+ binding and M55 proteins from heavy vesicles resulted in empty membranous structures consisting mainly of Ca-2+ pump protein and phospholipid. Electron micrographs of sections of muscle showed dense material in terminal cisternae but not in longitudinal sections of sarcoplasmic reticulum. These experiments are consistent with the interpretation that (1) the electron dense material inside heavy vesicles may be referable to Ca-2+ binding and/or M55 proteins, and that (2) light and heavy vesicles may be derived from the longitudinal sections and terminal cisternae of sarcoplasmic reticulum, respectively.


Membrane reconstitution in chl-r mutants of Escherichia coli K 12. VII. Purification of the soluble ATPase of supernatant extracts and kinetics of incorporation into reconstituted particles. Membrane-bound ATPase (EC 3.6.1.3) of Escherichia coli K 12 is released in a soluble form by the mechanical treatments applied to the cells in order to break them. The purification of the soluble enzyme is described. The purified protein gives a single band in 7.5% polyacrylamide gel electrophoresis. The molecular weight is estimated to be 350 000. The enzyme is cold-labile, Mg-2+ dependent, insensitive to inhibition by N, N'-dicyclohexylcarbodiimide and specific for ATP and ADP. Membranes depleted of their ATPase activity by dilution in a buffer of low ionic strength and without Mg-2+ are able to incorporate the purified ATPase only in the presence of 2-6 mM Mg-2+. ATPase binds to particles formed by complementation between supernatant extracts of chl A and chl B mutants. There are three kinds of particles of different buoyant densities (1.10, 1.18 and 1.23); ATPase binds only to the 1.10 and 1.18 particles. The kinetics of incorporation have been studied. ATPase begins to be incorporated into the 1.10 particles after 10 min of incubation up to a maximum at 20 min: from 30 min, ATPase is incorporated only into 1.18 particles and the amount of incorporated ATPase increased in proportion with the peak of 1.18 particles. These kinetics have a hyperbolic pattern. In order to explain the mechanism of assembly involved in complementation, two hypotheses are proposed.


Ca-2+-stimulated membrane phosphorylation and ATPase activity of the human erythrocyte. 1. Human erythrocyte membranes were preincubated with ethyleneglycolbis-(beta-aminoethyl)-N,N' tetraacetate (EGTA) and subsequently labelled for short periods with micromolar concentrations of [8-3-H, gamma-32-P]ATP. Under these conditions, and at temperatures smaller than or equal to 22 degrees C, both ATP hydrolysis and membrane phosphorylation were stimulated by Ca-2+. 2. The properties of the Ca-2+-stimulated ATP hydrolysis and associated phosphorylation of a 150 000 molecular weight protein component, previously described (Knauf, P. A., Proverbio, F. and Hoffman, J. F. (1974) J. Gen. Physiol. 63, 324-336), have been studied. The behavior of the phosphorylated component, ECaP, has properties consistent with its role as a phosphorylated intermediate of Ca-2+-ATPase activity, including: (1) similar dependence of the steady-state level of ECaP and Ca-2+-ATPase on ATP concentration; (2) rapid turnover apparent upon the addition of excess non-radioactive ATP; and (3) good correlation between the steady-state levels of Ca-2+-dependent phosphorylation and Ca-2+-ATPase activity in separate preparations possessing variable specific activity. Addition of excess EGTA to ECaP caused only partial dephosphorylation. Sensitivity of Ca-2+-stimulated ATP hydrolysis and associated phosphorylation to micromolar concentrations of Ca-2+ implicates this activity in the "high-affinity" Ca-2+-pump system of the human erythrocyte (Schatzmann, H. J. (1973) J. Physiol. London 235, 551-569).


Purification of cardiac myosin. Application to hypertrophied myocardium. A method is described for the preparation of high purity myosin from small amounts of cardiac muscle. The method employs homogenization and prolonged extraction of the cardiac tissue. Purification is achieved through three successive precipitation-dissolution cycles and without the use of column chromatographic techniques. Purity of the myosin preparation is assessed at various stages of the purification procedure by sodium dodecylsulfate-acrylamide gel electrophoresis and by measurement of RNA and nucleoprotein content. With 1.5-2.0 g of rabbit right ventricle as the starting tissue, this method yields 4-6 mg myosin per g wet tissue. The method is also shown to give similar results with rabbit right ventricles hypertrophied by pulmonary stenosis.


Differences in the binding to fibrin of native plasminogen and plasminogen modified by proteolytic degradation. Influence of omega-aminocarboxylic acids. Pretreatment of native plasminogen with plasmin or activators resulted in a pronounced increase in the binding of plasminogen to fibrin. The pretreated plasminogen was considered to be identical to the proteolytically degraded proenzyme with NH2-terminal lysine, valine or methionine, which is formed as an intermediate stage during activation of plasminogen. Bound plasminogen could be extracted by 6-aminohexanoic acid indicating a reversible binding between plasminogen and fibrin. Adsorption of pretreated plasminogen decreased when increasing concentrations of 6-aminohexanoic acid or trans-4-aminomethylcyclohexane-1-carboxylic acid (t-AMCHA) were present during fibrin formation. The concentration of amino acid producing a decrease in the binding of pretreated plasminogen to 0.5 of the amount bound in the absence of amino acid was 8.0-10(-5) M with 6-aminohexanoic acid and 1.7.10-5 M with t-AMCHA. The decrease in binding is most likely related to an effect of the amino acids on plasminogen, since agarose gel electrophoresis of pretreated plasminogen in the presence of 6-aminohexanoic acid or t-AMCHA showed a cathodic shift in mobility at the same range of concentrations of amino acid, which produced the decrease in binding of plasminogen to fibrin. Evidence is provided that the decrease in binding of proteolytically degraded plasminogen may result in an inhibition of fibrinolysis caused by activators.


Preparation of prostatic plasma membranes Distribution of (Na-+, K-+)-ATPase and Mg-2+-ATPase in the rat ventral prostate. 1. A method is described for the isolation of a plasma membrane fraction from rat ventral prostate. This fraction is greatly enriched in (Na-", K-+)-ATPase but also contains a small amount of Mg-2+-ATPase and 5'-nucleotidase activities. 2. The activity ratio (Mg-2+ plus Na-+ plus K-+/Mg-2+ or Mg-+ plus Na-+) of the plasma membrane (Na-+, K-+)-ATPase was 1.7. The enzyme system specifically requires ATP as substrate and is inhibited by Ca-2+ or ouabain.


Polar head-group and acyl side-chain requirements for phospholipid-dependent (Na-+ plus K-+)-ATPase. The abilities of different phospholipids to reactivate a lipid-depleted (Na-+ plus K-+)-ATPase (EC 3.6.1.3) have been compared. The phospholipids contained either the same group of hydrocarbon chains withhh different polar groups, or different hydrocarbon groups wit the same polar group; they were prepared by enzymic modification of the polar group of phosphatidylserine isolated from bovine brain and of phosphatidylcholine extracted from egg. Only the acidic phospholipids reactivated the ATPase but the amount of reactivation depended on the nature of the hydrocarbon chains as well as the polar group. These findings are discussed in relations to the compositions of the two different groups of fatty acyl chains constituting the hydrocarbon portions of the phospholipids.


Effect of carbonylcyanide m-chlorophenylhydrazone on the calcium-stimulated ATPase activity of erythrocyteghosts. Incubation of etythrocyte ghosts with carbonylcyanide m-chlorophenyl-hydrazone (CCCP) plus Ca-2+ resulted in inactivation of the Ca-2+ -stimulated ATPase activity. Omission of Ca-2+ or lowering of the temperature below 25 degrees C eliminated the inhibitory effect, as also did the presence of ATP during the incubation. On the other hand, the addition of beta-mercaptoethanol did not influence the Ca-2+ -dependent inhibition by CCCP. Compared with the level of CCCP which uncouples oxidative phosphorylation, a rather high level (0.5 mM) of CCCP was required to inhibit the ATPase activity in ghosts. However, once the inhibition had been accomplished, almost all of the CCCP could be removed from the ghost membrane by washing with a Ca-2+ -containing solution, without affecting the inhibition of ATPase. If ethylene-glycol-bis(beta-aminoethyl ether)-N,N'-tetraacetic acid was included in the washing medium, the inhibition of ATPase was nearly completely reversed by washing. The results indicate that only the Ca-2+ -stimulated, Mg-2+ -ATPase was inhibited by 0.5 mM CCCP, while the remaining components of the ATPase activity were slightly inhibited by higher levels of the uncoupler. Low levels of CCCP (0.1 mM) stimulated the Mg-2+ -ATPase slightly. CCCP was much more specific for the Ca-2+ -stimulated ATPases than N-(1-naphthyl)maleimide, an unusually effective sulfhydryl reagent, and the requirement of Ca-2+ for inactivation was also quite specific.


Evidence for a plasma membrane calcium pump in bovine adrenal medulla but not adrenal cortex. Continuous sucrose density gradient subfractions from bovine adrenal medullary microsomes were found to accumulate 45-Ca-2+ in the presence of ATP and ammonium oxalate mainly in subfractions of intermediate density. (Na-++K-+)-ATPase (plasma membrane marker) and Ca-2+-ATPase activities were also concentrated in these intermediate subfractions but thiamine pyrophosphatase (Golgi apparatus marker) was not. NADH oxidase (endoplasmic reticulum marker) activity was distributed throughout all subfractions. 45-Ca-2+ accumulation in adrenal cortical microsomes was found to rise and fall in parallel with thiamine pyrophosphatase but not with (Na-++K-+)-ATPase or NADH oxidase activities. Accumulation of 45-Ca-2+ in membrane vesicles in these experiments suggests the existence of a calcium transfer mechanism in plasma membranes of the adrenal medulla but not adrenal cortex.


ATP-dependent calcium accumulation in brain microsomes. Enhancement by phosphate and oxalate. 1. ATP-dependent calcium uptake by a rabbit brain vesicular fraction (microsomes) was studied in the presence of phosphate or oxalate. These anions, which are known to form insoluble calcium salts, increased the rate of calcium uptake and the capacity of the vesicles for calcium accumulation. 2. The degree of activation depended on the concentration of phosphate or oxalate. Under optimal conditions, phosphate promoted a 5-fold increase in the amount of calcium stored at steady state. This level was 200-250 nmol Ca-2+/mg protein. 3. Initial rate of calcium uptake followed Michaelis-Menten kinetics with an apparent Km for calcium of 6.7-10-minus 5 M and a V of 44 nmol/min per mg protein. Optimal pH was 7.0. With 2 mM ATP, optimal Mg-2+ concentration was 2 mM. 4. Dintrophenol and NaN3 inhibited calcium uptake in a mitochondria-enriched fraction but not in the microsomal fraction. 5. Calcium uptake activity was compared in the six subfractions prepared from the whole microsomal fraction by means of a sucrose density gradient fractionation. 6. The Mg-2+-dependent ATPase activity of brain microsomes was activated by calcium. Maximal activation was attained with 100 muM CaCl2. Greater calcium concentrations caused a progressive inhibition. 7. The data suggest that the ATP-dependent calcium uptake in brain microsomes, as in muscle microsomes, is brought about by an active transport process, calcium being accumulated as a free ion inside the vesicles.


Ouabain-insensitive Na+ stimulation of an Mg-2+ -dependent ATPase in kidney tissue. 1. Freshly prepared microsomal fractions of the outermost cortex of guinea pig kidney show an Mg-2+-dependent ATPase activity which is partially inhibited by 100 mM NaCl, LiCl, KCl, RbCl, CsCl, NH4Cl or choline chloride. 2. If the microsomal preparation is aged by storage at 4 degrees C for 10-15 days, the Mg-2+-dependent activity shows stimulation by Na-+  and Li-+ but not by K-+, Rb-+, Cs-+, NH4-+ or choline. 3. Stimulation is similar with sodium salts of Cl-minus, HCO3-minus, CH3COO-minus, BR-minus, SO4-2-minus or methylsulphonate. 4. Stimulation is insensitive to 1 mM and 10 mM ouabain. 5. Stimulation is unaltered by the presence of 0.5 mM ethyleneglycol-bis-(beta-aminoethyl ether)N,N'-tetracetic acid. 6. Stimulation is 100% inhibited by 2 mM ethacrynic acid, a concentration which inhibits only 30% of the Mg-2+-dependent ATPase and 50% of the (Na-++K-+)-stimulated ATPase. 7. Some of these characteristics coincide with those of an ouabain-resistant, K-+-independent, ethacrynic acid-sensitive mode of Na-+ extrusion out of guinea pig kidney cortex cells.


Inactivation of (Na-++K-+)-stimulated ATPase by a cytotoxic protein from cobra venom in relation to its lytic effects on cells. The mechanism of action of the cytotoxic protein P6 isolated from cobra venom (Naja naja) which shows preferential cytotoxicity particularly to Yoshida sarcoma cells has been studied by its effects on the membrane-bound enzyme (Na-++K-+)-ATPase (ATP phosphohydrolase, EC 3.6.1.3) of a variety of cell systems. Evidence obtained with Yoshida sarcoma cells, dog and human erythrocytes and three tissue culture cell lines KB (human oral carcinoma), Hela (human cervix carcinoma) and L-132 (human lung embryonic) shows that inhibition of (Na-++K-+)-ATPase by the P6 protein can be correlated with its lytic activity. (Na-++k-+)-ATPase of Yoshida sarcoma membrane fragments inactivated by P6 protein could be reconstituted by the addition of phosphatidylserine and phosphatidic acid. It is conceivable that lysis of cells by the P6 protein may be due to an imbalance of K-+ and Na-+ in the cell which leads to swelling and disintegration of the membrane structure. Observations indicate that the P6 protein combines with membrane constituents of susceptible cells. The overall evidence suggests that both the specificity of its protein structure and the highly basic nature of the P6 protein are factors which enable it to compete with the lipid moiety maintaining the (Na-++k-+)-ATPase of the susceptible cells in proper conformation for activity.


Hormone responsiveness of plasma membrane-bound enzymes in normal and regenerating rat liver. The hormonal responsiveness of plasma membrane-bound enzymes (Na-+-K-+)-ATPase and adenylate cyclase has been investigated in normal and regenerating rat liver. (Na-+-K-+)-ATPase basal activity is not affected by surgery and only slightly affected by partial hepatectomy; its response to epinephrine and cyclic AMP is decreased only 15 h after hepatectomy. Adenylate cyclase activity of plasma membranes from untreated animals is stimulated by parathyroid hormone and thyroxine; partial hepatectomy increased basal activity as well as the stimulation exerted by the aforementioned hormones, when glucagon and epinephrine sensitivity is essentially unaltered.


Discrepancies in the measurement of changes in blood flow using flow- and velocity-sensitive electromagnetic probes. Blood flow was measured in the descending thoracic aorta of the anaesthetised dog using a perivascular flow probe and electromagnetic flowmeter. Blood velocity was measured in the same vessel with an intravascular velocity probe and electromagnetic flowmeter. Changes in blood pressure, flow and velocity, were induced by intravenous administration of noradrenaline, angiotensin, isoprenaline and aminophylline. If the velocity probe was within the cuff of the flow probe, percentage changes in flow and velocity were comparable, but if the velocity probe was "upstream" or "downstream" from the cuff, discrepancies between percentage drug-induced changes in flow and velocity were of the order of 30% and, in 12 out of 80 observations, in opposite directions. Measuring absolute flow with the velocity-sensitive device produced variable results showing a systematic underestimate of flow estimated from velocity, compared with flow measured with the perivascular probe. It is concluded that the measurement of blood flow and changes in blood flow, with the intravascular velocity probe, is liable to produce inaccurate data.


[Study of the stationary kinetics of the myosin ATPase reaction in the presence of calcium]. Calcium activation of myosine ATPase is investigated. Dependency of the hydrolysis rate on total concentration of metal ions and substrate and kinetics of the reaction under [ATP]tot. equals [Ca]tot. are studied. Dependency of upsilon on [Ca]tot, was found to be complex under comparable concentrations of ATP and calcium at the range of 0.1-1 mM. The data obtained show that the complex character of myosine ATPase reaction in the presence of calcium is due to a dual nature of the enzyme activation with calcium ions: CA+2-induced transition of myosine from low-active into active form and the formation of new easily hydrolyzible substrate (Ca ATP). Low-active form of the enzyme is a protein which is tightly bound with magnesium while active myosine is free of bivalent cations or is weakly bound with calcium. This two enzyme forms are indistinguishable kinetically. It is shown that Ca+2 at high concentrations is capable to substitute magnesium ions bound to the protein molecule. Activating effect of free metal on myosine ATPase is found to come to the protection of the enzyme from magnesium inhibition.


[Alkylating compounds, inhibitors of mitochondrial ATP-synthetase]. The esters of aromatic acids with two electrophilic groups were found to inhibit mitochondrial ATP-synthetase like oligomycin. The substances without electrophilic sites or with one electrophilic groups have no the oligomycin-like effect on mitochondria. Aromatic acids with two alkylating groups are also inefficient.


Cardiotonic steroids: correlation of sodium-potassium adenosine triphosphate inhibition and ion transport in vitro with inotropic activity and toxicity in dogs. 1. A new series of cardiotonics based on five steroid nuclei has been evaluated for inhibition of Na-+/K-+-ATPase and Rb uptake by red blood cells, and for inotropic activity and toxicity in dogs. Structure-activity relationships are discussed. 2. The in vitro tests can be used satisfactorily to predict inotropic activity, but not toxicity or therapeutic ratio. 3. Although compounds with greatly improved therapeutic ratios relative to ouabain and tolusin have been obtained, they proved to be strongly emetic in the conscious dog.


Extended deep femoral angioplasty: an alternative to femoropopliteal bypass. The results of extended deep femoral angioplasty (EDFA) have been alaysed for 74 legs in 72 patients. In 69 per cent a femoropopliteal bypass would have been possible, so DEFA is considered here as an alternative operation. The success rate was 66-9 per cent, the failure rate 25-6 per cent and no effect was achieved in 7-5 per cent. Diabetes adversely affected the results. Of the failures, 45 per cent were diabetic, while the incidence of diabetes was only 14 per cent in the group where the operation was successful. The long term results of the operation were reasonable, 75 per cent being successful for 10-39 months. Age had no bearing on success or failure. The effects of EDFA were most dramatic on intermittent claudication. In 52 per cent it was abolished and in 92 per cent claudication distance was increased to over 200 yards. Good results were achieved after failed lumbar sympathectomy and failed femoropopliteal bypass. Successful results of EDFA could be predicted best by inspection of angiograms. Success or failure could have been predicted from the state of the run-off in 81 per cent of the cases. Claudication and rest pain were the most relieved by EDFA, followed by isachaemic ulceration of the leg. Gangrene of digits was helped but less than had been hoped. We conclude that the indications for femoropopliteal bypass are limited to cases of digital gangrene where angiography shows that bypass is possible or where an EDFA operation has failed.


Pre-eclampsia in pregnancies by different fathers: immunological studies. Immunological studies were performed on a woman with severe pre-eclamptic toxaemia in a second pregnancy. This pregnancy followed a normal twin pregnancy by a different father four years earlier. Both fathers were also studied. In the mixed lymphocyte culture the patient's lymphocytes reacted eight times as strongly against father 2's cells as against those from father 1. If studies along these lines are performed when a women has toxaemic and non-toxaemic pregnancies by different fathers information may be obtained on immunogenetic aetiological factors which may be of more value than that derived from the study of large unselected populations.


Treatment of gonorrhoea in the female with fortified procaine penicillin plus probenecid and with spectinomycin. 513 cases of uncomplicated gonorrhoea in females with positive results to cultures were treated with two regimens of treatment between January 1, 1972, and February 28, 1973, at the Department for Females, Bangrak Hospital, Bangkok. The highest incidence was found in the age group 20 to 24 years (215 out of 513). The patients were divided into two groups. In the first, probenecid 1 g. was given orally 1 hour before an intramuscular injection of 4 million units fortified procaine penicillin, and in the second an intramuscular injection of 2 g. spectinomycin dihydrochloride was given. 335 cases were followed in the first group and there were no failures; 132 cases were followed in the second group and there was one failure (0.7 per cent.). The fact that gonococci were found in the rectum in 23.7 per cent. of cases (117 out of 493) indicates that rectal tests are of considerable importance and should be included in the routine investigations. There was no evidence that gonorrhoeal infection of the rectum was more resistant to treatment than infection at other sites.


A comparison of methods of evaluating myocardial contractility. An investigation was carried out in dogs to determine how the acceleration of blood in the aorta (dV/dt), as a new index of myocardial contractility compared with existing indices and how they correlated with each other. It was found that the indices derived from velocity and flow of blood, dV/dt max and dQ/dt max, and from intraventricular pressure, dP/dt max, correlate well with each other but there is less agreement between them and the reciprocal of the Pre-Ejection Period, l/PEP. The ratio dP/dt max divided by I.P. correlated well with dQ/dt max divided by I.Q. and dQ/dt max divided by Q. max but not so well with dV/dt max divided by I.V. (instantaneous velocity) of dV/dt max divided by V. max and the ratios dQ/DT DIVIDED BY I.Q. and dQ/dt divided by Q max as well as l/PEP. In view of the lack of agreement of quantitative definition of myocardial contractility, the ratio dV/dt divided by V. max would have several practical advantages as an indicator of the inotropic state of the heart; these are that the probe used to establish descending aortic blood velocity does not require calibration, and the signal can be obtained by a relatively non-invasive technique that is suitable for patient care and yet agrees with other established indices of myocardial contractility.


Identification and turnover of glycosaminoglycans in rat kidneys. The turnover of sulfate label in crude glycosaminoglycan fractions from rat kidney cortex, medulla, and papilla has been determined. Heparan sulfate, chondroitin sulfate, dermatan sulfate, and hyaluronate have been separated electrophoretically and their specific activities determined after injection of labeled sulfate or glucose. The half-lives of the sulfated glycosaminoglycans are within the ranges found for other organs and tissues, but hyaluornate has a somewhat faster turnover in the kidney than elsewhere.


Studies on the control of Mycoplasma gallisepticum in hatching eggs. Lincomycin-spectinomycin, while effective in eliminating Mycoplasma gallisepticum from hatching eggs when administered via the yolk sac, had a marked deleterious effect on hatch-ability when given by this route. On the other hand when lincomycin-spectinomycin was injected into the air cell there was a reduction in the level of experimental infection with an acceptable effect on hatchability.


Screening for hypertension in a high school population. Of 15 594 high school students (ages, 15 to 20) whose blood pressure was measured in a screening program, 350 (2.2%) has hypertensive readings (150 mm Hg or more systolic, or 95 mm Hg or more diastolic, or both). The mean blood pressure for the boys was 125.0  plus or minus 12.1/71.8 plus or minus 10.9 mm Hg, and for the girls, 119.8 plus or minus 10.2/72.3 plus or minus 9.2 mm Hg. The parents of the students with hypertensive readings were advised to send their children to a physician. By 6 months, of the 232 who were followed up, 156 (67.2%) has visited a physician and in 19 cases (12.2%) the physician had confirmed the hypertensive readings. Only one student, an asymptomatic 17-year-old boy whose hypertension had not previously been detected, was found to have secondary hypertension, which was relieved surgically. Of the 18 hypertensive students 4 are currently receiving antihypertensive medication and 8 continue to have their blood pressure monitored. The mean blood pressures recorded in the physicians' offices averaged 23.7/11.1 mm Hg less than those recorded in the schools. One reason for this was that none of the physicians used pediatric cuffs, but these were required by 62.4% of the students at the screening. Hence, the intravascular blood pressure was probably underestimated in a number of cases in the physicians' offices.


Increased urinary excretion of nucleic acid and nicotinamide derivatives by rats after treatment with alkylating agents. Rats treated with di(2-chloroethyl)methylamine (HN2), N-methyl-N-nitrosourea (MNUA) and N-ethyl-N-nitrosourea (ENUA) excrete significantly larger amounts of deoxycytidine (dC) and thymidine in their urine 0-24 h after treatment. Ethyl methanesulphonate (EMS) and dimethylnitrosamine (DMN) gave negative results in this respect but all five alkylating agents increased the excretion of 1-methyl-nicotinamide (1-meNmd). In addition, a larger quantity of 7-methylguanine (7MG) and uric acid was excreted after DMN treatment. 1,4-Dimethanesulphonoxybutane (myleran), 2,2-dichlorovinyl dimethyl phosphate (dichlorvos), 5-fluorouracil (5FU), cytosine arabinoside (araC), 2-acetylaminofluorene (AAF) and 7-bromomethylbenz-[a]anthracene (7-BrMBA) gave negative results.


Inhibitory effect of hydroxyurea on [3-H]leucine incorporation in human peripheral lymphocytes. Hydroxyurea (HU), generally considered to be a specific inhibiter of DNA synthesis, has an inhibitory effect on the incorporation of TCA-precipitable [3-H]leucine in peripheral lymphocytes. This action is not secondary to the inhibition of DNA synthesis since incorporation of [3-H]leucine is unaffected when DNA synthesis is inhibited by 5-fluorodeoxyuridine (FUdR); it does not appear to be directly related to inhibition of RNA synthesis; and it is not mediated at the level of translation since HU has no effect on protein synthesis in rabbit reticulocytes. The relevance of these findings to the use of HU as a DNA inhibitor is discussed.


Acute myocardial infarction in toxic cardiomyopathy without coronary obstruction. Confluent left ventricular scar without significant coronary obstruction has been found in alcoholic subjects at autopsy. To evaluate the pathogenesis, 12 patients with chronic alcoholism and severe precordial pain persisting 4-24 hours were observed clinically. Cardiac isoenzymes of lactic dehydrogenase rose in serum. ST segment was elevated in anterior or posterior ECG leads, and abnormal Q waves appeared. Hypertnesion and hypercholesterolemia were present in two (group B) but not in the ten (group A). The latter exhibited no significant obstructive disease, based on coronary angiography in seven survivors and postmortem examination of the remaining three. Clinical evidence, as well as the quantitative assessment of platelets, made arterial thromboembolism an unlikely cause for the symptoms. Neither hemotologic or systemic disease affecting myocardium was present. The morphology of the left ventricle in three autopsies was compared with that of patients with alcoholism who had no cardiac disease, cardiomyopathy, or an asymptomatic scar. All had accumulation of Alcian positive glycoprotein in the interstitium. The patients with cardiac disease also had interstitial fibrosis which was characterized, particularly in the acute infarction group, by concentric periarterial fibrosis. Restriction of coronary vasodilation by this process during periods of high blood flow requirements was postulated as a basis for infarction.


A simplified method for quantitating left-to-right shunts from arterial dilution curves. This report presents a new, simple and reliable method of quantitating left-to-right shunts from arterial dilution surves. When indicatior (e,g., indocyanine green) is injected centrally (right venticle or pulmonary artery) and blood is withdrawn rapidly from either the aorta or other central systemic artery, in the presence of a left-to-right shunt to recorded indicator-dilution curve usually shows two discrete peaks: p-1 of the magnitude of the shunt expressed as percent of pulmonary blood flow, Experimental and clinical data demonstrate that this method is accurate in quantitating shunts from such curves.


Molecular size difference of urinary heparan sulfates from normal individuals and genetic mucopolysaccharidoses. Heparan sulfates were isolated from the urine of normal individuals and patients with genetic mucopolysaccharidoses after exhaustive digestion with chondroitinase ABC. Electrophoresis of these preparations on cellulose acetate membrane revealed one spot corresponding in mobility to reference heparan sulphate in barium acetate buffer, while electrophoresis in 0.1 M HCl resulted in two distinct spots for each case; one corresponded in migration rate to reference heparan sulfate, and the other was faster in mobility than reference heparan sulfate but slightly retarded when compared with reference heparin. On thin-layer gel filtration on Sephadex G-200 (superfine) heparan sulfate from normal urine was polydispersed in character and its molecular size was larger than those of other preparations. Heparan sulfates from Hunter's and Sanfilippo's urine were monodispersed and small in molecular size. The molecular size of heparan sulfate from Sanfilippo's urine was the smallest of all. Heparin sulfate from Hurler's urine appeared to be composed of two populations; one corresponded in molecular size to heparan sulfate from normal urine, and the other corresponded to that of Hunter's urine.


Renal lesions accompanying poison oak dermatitis. Renal damage associated with poison oak dermatitis is extremely rare in humans after exposure to urushiol antigen. Three renal lesions have been described: proliferative glomerulonephritis, arteritis, and membranous nephropathy. The present study reports on three patients who developed nephropathy after exposure to poison oak. One patient was studied by renal biopsy (including electron microscopy and immunofluorescence techniques) and another by autopsy findings. One of these patients had a typical membranous nephropathy, the other, proliferative glomerulonephritis with necrotizing arteritis and glomerulitis. In the patient with membranous nephropathy antibody to urushiol was discovered by passive cutaneous anaphylaxis in the guinea pig.